LOCUS       sequence-154392-        6891 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 6891)
  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 6891)
  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: pAAV-GFP
FEATURES             Location/Qualifiers
     source          1..6891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   168..297
                     /label=AAV2 ITR
     primer_bind     complement(353..372)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        474..853
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        854..1057
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1008..1028
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1054..1078
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1171..1646
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     regulatory      1724..1733
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1730..1732,1733..1735,1736..2449)
                     /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(1730..1732,1733..1735,1736..2446)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1775..1796)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2036..2055)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2383..2404
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2479..2495)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2480..2496)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2589..2983
                     /label=beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     repeat_region   3338..3467
                     /label=AAV2 ITR
     rep_origin      3838..4351
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3925..3944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4135..4156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4833..4852)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4952..4974
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5012..5030)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5098..5202
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5203..5271,5272..6063)
                     /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(5421..5440)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6234..6822
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6723..6742
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
       61 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      121 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg cgcgctcgct
      181 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
      241 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt
      301 agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagc ggcctcggcc
      361 tctgcataaa taaaaaaaat tagtcagcca tgagcttggc ccattgcata cgttgtatcc
      421 atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg
      481 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      541 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      601 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      661 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      721 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      781 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      841 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      901 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      961 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     1021 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     1081 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
     1141 cccctcgaag ctgatcctga gaacttcagg gtgagtctat gggacccttg atgttttctt
     1201 tccccttctt ttctatggtt aagttcatgt cataggaagg ggagaagtaa cagggtacac
     1261 atattgacca aatcagggta attttgcatt tgtaatttta aaaaatgctt tcttctttta
     1321 atatactttt ttgtttatct tatttctaat actttcccta atctctttct ttcagggcaa
     1381 taatgataca atgtatcatg cctctttgca ccattctaaa gaataacagt gataatttct
     1441 gggttaaggc aatagcaata tttctgcata taaatatttc tgcatataaa ttgtaactga
     1501 tgtaagaggt ttcatattgc taatagcagc tacaatccag ctaccattct gcttttattt
     1561 tatggttggg ataaggctgg attattctga gtccaagcta ggcccttttg ctaatcatgt
     1621 tcatacctct tatcttcctc ccacagctcc tgggcaacgt gctggtctgt gtgctggccc
     1681 atcactttgg caaagaattc cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa
     1741 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1801 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1861 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1921 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1981 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     2041 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2101 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2161 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2221 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2281 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2341 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2401 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgctc
     2461 tagaggatcc aagcttatcg ataccgtcga cctcgagggc ccagatctaa ttcaccccac
     2521 cagtgcaggc tgcctatcag aaagtggtgg ctggtgtggc taatgccctg gcccacaagt
     2581 atcactaagc tcgctttctt gctgtccaat ttctattaaa ggttcctttg ttccctaagt
     2641 ccaactacta aactggggga tattatgaag ggccttgagc atctggattc tgcctaataa
     2701 aaaacattta ttttcattgc aatgatgtat ttaaattatt tctgaatatt ttactaaaaa
     2761 gggaatgtgg gaggtcagtg catttaaaac ataaagaaat gaagagctag ttcaaacctt
     2821 gggaaaatac actatatctt aaactccatg aaagaaggtg aggctgcaaa cagctaatgc
     2881 acattggcaa cagcccctga tgcctatgcc ttattcatcc ctcagaaaag gattcaagta
     2941 gaggcttgat ttggaggtta aagttttgct atgctgtatt ttacattact tattgtttta
     3001 gctgtcctca tgaatgtctt ttcactaccc atttgcttat cctgcatctc tcagccttga
     3061 ctccactcag ttctcttgct tagagatacc acctttcccc tgaagtgttc cttccatgtt
     3121 ttacggcgag atggtttctc ctcgcctggc cactcagcct tagttgtctc tgttgtctta
     3181 tagaggtcta cttgaagaag gaaaaacagg gggcatggtt tgactgtcct gtgagccctt
     3241 cttccctgcc tcccccactc acagtgaccc ggaatccctc gacatctaga gcatggctac
     3301 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     3361 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3421 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg gcgtaatagc
     3481 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcga
     3541 ttccgttgca atggctggcg gtaatattgt tctggatatt accagcaagg ccgatagttt
     3601 gagttcttct actcaggcaa gtgatgttat tactaatcaa agaagtattg cgacaacggt
     3661 taatttgcgt gatggacaga ctcttttact cggtggcctc actgattata aaaacacttc
     3721 tcaggattct ggcgtaccgt tcctgtctaa aatcccttta atcggcctcc tgtttagctc
     3781 ccgctctgat tctaacgagg aaagcacgtt atacgtgctc gtcaaagcaa ccatagtacg
     3841 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3901 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3961 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4021 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4081 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4141 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4201 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4261 cgaattttaa caaaatatta acgcttacaa tttaaatatt tgcttataca atcttcctgt
     4321 ttttggggct tttctgatta tcaaccgggg tacatatgat tgacatgcta gttttacgat
     4381 taccgttcat cgattctctt gtttgctcca gactctcagg caatgacctg atagcctttg
     4441 tagagacctc tcaaaaatag ctaccctctc cggcatgaat ttatcagcta gaacggttga
     4501 atatcatatt gatggtgatt tgactgtctc cggcctttct cacccgtttg aatctttacc
     4561 tacacattac tcaggcattg catttaaaat atatgagggt tctaaaaatt tttatccttg
     4621 cgttgaaata aaggcttctc ccgcaaaagt attacagggt cataatgttt ttggtacaac
     4681 cgatttagct ttatgctctg aggctttatt gcttaatttt gctaattctt tgccttgcct
     4741 gtatgattta ttggatgttg gaatcgcctg atgcggtatt ttctccttac gcatctgtgc
     4801 ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4861 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4921 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4981 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5041 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5101 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5161 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5221 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5281 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5341 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5401 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5461 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5521 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5581 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5641 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5701 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5761 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5821 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5881 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5941 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6001 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6061 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6121 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6181 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6241 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6301 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6361 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6421 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6481 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6541 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6601 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6661 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6721 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6781 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6841 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc t
//
