LOCUS       pTwist_Lenti_CMV        7727 bp DNA     circular SYN 22-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7727)
  AUTHORS   siwong
  TITLE     Direct Submission
  JOURNAL   Exported Oct 28, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7727
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    603^604
                     /label=Your gene here
     misc_feature    670..1258
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(1141..1152)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1330..1563
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1641..1762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1802..1937
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(1958..1976)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1986..2002)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2144..2599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2625..2729
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2730..2798,2799..3590)
                     /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      3761..4349
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4637..4658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4673..4678,4679..4696,4697..4703)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4711..4727
                     /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     4735..4751
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4772..4790
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4818..5044
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5045..5225
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5272..5397
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5890..6123
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6308..6352
                     /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        6527..7027
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             7039..7434
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7494..7611
                     /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 gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctgagacc caagctggct
      601 agcgatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa acccgctgat
      661 cagcctcgaa atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
      721 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
      781 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
      841 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
      901 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
      961 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     1021 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     1081 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     1141 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     1201 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt
     1261 cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     1321 aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta
     1381 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     1441 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     1501 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     1561 gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     1621 gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca
     1681 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1741 ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc
     1801 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     1861 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     1921 aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg
     1981 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     2041 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     2101 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     2161 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2221 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2281 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2341 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2401 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2461 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2521 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2581 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2641 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2701 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2761 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2821 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2881 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2941 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3001 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3061 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3121 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3181 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3241 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3301 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3361 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3421 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3481 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3541 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3601 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3661 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3721 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3781 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3841 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3901 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3961 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4021 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4081 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4141 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4201 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4261 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4321 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4381 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4441 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4501 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     4561 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     4621 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     4681 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     4741 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     4801 tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac
     4861 gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga
     4921 agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg
     4981 acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat
     5041 aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     5101 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     5161 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     5221 tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga
     5281 cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt
     5341 acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt
     5401 attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag
     5461 aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt
     5521 aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca
     5581 tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc aaccctctat
     5641 tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa
     5701 gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg
     5761 aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga
     5821 accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc
     5881 agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc
     5941 agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca
     6001 gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg
     6061 catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct
     6121 cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc
     6181 tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga
     6241 cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca
     6301 gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg
     6361 gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt
     6421 ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata
     6481 ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccttgc gccttttcca
     6541 aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag
     6601 cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt
     6661 cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg
     6721 ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac
     6781 cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc
     6841 caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga
     6901 ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg
     6961 caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc
     7021 tccccagggg gtaccaccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa
     7081 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     7141 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     7201 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     7261 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     7321 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     7381 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggcctgcag
     7441 ctgcagtagt aagaattcta gatcttgaga caaatggcag tattcatcca caattttaaa
     7501 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     7561 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat
     7621 tacagggaca gcagagatcc actttggcgc cggctcgagg gggcccgggg ttattaatag
     7681 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
