LOCUS       sequence-326315-        7257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-based delivery of dnMRTFA-GFP into cardiomyocytes in vivo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7257)
  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 7257)
  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-dnmrtfa-gfp
FEATURES             Location/Qualifiers
     source          1..7257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          772..904
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     887..906
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        949..967
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3295..3304
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3301..3303,3304..3306,3307..4020)
                     /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(3301..3303,3304..3306,3307..4017)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3346..3367)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3607..3626)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3954..3975
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4094..4113)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4159..4214
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4307..4436
                     /label=AAV2 ITR
     primer_bind     complement(4455..4472)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4455..4471)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4464..4486)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4613..5068
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4700..4719)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4910..4931
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5094..5198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5199..5267,5268..6059)
                     /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(5417..5436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6230..6818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6719..6738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7106..7127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7142..7147,7148..7165,7166..7172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7180..7196
                     /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     7185..7207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7204..7220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7204..7220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg
      181 atcctctaga actatagcta gaattcgccc ttacgggccc cccctcgagg tcgggataaa
      241 agcagtctgg gctttcacat gacagcatct ggggctgcgg cagagggtcg ggtccgaagc
      301 gctgccttat cagcgtcccc agccctggga ggtgacagct ggctggcttg tgtcagcccc
      361 tcgggcactc acgtatctcc gtccgacggg tttaaaatag caaaactctg aggccacaca
      421 atagcttggg cttatatggg ctcctgtggg ggaaggggga gcacggaggg ggccggggcc
      481 gctgctgcca aaatagcagc tcacaagtgt tgcattcctc tctgggcgcc gggcacattc
      541 ctgctggctc tgcccgcccc ggggtgggcg ccggggggac cttaaagcct ctgcccccca
      601 aggagccctt cccagacagc cgccggcacc caccgctccg tgggacgatc cccgaagctc
      661 tagagcttta ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac
      721 acaacagtct cgaacttaag ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc
      781 aaggttacaa gacaggttta aggagaccaa tagaaactgg gcttgtcgag acagagaaga
      841 ctcttgcgtt tctgataggc acctattggt cttactgaca tccactttgc ctttctctcc
      901 acaggtgtcc actcccagtt caattacagc tcttaaggct agagtactta atacgactca
      961 ctataggcta gcatgactct gctggagcct gagatgctaa tgatggctgt gcagtcagtg
     1021 ctgcagttga agctccagca gcggcggacc cgggaggaac tggtgagcca agggatcatg
     1081 ccgcctttga aaagccccgc tgcatttcat gagcagagaa gaagcctgga gcgggccagg
     1141 accgaggact atttgaaacg gaagatccgt tcccggcccg agagagcaga gctggtcagg
     1201 atgcacattc tggaagagac ctcggctgag ccttcgctcc aggccaagca gctgaagctg
     1261 aagagagcca ggctggctga tgacctcaat gaaaagattg cacagaggcc tggccccatg
     1321 gagctggtgg agaagaatat cctgcctgtg gagtccagcc tgaaggaggc tatcattgtg
     1381 ggccaggtaa attacccaaa ggtagcagac agttcctcct tcgacgagga cagcagcgat
     1441 gccctgtctc ctgagcagcc tgccagccat gagtcccagg gttcagtgcc atcacccttg
     1501 gagtcccgag tcagtgatcc actgcccagt gccacctcca tatcacccac tcaggttctt
     1561 tctcaactcc caatggctcc ggatcctgga gagacgcttt ttctggcaga gcagcctcct
     1621 ctgcctcccg cacctctgct gcccccaagc ctagccaatg gaagcatcgt ccccactgcc
     1681 aagcctgctc ccacactcat caagcaaagc caacccaagt ctgccagcga gaaatcacag
     1741 cgcagcaaga aggccaagga gctgaagcca aaggtgaaga agctcaagta ccaccagtac
     1801 atccccccgg accagaagca ggacaagggg gcgcccgcca tggactcctc ctatgccaag
     1861 atcctgcagc agcagcagct cttcctgcag ctgcagatcc tcaaccagca gcagcagcag
     1921 cagcagcaac agcactacaa ctaccaggcc atcctgcctg cccctcccaa gccctcggct
     1981 gagactcctg gaagcagtgc ccctacccca tcacgcagcc tctccaccag tagcagcccc
     2041 agctcaggca ccccagggcc cagcgggctg gcacgccaga gcagcaccgc actagctgcc
     2101 aaaccaggag ccctgccagc caacctggat gacatgaagg tggcagagct gaagcaagaa
     2161 ctgaagttgc ggtcccttcc cgtctcaggc accaagacag agctgataga acgcctgcgt
     2221 gcctaccaag accaagtcag cccagctcca ggagccccca aggcccctgc caccacctct
     2281 gtgctgtcca aggctggtga ggtagtggtc gccttccctg cggccctgct aagcacaggg
