LOCUS       sequence-405801-        4369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4369)
  AUTHORS   Gray JT, Zolotukhin S
  TITLE     Design and Construction of Functional AAV Vectors.
  JOURNAL   Methods Mol Biol. 2011;807:25-46.
  PUBMED    22034025
REFERENCE   2  (bases 1 to 4369)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2011"; volume: "807"; pages: "25-46"
COMMENT     Sequence Label: pscAAV-GFP
FEATURES             Location/Qualifiers
     source          1..4369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   81..210
                     /label=AAV2 ITR
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     567..591
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      732..741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(738..740,741..743,744..1457)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(738..740,741..743,744..1454)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(783..804)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1044..1063)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1391..1412
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1501..1520
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1770..1787)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1770..1786)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1779..1801)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1995..2014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2114..2136
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2174..2192)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2260..2364
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2365..2433,2434..3225)
                     /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"
     primer_bind     complement(2583..2602)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3396..3984
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3885..3904
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4272..4293
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4308..4313,4314..4331,4332..4338)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4346..4362
                     /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     join(4351..4369,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agctctcgag atctagaaag cttcccgggg
       61 ggatctgggc cactccctct ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
      121 ccgggcgtcg ggcgaccaaa ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      181 gagtggccaa ctccatcact aggggttcct ggaggggtgg agtcgtgacc taggcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
      601 cgccatccgg actctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg
      661 attctaattg tttctctctt ttagattcca acctttggaa ctgaattccg cgggcccggg
      721 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
      781 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
      841 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
      901 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
      961 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     1021 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     1081 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1141 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     1201 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     1261 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     1321 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     1381 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     1441 acgagctgta caagtaaagc ggccgctagg cctcacctgc gatctcgatg ctttatttgt
     1501 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1561 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1621 actagtccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     1681 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agagagggac
     1741 agatccgggc ccgcatgcgt cgacaattca ctggccgtcg ttttacaacg tcgtgactgg
     1801 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1861 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1921 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     1981 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     2041 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     2101 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     2161 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     2221 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2281 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2341 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2401 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2461 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2521 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2581 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2641 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2701 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2761 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2821 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2881 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2941 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3001 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3061 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3121 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3181 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3241 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3301 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3361 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3421 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3481 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3541 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3601 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3661 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3721 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3781 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3841 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3901 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3961 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4021 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4081 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4141 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     4201 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     4261 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     4321 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcaca
//
