LOCUS       sequence-275779-        5010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses GFP under the promoter of CAG. Transgene flanked by AAV2 
            ITR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5010)
  AUTHORS   Wang S, Li Y, Xu Y, Ma Q, Lin Z, Schlame M, Bezzerides VJ, Strathdee
            D, Pu WT
  TITLE     AAV Gene Therapy Prevents and Reverses Heart Failure in A Murine 
            Knockout Model of Barth Syndrome.
  JOURNAL   Circ Res. 2020 Mar 9. doi: 10.1161/CIRCRESAHA.119.315956.
  PUBMED    32146862
REFERENCE   2  (bases 1 to 5010)
  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: "Circ Res. 
            2020 Mar 9. doi: 10.1161/CIRCRESAHA.119.315956."
COMMENT     Sequence Label: pCAG/GFP
FEATURES             Location/Qualifiers
     source          1..5010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        134..411
                     /label=chicken beta-actin promoter
     primer_bind     690..712
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     774..793
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(889..891,892..894,895..1608)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSTMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(934..955)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1195..1214)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1542..1563
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1682..1701)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1778..1899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1789..1808)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1843..1862
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2061..2190
                     /label=AAV2 ITR
     rep_origin      complement(2427..3015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2507..2526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3186..3977,3978..4046))
                     /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     3809..3828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4047..4151)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4177..4632)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4314..4335)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4526..4545
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4704..4833
                     /label=AAV2 ITR
ORIGIN
        1 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
       61 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      121 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
      181 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
      241 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
      301 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
      361 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
      421 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
      481 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
      541 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
      601 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggctgtccgc ggggggacgg
      661 ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc
      721 tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt
      781 gctggttatt gtgctgtctc atcattttgg caaagaattg gatccccggg taccgctagc
      841 tctctgttaa agcaagcagg agacgtggaa gaaaaccccg gtcctgccat ggtgagcaag
      901 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      961 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1021 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1081 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1141 ttcaagtcca ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1201 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1261 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1321 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1381 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1441 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1501 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1561 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaata agctcgcgtg
     1621 gtacctctag agtcgacccg ggcggcctcg aggacggggt gaactacgcc tgaggatccg
     1681 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     1741 ctggctaata aagggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     1801 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     1861 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     1921 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     1981 aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg
     2041 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     2101 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     2161 ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg
     2221 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2281 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2341 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2401 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2461 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2521 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2581 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2641 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2701 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2761 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2821 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2881 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2941 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3001 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3061 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3121 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3181 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3241 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3301 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3361 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3421 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3481 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3541 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3601 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3661 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3721 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3781 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3841 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3901 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3961 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4021 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4081 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4141 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     4201 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4261 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4321 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4381 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4441 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4501 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4561 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4621 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4681 gggcctcttc gctattacgc cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     4741 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     4801 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
     4861 acttatctac gtagccatgc tctaggaaga gtaccattga cgtcaataat gacgtatgtt
     4921 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     4981 actgcccact tggcagtaca tcaagtgtat
//
