LOCUS       sequence-326326-        7832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-based delivery of wtMRTFA-GFP into cardiomyocytes in vivo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7832)
  AUTHORS   Guo Y, Cao Y, Jardin BD, Sethi I, Ma Q, Moghadaszadeh B, Troiano EC,
            Mazumdar N, Trembley MA, Small EM, Yuan GC, Beggs AH, Pu WT
  TITLE     Sarcomeres regulate murine cardiomyocyte maturation through MRTF-SRF
            signaling.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Jan 12;118(2). pii: 2008861118. doi: 
            10.1073/pnas.2008861118.
  PUBMED    33361330
REFERENCE   2  (bases 1 to 7832)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-01-12"; volume: "118(2). pii: 
            2008861118. doi"; pages: " 10.1073/pnas.2008861118"
COMMENT     Sequence Label: aav-tnt-wtmrtfa-gfp
FEATURES             Location/Qualifiers
     source          1..7832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          750..882
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     865..884
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        927..945
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3870..3879
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3876..3878,3879..3881,3882..4595)
                     /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(3876..3878,3879..3881,3882..4592)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3921..3942)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4182..4201)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4529..4550
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4669..4688)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4734..4789
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4882..5011
                     /label=AAV2 ITR
     primer_bind     complement(5030..5047)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5030..5046)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5039..5061)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5188..5643
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5275..5294)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5485..5506
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5669..5773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5774..5842,5843..6634)
                     /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(5992..6011)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6805..7393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7294..7313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7681..7702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7717..7722,7723..7740,7741..7747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7755..7771
                     /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     7760..7782
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7779..7795
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7779..7795
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcggcc tcagtgagcg
       61 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga
      121 ttaacccgcc atgctactta tctaccaggg taatggggat cctctagaac tatagctaga
      181 attcgccctt acgggccccc cctcgaggtc gggataaaag cagtctgggc tttcacatga
      241 cagcatctgg ggctgcggca gagggtcggg tccgaagcgc tgccttatca gcgtccccag
      301 ccctgggagg tgacagctgg ctggcttgtg tcagcccctc gggcactcac gtatctccgt
      361 ccgacgggtt taaaatagca aaactctgag gccacacaat agcttgggct tatatgggct
      421 cctgtggggg aagggggagc acggaggggg ccggggccgc tgctgccaaa atagcagctc
      481 acaagtgttg cattcctctc tgggcgccgg gcacattcct gctggctctg cccgccccgg
      541 ggtgggcgcc ggggggacct taaagcctct gccccccaag gagcccttcc cagacagccg
      601 ccggcaccca ccgctccgtg ggacgatccc cgaagctcta gagctttatt gcggtagttt
      661 atcacagtta aattgctaac gcagtcagtg cttctgacac aacagtctcg aacttaagct
      721 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
      781 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
      841 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca
      901 attacagctc ttaaggctag agtacttaat acgactcact ataggctagc atgactctgc
      961 tggagcctga gatgctaatg atggctgtgc agtcagtgct gcagttgaag ctccagcagc
     1021 ggcggacccg ggaggaactg gtgagccaag ggatcatgcc gcctttgaaa agccccgctg
     1081 catttcatga gcagagaaga agcctggagc gggccaggac cgaggactat ttgaaacgga
     1141 agatccgttc ccggcccgag agagcagagc tggtcaggat gcacattctg gaagagacct
     1201 cggctgagcc ttcgctccag gccaagcagc tgaagctgaa gagagccagg ctggctgatg
     1261 acctcaatga aaagattgca cagaggcctg gccccatgga gctggtggag aagaatatcc
     1321 tgcctgtgga gtccagcctg aaggaggcta tcattgtggg ccaggtaaat tacccaaagg
     1381 tagcagacag ttcctccttc gacgaggaca gcagcgatgc cctgtctcct gagcagcctg
     1441 ccagccatga gtcccagggt tcagtgccat cacccttgga gtcccgagtc agtgatccac
     1501 tgcccagtgc cacctccata tcacccactc aggttctttc tcaactccca atggctccgg
     1561 atcctggaga gacgcttttt ctggcagagc agcctcctct gcctcccgca cctctgctgc
     1621 ccccaagcct agccaatgga agcatcgtcc ccactgccaa gcctgctccc acactcatca
     1681 agcaaagcca acccaagtct gccagcgaga aatcacagcg cagcaagaag gccaaggagc
     1741 tgaagccaaa ggtgaagaag ctcaagtacc accagtacat ccccccggac cagaagcagg
     1801 acaagggggc gcccgccatg gactcctcct atgccaagat cctgcagcag cagcagctct
     1861 tcctgcagct gcagatcctc aaccagcagc agcagcagca gcagcaacag cactacaact
     1921 accaggccat cctgcctgcc cctcccaagc cctcggctga gactcctgga agcagtgccc
     1981 ctaccccatc acgcagcctc tccaccagta gcagccccag ctcaggcacc ccagggccca
     2041 gcgggctggc acgccagagc agcaccgcac tagctgccaa accaggagcc ctgccagcca
     2101 acctggatga catgaaggtg gcagagctga agcaagaact gaagttgcgg tcccttcccg
     2161 tctcaggcac caagacagag ctgatagaac gcctgcgtgc ctaccaagac caagtcagcc
     2221 cagctccagg agcccccaag gcccctgcca ccacctctgt gctgtccaag gctggtgagg
     2281 tagtggtcgc cttccctgcg gccctgctaa gcacagggtc agctcttgta acagcaggcc
     2341 ttgcaccagc tgagatggtg gtggccacag taaccagcaa tggcatggtg aagtttggca
