LOCUS       sequence-409795-        6847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-lamin-A only in non-myocyte when delivered into mice 
            carrying a cardiomyocyte-specific Myh6-Cre transgene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6847)
  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 6847)
  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-Loxp-CMV-Loxp-GFP-LA
FEATURES             Location/Qualifiers
     source          1..6847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    198..231
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        251..630
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        631..834
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    835..868
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      904..913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(910..912,913..915,916..1626)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(910..912,913..915,916..1626)
                     /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(955..976)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1216..1235)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1563..1584
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3684..3703)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3749..3804
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   3897..4026
                     /label=AAV2 ITR
     primer_bind     complement(4045..4062)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4045..4061)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4054..4076)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4203..4658
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4290..4309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4500..4521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4684..4788
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4789..4857,4858..5649)
                     /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(5007..5026)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5820..6408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6309..6328
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6696..6717
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6732..6737,6738..6755,6756..6762)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6770..6786
                     /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     6775..6797
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6794..6810
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6794..6810
                     /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 actatagata acttcgtata atgtatgcta tacgaagtta tctagccaga
      241 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
      301 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      361 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      421 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      481 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      541 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      601 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      661 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      721 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      781 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctataact
      841 tcgtataatg tatgctatac gaagttatgc actactccgc ggactactac tagtttcacg
      901 cgggccgcca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      961 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     1021 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     1081 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     1141 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1201 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1261 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1321 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1381 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1441 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1501 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     1561 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1621 tacaagatgg agaccccgtc ccagcggcgc gccacccgca gcggggcgca ggccagctcc
     1681 actccgctgt cgcccacccg catcacccgg ctgcaggaga aggaggacct gcaggagctc
     1741 aatgatcgct tggcggtcta catcgaccgt gtgcgctcgc tggaaacgga gaacgcaggg
     1801 ctgcgccttc gcatcaccga gtctgaagag gtggtcagcc gcgaggtgtc cggcatcaag
     1861 gccgcctacg aggccgagct cggggatgcc cgcaagaccc ttgactcagt agccaaggag
     1921 cgcgcccgcc tgcagctgga gctgagcaaa gtgcgtgagg agtttaagga gctgaaagcg
     1981 cgcaatacca agaaggaggg tgacctgata gctgctcagg ctcggctgaa ggacctggag
     2041 gctctgctga actccaagga ggccgcactg agcactgctc tcagtgagaa gcgcacgctg
     2101 gagggcgagc tgcatgatct gcggggccag gtggccaagc ttgaggcagc cctaggtgag
     2161 gccaagaagc aacttcagga tgagatgctg cggcgggtgg atgctgagaa caggctgcag
     2221 accatgaagg aggaactgga cttccagaag aacatctaca gtgaggagct gcgtgagacc
     2281 aagcgccgtc atgagacccg actggtggag attgacaatg ggaagcagcg tgagtttgag
     2341 agccggctgg cggatgcgct gcaggaactg cgggcccagc atgaggacca ggtggagcag
     2401 tataagaagg agctggagaa gacttattct gccaagctgg acaatgccag gcagtctgct
     2461 gagaggaaca gcaacctggt gggggctgcc cacgaggagc tgcagcagtc gcgcatccgc
     2521 atcgacagcc tctctgccca gctcagccag ctccagaagc agctggcagc caaggaggcg
     2581 aagcttcgag acctggagga ctcactggcc cgtgagcggg acaccagccg gcggctgctg
     2641 gcggaaaagg agcgggagat ggccgagatg cgggcaagga tgcagcagca gctggacgag
     2701 taccaggagc ttctggacat caagctggcc ctggacatgg agatccacgc ctaccgcaag
     2761 ctcttggagg gcgaggagga gaggctacgc ctgtccccca gccctacctc gcagcgcagc
     2821 cgtggccgtg cttcctctca ctcatcccag acacagggtg ggggcagcgt caccaaaaag
     2881 cgcaaactgg agtccactga gagccgcagc agcttctcac agcacgcacg cactagcggg
     2941 cgcgtggccg tggaggaggt ggatgaggag ggcaagtttg tccggctgcg caacaagtcc
     3001 aatgaggacc agtccatggg caattggcag atcaagcgcc agaatggaga tgatcccttg
     3061 ctgacttacc ggttcccacc aaagttcacc ctgaaggctg ggcaggtggt gacgatctgg
     3121 gctgcaggag ctggggccac ccacagcccc cctaccgacc tggtgtggaa ggcacagaac
     3181 acctggggct gcgggaacag cctgcgtacg gctctcatca actccactgg ggaagaagtg
     3241 gccatgcgca agctggtgcg ctcagtgact gtggttgagg acgacgagga tgaggatgga
     3301 gatgacctgc tccatcacca ccacggctcc cactgcagca gctcggggga ccccgctgag
     3361 tacaacctgc gctcgcgcac cgtgctgtgc gggacctgcg ggcagcctgc cgacaaggca
     3421 tctgccagcg gctcaggagc ccaggtgggc ggacccatct cctctggctc ttctgcctcc
     3481 agtgtcacgg tcactcgcag ctaccgcagt gtggggggca gtgggggtgg cagcttcggg
     3541 gacaatctgg tcacccgctc ctacctcctg ggcaactcca gcccccgaac ccagagcccc
     3601 cagaactgca gcatcatgta aggtacctct agagtcgacc cgggcggcct cgaggacggg
     3661 gtgaactacg cctgaggatc cgatcttttt ccctctgcca aaaattatgg ggacatcatg
     3721 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     3781 tgttggaatt ttttgtgtct ctcactcgga agcaattcgt tgatctgaat ttcgaccacc
     3841 cataataccc attaccctgg tagataagta gcatggcggg ttaatcatta actacaagga
     3901 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3961 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4021 gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4081 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4141 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4201 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4261 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4321 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4381 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4441 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4501 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4561 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4621 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4681 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4741 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4801 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4861 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4921 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4981 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5041 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5101 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5161 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5221 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5281 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5341 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5401 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5461 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5521 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5581 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5641 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5701 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5761 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5821 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5881 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5941 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6001 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6061 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6121 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6181 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6241 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6301 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6361 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6421 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6481 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6541 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6601 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     6661 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     6721 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     6781 taacaatttc acacaggaaa cagctatgac catgattacg ccagatttaa ttaaggcctt
     6841 aattagg
//
