LOCUS       sequence-409789-        6793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-tagged human Lamin-A protein by the constitutive CMV 
            promoter in mice.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6793)
  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 6793)
  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-CMV-GFP-LA
FEATURES             Location/Qualifiers
     source          1..6793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        216..595
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        596..799
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      850..859
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(856..858,859..861,862..1572)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(856..858,859..861,862..1572)
                     /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(901..922)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1162..1181)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1509..1530
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3630..3649)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3695..3750
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   3843..3972
                     /label=AAV2 ITR
     primer_bind     complement(3991..4008)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3991..4007)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4000..4022)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4149..4604
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4236..4255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4446..4467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4630..4734
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4735..4803,4804..5595)
                     /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(4953..4972)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5766..6354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6255..6274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6642..6663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6678..6683,6684..6701,6702..6708)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6716..6732
                     /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     6721..6743
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6740..6756
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6740..6756
                     /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 gccagatatc gcgttgacat tgattattga ctagttatta
      241 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      301 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      361 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      421 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      481 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      541 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      601 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      661 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      721 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      781 ggtctatata agcagagctc ctcgagaagc ggccgcacta ctccgcggac tactactagt
      841 ttcacgcggg ccgccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      901 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      961 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1021 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1081 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1141 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1201 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1261 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1321 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1381 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1441 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1501 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1561 gagctgtaca agatggagac cccgtcccag cggcgcgcca cccgcagcgg ggcgcaggcc
     1621 agctccactc cgctgtcgcc cacccgcatc acccggctgc aggagaagga ggacctgcag
     1681 gagctcaatg atcgcttggc ggtctacatc gaccgtgtgc gctcgctgga aacggagaac
     1741 gcagggctgc gccttcgcat caccgagtct gaagaggtgg tcagccgcga ggtgtccggc
     1801 atcaaggccg cctacgaggc cgagctcggg gatgcccgca agacccttga ctcagtagcc
     1861 aaggagcgcg cccgcctgca gctggagctg agcaaagtgc gtgaggagtt taaggagctg
     1921 aaagcgcgca ataccaagaa ggagggtgac ctgatagctg ctcaggctcg gctgaaggac
     1981 ctggaggctc tgctgaactc caaggaggcc gcactgagca ctgctctcag tgagaagcgc
     2041 acgctggagg gcgagctgca tgatctgcgg ggccaggtgg ccaagcttga ggcagcccta
     2101 ggtgaggcca agaagcaact tcaggatgag atgctgcggc gggtggatgc tgagaacagg
     2161 ctgcagacca tgaaggagga actggacttc cagaagaaca tctacagtga ggagctgcgt
     2221 gagaccaagc gccgtcatga gacccgactg gtggagattg acaatgggaa gcagcgtgag
     2281 tttgagagcc ggctggcgga tgcgctgcag gaactgcggg cccagcatga ggaccaggtg
     2341 gagcagtata agaaggagct ggagaagact tattctgcca agctggacaa tgccaggcag
     2401 tctgctgaga ggaacagcaa cctggtgggg gctgcccacg aggagctgca gcagtcgcgc
     2461 atccgcatcg acagcctctc tgcccagctc agccagctcc agaagcagct ggcagccaag
     2521 gaggcgaagc ttcgagacct ggaggactca ctggcccgtg agcgggacac cagccggcgg
     2581 ctgctggcgg aaaaggagcg ggagatggcc gagatgcggg caaggatgca gcagcagctg
     2641 gacgagtacc aggagcttct ggacatcaag ctggccctgg acatggagat ccacgcctac
     2701 cgcaagctct tggagggcga ggaggagagg ctacgcctgt cccccagccc tacctcgcag
     2761 cgcagccgtg gccgtgcttc ctctcactca tcccagacac agggtggggg cagcgtcacc
     2821 aaaaagcgca aactggagtc cactgagagc cgcagcagct tctcacagca cgcacgcact
     2881 agcgggcgcg tggccgtgga ggaggtggat gaggagggca agtttgtccg gctgcgcaac
     2941 aagtccaatg aggaccagtc catgggcaat tggcagatca agcgccagaa tggagatgat
     3001 cccttgctga cttaccggtt cccaccaaag ttcaccctga aggctgggca ggtggtgacg
     3061 atctgggctg caggagctgg ggccacccac agccccccta ccgacctggt gtggaaggca
     3121 cagaacacct ggggctgcgg gaacagcctg cgtacggctc tcatcaactc cactggggaa
     3181 gaagtggcca tgcgcaagct ggtgcgctca gtgactgtgg ttgaggacga cgaggatgag
     3241 gatggagatg acctgctcca tcaccaccac ggctcccact gcagcagctc gggggacccc
     3301 gctgagtaca acctgcgctc gcgcaccgtg ctgtgcggga cctgcgggca gcctgccgac
     3361 aaggcatctg ccagcggctc aggagcccag gtgggcggac ccatctcctc tggctcttct
     3421 gcctccagtg tcacggtcac tcgcagctac cgcagtgtgg ggggcagtgg gggtggcagc
     3481 ttcggggaca atctggtcac ccgctcctac ctcctgggca actccagccc ccgaacccag
     3541 agcccccaga actgcagcat catgtaaggt acctctagag tcgacccggg cggcctcgag
     3601 gacggggtga actacgcctg aggatccgat ctttttccct ctgccaaaaa ttatggggac
     3661 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     3721 atagtgtgtt ggaatttttt gtgtctctca ctcggaagca attcgttgat ctgaatttcg
     3781 accacccata atacccatta ccctggtaga taagtagcat ggcgggttaa tcattaacta
     3841 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3901 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3961 gcgagcgcgc agccttaatt aacctaattc actggccgtc gttttacaac gtcgtgactg
     4021 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4081 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4141 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4201 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4261 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4321 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4381 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4441 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4501 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4561 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     4621 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4681 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4741 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4801 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4861 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4921 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4981 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5041 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5101 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5161 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5221 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5281 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5341 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5401 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5461 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5521 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5581 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5641 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5701 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5761 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5821 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5881 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5941 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6001 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6061 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6121 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6181 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6241 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6301 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6361 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6421 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6481 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6541 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6601 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6661 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6721 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccag atttaattaa
     6781 ggccttaatt agg
//
