LOCUS       AAV01-02_pAAV-TF        5762 bp    DNA     circular SYN 06-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5762)
  AUTHORS   Demo User
  TITLE     Direct Submission
  JOURNAL   Exported Oct 6, 2025 from SnapGene 8.1.1
            https://www.snapgene.com
COMMENT     http://www.invitrogen.com/
FEATURES             Location/Qualifiers
     source          1..5762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=5'-ITR
     enhancer        282..585
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1020..1152
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1190..1199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1196..1198,1199..1201,1202..1915)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1972..2513
                     /label=WPRE
     CDS             complement(2443..2454)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2524..2547
                     /label=BGH ddPCR F1-adapted
     polyA_signal    2527..2734
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2535..2557
                     /label=BGH_F
     primer_bind     2564..2583
                     /label=BGH ddPCR probe
     primer_bind     2565..2585
                     /label=BGH_probe
     primer_bind     complement(2587..2604)
                     /label=BGH_R
     primer_bind     complement(2595..2618)
                     /label=BGH ddPCR R1-adapted
     repeat_region   2822..2951
                     /label=3'-ITR
     rep_origin      3128..3583
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3610..3714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3715..3783,3784..4575)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVVMVTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4186..4204
                     /label=AMP F
     primer_bind     4342..4361
                     /label=AMP ddPCR F
     primer_bind     4365..4384
                     /label=AMP ssPCR probe
     primer_bind     complement(4402..4421)
                     /label=AMP ddPCR R
     rep_origin      4746..5334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      181 agccatgctc taggaagatc ggaattcgcc cttaagctag atagttatta atagtaatca
      241 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      301 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      361 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      421 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      481 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      541 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggcacc tgcagagggc
      601 cctgcgtatg agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc
      661 tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc
      721 ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt
      781 cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca
      841 ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg
      901 gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg
      961 ggcacgggcg cgaccatctg cgctgcggcc gcactagttg gtcgtgaggc actgggcagg
     1021 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     1081 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     1141 ttctctccac aggtgtccag gcggccgctc tagaggatcc ccaccggtcg ccaccatggt
     1201 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     1261 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     1321 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     1381 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1441 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1501 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1561 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1621 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1681 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1741 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1801 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1861 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     1921 ccgcgactct agctcgagca ctagtccccg ggaccgcggc acgcgtgatc caatcaacct
     1981 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2041 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2101 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2161 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2221 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     2281 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     2341 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     2401 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     2461 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgagatctg
     2521 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2581 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2641 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2701 aggattggga agacaatagc aggcatgctg gggactcgag ttaagggcga attcccgata
     2761 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     2821 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     2881 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     2941 cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg tcgtgactgg
     3001 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     3061 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     3121 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3181 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3241 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3301 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3361 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3421 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3481 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3541 aaatttaacg cgaattttaa caaaatatta acgtttataa tttcaggtgg catctttcgg
     3601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3841 ggttacatcg aactggatct caatagtggt aagatccttg agagttttcg ccccgaagaa
     3901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4261 gtaatggtaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     4921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5341 agcaacgcgg cctttttacg gttcctggcc ttttgctgcg gttttgctca catgttcttt
     5401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     5581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     5641 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     5701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag atttaattaa
     5761 gg
//
