LOCUS       pTwist_Lenti_SFF        7718 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 7718)
  AUTHORS   siwong
  TITLE     Direct Submission
  JOURNAL   Exported Dec 5, 2024 from SnapGene 8.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..408
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    427^428
                     /label=Your gene here
     misc_feature    465..1053
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(936..947)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1125..1358
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1436..1557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1597..1732
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(1753..1771)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1781..1797)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1939..2394
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2420..2524
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2525..2593,2594..3385)
                     /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      3556..4144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4432..4453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4468..4473,4474..4491,4492..4498)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4506..4522
                     /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     4530..4546
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4567..4585
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4613..4839
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4840..5020
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5067..5192
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5685..5918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6103..6147
                     /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        6322..6822
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             6844..7443
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             6844..7443
                     /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    7485..7602
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gtaacgccat tttgcaaggc atggaaaaat accaaaccaa gaatagagaa gttcagatca
       61 agggcgggta catgaaaata gctaacgttg ggccaaacag gatatctgcg gtgagcagtt
      121 tcggccccgg cccggggcca agaacagatg gtcaccgcag tttcggcccc ggcccgaggc
      181 caagaacaga tggtccccag atatggccca accctcagca gtttcttaag acccatcaga
      241 tgtttccagg ctcccccaag gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc
      301 agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa aagagctcac
      361 aacccctcac tcggcgcgcc agtcctccga cagactgagt cgcccgggga gacccaagct
      421 ggctagctct agagggcccg tttaaacccg ctgatcagcc tcgaaatcaa cctctggatt
      481 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
      541 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
      601 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
      661 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
      721 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
      781 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
      841 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
      901 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
      961 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     1021 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa
     1081 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     1141 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     1201 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     1261 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     1321 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca
     1381 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     1441 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     1501 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     1561 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     1621 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     1681 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg
     1741 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     1801 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1861 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1921 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1981 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2041 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2101 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2161 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2221 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2281 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2341 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     2401 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2461 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2521 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2581 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2641 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2701 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     2761 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2821 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2881 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     2941 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3001 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3061 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3121 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3181 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3241 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3301 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3361 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     3421 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     3481 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3541 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3601 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3661 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3721 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3781 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3841 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3901 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3961 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4021 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4081 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4141 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4201 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4261 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4321 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     4381 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     4441 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     4501 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     4561 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct
     4621 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa
     4681 ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta
     4741 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc
     4801 agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag
     4861 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     4921 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     4981 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
     5041 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
     5101 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
     5161 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     5221 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     5281 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     5341 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     5401 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     5461 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     5521 cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     5581 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     5641 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     5701 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     5761 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     5821 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     5881 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     5941 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     6001 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     6061 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     6121 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     6181 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     6241 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     6301 ctcccaaccc cgaggggacc cttgcgcctt ttccaaggca gccctgggtt tgcgcaggga
     6361 cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca
     6421 ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt gggccccccg
     6481 gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg
     6541 acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca gcgccaggga
     6601 gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc agcagggcgc
     6661 gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg
     6721 gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg cacgtcggca
     6781 gtcggctccc tcgttgaccg aatcaccgac ctctctcccc agggggtacc accggagctt
     6841 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     6901 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     6961 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     7021 gacatcggca aggtgtgggt cgcggacgac ggcgccgccg tggcggtctg gaccacgccg
     7081 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     7141 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     7201 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     7261 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     7321 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     7381 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     7441 tgagaattct agatcttgag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     7501 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     7561 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     7621 agcagagatc cactttggcg ccggctcgag ggggcccggg gttattaata gtaatcaatt
     7681 acggggtcat tagttcatag cccatatatg gagttccg
//