     2341 tcagctcttg taacagcagg ccttgcacca gctgagatgg tggtggccac agtaaccagc
     2401 aatggcatgg tgaagtttgg cagcacaggc tccacacccc ccgtgtctcc caccccttca
     2461 gagcgctcac tgctcagcac gggtgatgag aattccacac ctggggatgc ctttggtgaa
     2521 atggtgacat cgccgctgac acagctcacc ctgcaggcct ccccactgca gatcgtgaag
     2581 gaggagggtg cccgtgctgc gtcctgctgt ctaagccctg gtgctcgggc tgagctggag
     2641 ggactggaca aggaccagat gctgcaggag aaggacaagc agattgagga gctgacccga
     2701 atgctccaac agaagcagca gctggttgag ctgctgcggc tacagctgga gcagcagaag
     2761 cgggcccagc agccagcccc agccagcagc cctgtgaaga gggaaagtgg tttctccagt
     2821 tgccagctga gctgccagcc ccagggctct gcccatgctt ttggctctgg cctagtggtt
     2881 cccactacca accatggaga cactcaggcc ccagcgccag agtccccacc tgtggtggtg
     2941 aagcaggaag ctgggccacc tgagccagat ctggccccca gctcccagct gctcttgggc
     3001 tcccagggca ccagcttcct caagagggtc agccctccta ccctggtcac tgactctaca
     3061 gggactcacc tcatcctcac tgtgaccaat aagagtgctg atggccctgg cttgcctgca
     3121 gggagccccc agcagccctt gtcccagcct ggttctccag cccctggtcc acctgcccag
     3181 atggacctgg agcacccacc tcagcctccg tttgcaaccc ccacatctct gctgaagaag
     3241 gagccccctg gttatgaaga gactgtgccg cggactacta ctagtttcac gcgggccgcc
     3301 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3361 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3421 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3481 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3541 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3601 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3661 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3721 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3781 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3841 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3901 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3961 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     4021 taagctcgcg tggtacctct agagtcgacc cgggcggcct cgaggacggg gtgaactacg
     4081 cctgaggatc cgatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     4141 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     4201 ttttgtgtct ctcactcgga agcaattcgt tgatctgaat ttcgaccacc cataataccc
     4261 attaccctgg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     4321 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4381 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt
     4441 aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     4501 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     4561 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     4621 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4681 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4741 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4801 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     4861 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     4921 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4981 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5041 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     5101 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5161 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5221 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5281 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5341 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5401 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5461 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5521 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5581 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5641 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5701 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5761 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5821 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5881 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5941 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6001 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6061 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6121 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6181 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6241 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6301 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6361 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6421 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6481 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6541 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6601 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6661 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6721 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6781 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6841 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6901 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6961 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     7021 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     7081 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     7141 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     7201 acacaggaaa cagctatgac catgattacg ccagatttaa ttaaggcctt aattagg
//
