LOCUS       pTwist_Lenti_CAG        9121 bp    DNA     circular SYN 11-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9121)
  AUTHORS   siwong
  TITLE     Direct Submission
  JOURNAL   Exported Dec 5, 2024 from SnapGene 8.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..658
                     /label=chicken beta-actin promoter
     intron          792..1809
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     misc_feature    1853^1854
                     /label=Your gene here
     misc_feature    1868..2456
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2339..2350)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2528..2761
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2839..2960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3000..3135
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3156..3174)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3184..3200)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3342..3797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3823..3927
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3928..3996,3997..4788)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4959..5547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5835..5856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5871..5876,5877..5894,5895..5901)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5909..5925
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5933..5949
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5970..5988
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6016..6242
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6243..6423
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6470..6595
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7088..7321
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7506..7550
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        7725..8225
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             8247..8846
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             8247..8846
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    8888..9005
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      421 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      481 gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg
      541 cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      601 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcgcg
      661 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
      721 acaccgggac cgatccagcc tcccctcgaa gcttacatgt ggtaccgagc tcggatcctg
      781 agaacttcag gggagtcgct gcgacgctgc cttcgccccg tgccccgctc cgccgccgcc
      841 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      901 ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg
      961 tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg
     1021 gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc
     1081 ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg
     1141 agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc
     1201 gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa
     1261 ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc
     1321 cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg
     1381 ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc
     1441 cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg
     1501 tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg
     1561 cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa
     1621 atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct
     1681 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     1741 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
     1801 ttcctacagc tcctgggcaa cgtgctggtc tgtgtgctgg cccatcactt tggggccggc
     1861 cagatctaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1921 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1981 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2041 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2101 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2161 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2221 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     2281 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     2341 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2401 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg
     2461 actttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     2521 gggggactgg aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact
     2581 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     2641 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     2701 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     2761 agtacgtata gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     2821 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2881 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2941 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca
     3001 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3061 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3121 gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt attacgcgcg
     3181 ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
     3241 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     3301 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     3361 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3421 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3481 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3541 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3601 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3661 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3721 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3781 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3841 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3901 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3961 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4021 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4081 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4141 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4201 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4261 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4321 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4381 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4441 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4501 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4561 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4621 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4681 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4741 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4801 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4861 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4921 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4981 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5041 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5101 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5161 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5221 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5281 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5341 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5401 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5461 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5521 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5581 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5641 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5701 agcgagtcag tgagcgagga agcggtagag cgcccaatac gcaaaccgcc tctccccgcg
     5761 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5821 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     5881 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     5941 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     6001 gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga
     6061 tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag
     6121 taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac
     6181 gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa
     6241 acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6301 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6361 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6421 gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg
     6481 caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac
     6541 gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat
     6601 taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa
     6661 aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa
     6721 tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc
     6781 ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg
     6841 tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga tagaggatga
     6901 gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag
     6961 gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac
     7021 cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag
     7081 tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag
     7141 cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga
     7201 acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca
     7261 tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc
     7321 tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta
     7381 gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca
     7441 gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc
     7501 aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt
     7561 ttaacataac aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg
     7621 taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt
     7681 caccattatc gtttcagacc cacctcccaa ccccgagggg acccttgcgc cttttccaag
     7741 gcagccctgg gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg
     7801 gcgccgaccc tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg
     7861 ccgctaccct tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt
     7921 ccttgcggtt cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc
     7981 tcgcagacgg acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca
     8041 atagcggctg ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg
     8101 cggggtgtgg ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca
     8161 agcctccgga gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc
     8221 cccagggggt accaccggag cttaccatga ccgagtacaa gcccacggtg cgcctcgcca
     8281 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     8341 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     8401 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     8461 ccgtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     8521 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     8581 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     8641 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     8701 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     8761 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     8821 gcatgacccg caagcccggt gcctgagaat tctagatctt gagacaaatg gcagtattca
     8881 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     8941 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     9001 attttcgggt ttattacagg gacagcagag atccactttg gcgccggctc gagggggccc
     9061 ggggttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     9121 g
//