     2401 gcacaggctc cacacccccc gtgtctccca ccccttcaga gcgctcactg ctcagcacgg
     2461 gtgatgagaa ttccacacct ggggatgcct ttggtgaaat ggtgacatcg ccgctgacac
     2521 agctcaccct gcaggcctcc ccactgcaga tcgtgaagga ggagggtgcc cgtgctgcgt
     2581 cctgctgtct aagccctggt gctcgggctg agctggaggg actggacaag gaccagatgc
     2641 tgcaggagaa ggacaagcag attgaggagc tgacccgaat gctccaacag aagcagcagc
     2701 tggttgagct gctgcggcta cagctggagc agcagaagcg ggcccagcag ccagccccag
     2761 ccagcagccc tgtgaagagg gaaagtggtt tctccagttg ccagctgagc tgccagcccc
     2821 agggctctgc ccatgctttt ggctctggcc tagtggttcc cactaccaac catggagaca
     2881 ctcaggcccc agcgccagag tccccacctg tggtggtgaa gcaggaagct gggccacctg
     2941 agccagatct ggcccccagc tcccagctgc tcttgggctc ccagggcacc agcttcctca
     3001 agagggtcag ccctcctacc ctggtcactg actctacagg gactcacctc atcctcactg
     3061 tgaccaataa gagtgctgat ggccctggct tgcctgcagg gagcccccag cagcccttgt
     3121 cccagcctgg ttctccagcc cctggtccac ctgcccagat ggacctggag cacccacctc
     3181 agcctccgtt tgcaaccccc acatctctgc tgaagaagga gccccctggt tatgaagaga
     3241 ctgtgaccca gcagcctaag cagcaggaaa atggctcctc cagtcagcac atggatgatc
     3301 tgtttgatat tcttattcag agtggagaga tttcagcaga tttcaaagag ccaccatccc
     3361 taccaggcaa ggaaaagtca cctccagcag cagcagcgta tgggcctcca ttgacaccac
     3421 aaccctcgcc tttgagtgaa ctcccccaag ctgctcctcc accaggttcc cccaccctcc
     3481 cagggcgcct tgaagacttc ctggagagca gcacagggct gcccctgctg acaagtgggc
     3541 acgagggacc agaacccctt tccctcattg atgacctcca cagccagatg ctgagcagct
     3601 ccgccatcct ggaccacccc ccatcaccca tggacacctc tgaattgcac tttgctcctg
     3661 agcccagcag tggtatgggc ctggacctgg ctgttggcca cctggacagc atggactggc
     3721 tggagctgtc gtctggtggc cctgtgctca gcctggctcc cctcagcact gcagccccca
     3781 gcctcttctc gatggacttc ctggatggac acgacttgca gctccactgg gattcctgct
     3841 tgccgcggac tactactagt ttcacgcggg ccgccatggt gagcaagggc gaggagctgt
     3901 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     3961 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     4021 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     4081 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     4141 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     4201 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     4261 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     4321 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     4381 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     4441 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     4501 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     4561 ggatcactct cggcatggac gagctgtaca agtaataagc tcgcgtggta cctctagagt
     4621 cgacccgggc ggcctcgagg acggggtgaa ctacgcctga ggatccgatc tttttccctc
     4681 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     4741 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaagcaa
     4801 ttcgttgatc tgaatttcga ccacccataa tacccattac cctggtagat aagtagcatg
     4861 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     4921 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4981 gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg
     5041 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     5101 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5161 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     5221 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5281 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5341 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5401 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5461 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5521 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5581 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5641 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5701 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5761 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5821 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5881 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5941 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6001 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6061 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6121 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6181 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6241 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6301 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6361 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6421 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6481 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6541 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6601 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6661 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6721 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6781 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6841 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6901 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6961 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7021 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7081 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7141 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7201 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7261 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7321 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7381 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7441 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7501 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7561 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     7621 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     7681 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     7741 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     7801 ttacgccaga tttaattaag gccttaatta gg
//
