LOCUS       sequence-420379-        6965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-tagged Lamin-A specifically in cardiomyocytes by the 
            Tnnt2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6965)
  AUTHORS   Sun Y, Guo C, Chen Z, Lin J, Yang L, Zhang Y, Wu C, Zhao D, Jardin 
            B, Pu WT, Zhao M, Dong E, Hu X, Zhang S, Guo Y
  TITLE     Non-Cell-Autonomous Cardiomyocyte Regulation Complicates Gene 
            Supplementation Therapy for Lmna-Associated Cardiac Defects in Mice.
  JOURNAL   JACC Basic Transl Sci. 2024 Aug 21;9(11):1308-1325. doi: 
            10.1016/j.jacbts.2024.06.004. eCollection 2024 Nov.
  PUBMED    39619139
REFERENCE   2  (bases 1 to 6965)
  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"; doi: 
            "10.1016/j.jacbts.2024.06.004"; journalName: "JACC Basic Transl 
            Sci"; date: "2024-08-21- 21"; volume: "9"; issue: "11"; pages: 
            "1308-1325"
COMMENT     Sequence Label: pAAV-Tnnt2-GFP-LA
FEATURES             Location/Qualifiers
     source          1..6965
                     /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      1022..1031
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1028..1030,1031..1033,1034..1744)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1028..1030,1031..1033,1034..1744)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1073..1094)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1334..1353)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1681..1702
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3802..3821)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3867..3922
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   4015..4144
                     /label=AAV2 ITR
     primer_bind     complement(4163..4180)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4163..4179)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4172..4194)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4321..4776
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4408..4427)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4618..4639
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4802..4906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4907..4975,4976..5767)
                     /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(5125..5144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5938..6526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6427..6446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6814..6835
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6850..6855,6856..6873,6874..6880)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6888..6904
                     /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     6893..6915
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6912..6928
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6912..6928
                     /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 gcctcgagaa gcggccgcac tactccgcgg actactacta gtttcacgcg
     1021 ggccgccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1081 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1141 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1201 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1261 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1321 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1381 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1441 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1501 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1561 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1621 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1681 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1741 caagatggag accccgtccc agcggcgcgc cacccgcagc ggggcgcagg ccagctccac
     1801 tccgctgtcg cccacccgca tcacccggct gcaggagaag gaggacctgc aggagctcaa
     1861 tgatcgcttg gcggtctaca tcgaccgtgt gcgctcgctg gaaacggaga acgcagggct
     1921 gcgccttcgc atcaccgagt ctgaagaggt ggtcagccgc gaggtgtccg gcatcaaggc
     1981 cgcctacgag gccgagctcg gggatgcccg caagaccctt gactcagtag ccaaggagcg
     2041 cgcccgcctg cagctggagc tgagcaaagt gcgtgaggag tttaaggagc tgaaagcgcg
     2101 caataccaag aaggagggtg acctgatagc tgctcaggct cggctgaagg acctggaggc
     2161 tctgctgaac tccaaggagg ccgcactgag cactgctctc agtgagaagc gcacgctgga
     2221 gggcgagctg catgatctgc ggggccaggt ggccaagctt gaggcagccc taggtgaggc
     2281 caagaagcaa cttcaggatg agatgctgcg gcgggtggat gctgagaaca ggctgcagac
     2341 catgaaggag gaactggact tccagaagaa catctacagt gaggagctgc gtgagaccaa
     2401 gcgccgtcat gagacccgac tggtggagat tgacaatggg aagcagcgtg agtttgagag
     2461 ccggctggcg gatgcgctgc aggaactgcg ggcccagcat gaggaccagg tggagcagta
     2521 taagaaggag ctggagaaga cttattctgc caagctggac aatgccaggc agtctgctga
     2581 gaggaacagc aacctggtgg gggctgccca cgaggagctg cagcagtcgc gcatccgcat
     2641 cgacagcctc tctgcccagc tcagccagct ccagaagcag ctggcagcca aggaggcgaa
     2701 gcttcgagac ctggaggact cactggcccg tgagcgggac accagccggc ggctgctggc
     2761 ggaaaaggag cgggagatgg ccgagatgcg ggcaaggatg cagcagcagc tggacgagta
     2821 ccaggagctt ctggacatca agctggccct ggacatggag atccacgcct accgcaagct
     2881 cttggagggc gaggaggaga ggctacgcct gtcccccagc cctacctcgc agcgcagccg
     2941 tggccgtgct tcctctcact catcccagac acagggtggg ggcagcgtca ccaaaaagcg
     3001 caaactggag tccactgaga gccgcagcag cttctcacag cacgcacgca ctagcgggcg
     3061 cgtggccgtg gaggaggtgg atgaggaggg caagtttgtc cggctgcgca acaagtccaa
     3121 tgaggaccag tccatgggca attggcagat caagcgccag aatggagatg atcccttgct
     3181 gacttaccgg ttcccaccaa agttcaccct gaaggctggg caggtggtga cgatctgggc
     3241 tgcaggagct ggggccaccc acagcccccc taccgacctg gtgtggaagg cacagaacac
     3301 ctggggctgc gggaacagcc tgcgtacggc tctcatcaac tccactgggg aagaagtggc
     3361 catgcgcaag ctggtgcgct cagtgactgt ggttgaggac gacgaggatg aggatggaga
     3421 tgacctgctc catcaccacc acggctccca ctgcagcagc tcgggggacc ccgctgagta
     3481 caacctgcgc tcgcgcaccg tgctgtgcgg gacctgcggg cagcctgccg acaaggcatc
     3541 tgccagcggc tcaggagccc aggtgggcgg acccatctcc tctggctctt ctgcctccag
     3601 tgtcacggtc actcgcagct accgcagtgt ggggggcagt gggggtggca gcttcgggga
     3661 caatctggtc acccgctcct acctcctggg caactccagc ccccgaaccc agagccccca
     3721 gaactgcagc atcatgtaag gtacctctag agtcgacccg ggcggcctcg aggacggggt
     3781 gaactacgcc tgaggatccg atctttttcc ctctgccaaa aattatgggg acatcatgaa
     3841 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     3901 ttggaatttt ttgtgtctct cactcggaag caattcgttg atctgaattt cgaccaccca
     3961 taatacccat taccctggta gataagtagc atggcgggtt aatcattaac tacaaggaac
     4021 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4081 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4141 gcagccttaa ttaacctaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4201 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4261 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     4321 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4381 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4441 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4501 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4561 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4621 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4681 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4741 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     4801 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4861 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4921 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4981 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5041 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5101 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     5161 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5221 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5281 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5341 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5401 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     5461 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5521 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5581 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5641 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5701 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5761 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5821 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5881 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5941 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6001 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6061 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6121 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6181 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6241 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6301 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6361 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6421 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6481 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6541 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6601 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6661 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     6721 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     6781 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     6841 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     6901 acaatttcac acaggaaaca gctatgacca tgattacgcc agatttaatt aaggccttaa
     6961 ttagg
//
