LOCUS       sequence-429370-        6652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  cardiomyocytes-specific plasmid, expressing Gfp and the coding 
            sequence of mouse Palmd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6652)
  AUTHORS   Guo CT, Jardin BD, Lin JS, Ambroise RL, Wang Z, Yang LZ, Mazumdar N,
            Lu FJ, Ma Q, Cao YP, Liu CZ, Li KL, Liu XJ, Lan F, Zhao MM, Xiao H, 
            Dong ED, Pu WT, Guo YX
  TITLE     In vivo proximity proteomics uncovers palmdelphin (PALMD) as a 
            Z-disc-associated mitigator of isoproterenol-induced cardiac injury.
  JOURNAL   Acta Pharmacol Sin. 2024 Jul 23. doi: 10.1038/s41401-024-01348-y.
  PUBMED    39043970
REFERENCE   2  (bases 1 to 6652)
  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: "Acta 
            Pharmacol Sin. 2024 Jul 23. doi: 10.1038/s41401-024-01348-y."
COMMENT     Sequence Label: pAAV-cTNT-GFP-Palmd
FEATURES             Location/Qualifiers
     source          1..6652
                     /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"
     CDS             join(1013..1015,1016..1018,1019..1729)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1013..1015,1016..1018,1019..1729)
                     /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(1058..1079)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1319..1338)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1666..1687
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3489..3508)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3554..3609
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   3702..3831
                     /label=AAV2 ITR
     primer_bind     complement(3850..3867)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3850..3866)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3859..3881)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4008..4463
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4095..4114)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4305..4326
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4489..4593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4594..4662,4663..5454)
                     /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(4812..4831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5625..6213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6114..6133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6501..6522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6537..6542,6543..6560,6561..6567)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6575..6591
                     /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     6580..6602
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6599..6615
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6599..6615
                     /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 gtatggtgag
     1021 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     1081 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1141 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1201 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     1261 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1321 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1381 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1441 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     1501 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1561 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1621 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1681 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gtggcggtgg
     1741 ctcgggcggt ggtgggtcga tggaagaagc tgagctggtg aagggaagac tccaggccat
     1801 cactgacaaa agaaagatac aggaagaaat ctcacagaag cgtctgaaaa tagaggaaga
     1861 aaaattgaaa caccaacatc tgaagaaaaa agccctgagg gagaaatggc tcctggatgg
     1921 aatcggcagt ggaaaagaac acgaagagat gaagaagcaa aatcagcaag accagcacca
     1981 gacccaggtt ctagagcaaa gcatcctcag gcttgagaaa gagatccaag atcttgaaaa
     2041 ggctgaactg caaatctcag ccaacgaaga ggcaattctg aagaagctaa agtcgatcga
     2101 gaagaccaca gaagacataa taagatccgt gaaggtggaa aaggaagaaa acccagaaga
     2161 gtcaatcgag gacatctatg ctaatatccc tgaccttcca agttcctaca taccttcccg
     2221 attaagaaag gaaagaaacg aaggaccaga tgacgaacaa aacagaaaag ctttgtatgc
     2281 catggaaatt aaagttgaaa aagatttgaa gactggagag agcgtagttc tgtcttcaat
     2341 accgctgcca tctgatgact ttaaaagcac agggataaaa gtgtatgagg accggcagaa
     2401 atcagtctat gctgttagct ccaatcagaa cacaacctac aatggcaccg acggcctcgc
     2461 acctgtcgaa gtggaggatc tcctaagaca agcctcagag aggaactcta aatcgcccac
     2521 agagtaccat gagccagtgt acgccaatcc gttttgcagg cctgtgaccc cacagagaga
     2581 aagagtcata agccctggac caaattttca agaaaggata atgatgaaaa cgaacggact
     2641 gggcaaccat gcaaatgaat ctgcgcacaa tatgaccgac gggctctcag agaggagaag
     2701 caacggtccc actcacacca gtcccactcg gccaacacct cagccccgat caatggttca
     2761 gcaagtggaa gagatggtcc acacccaaca aaagaggatg gcaagccctt gggaagaatc
     2821 cagcaacagg caaaacgaac acgaagtttc tccaaggatg gaactgagcc ccagcagagc
     2881 aagccctggg aaatcgggac cccagtgttc ttcacccact tgccaggagg agacagaaga
     2941 tgtcagatat aacatcgttc attccctgcc ttctgatgtg gatgacacag aacctgtaac
     3001 aatgattttc atggggtatc agcaggcaga tgataatgag gaagaaaaga agcttttgac
     3061 aggctacgac ggggtcatcc acgctgagct cgttgtgatt gatgacgagg cggaggataa
     3121 tgaaggacaa accgagaggc catcctacca tcccgtagct ccctacagtc aggtttacca
     3181 gccacccaaa ccaaccccac ttcctagaaa gagagcggaa gttaggccgt atgaaaacac
     3241 aaaccataaa tctcctcaca aaaattccat ctctctgaaa gagcaagagg aacgcttagg
     3301 cagccctgcc cggcactctc ccctggatgt cccagtagct ggagacggga ctgaggatcc
     3361 ttccttaaca gctttaagaa taagaatggc caagctgggg aaaaaggtga tctaataagc
     3421 tcgcgtggta cctctagagt cgacccgggc ggcctcgagg acggggtgaa ctacgcctga
     3481 ggatccgatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     3541 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     3601 tgtctctcac tcggaagcaa ttcgttgatc tgaatttcga ccacccataa tacccattac
     3661 cctggtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
     3721 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3781 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gccttaatta
     3841 acctaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3901 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3961 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     4021 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4081 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4141 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4201 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4261 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4321 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4381 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4441 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     4501 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4561 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4621 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4801 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5101 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5281 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5461 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5521 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5581 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5641 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5701 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5761 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5821 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5881 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5941 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6001 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6061 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6421 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6481 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6541 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6601 ggaaacagct atgaccatga ttacgccaga tttaattaag gccttaatta gg
//
