LOCUS       sequence-429372-        7107 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expressing HA tag and the coding sequence of mouse Palmd.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7107)
  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 7107)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pCMV-3*HA-Palmd
FEATURES             Location/Qualifiers
     source          1..7107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(304..1095,1096..1164))
                     /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     927..946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1165..1269)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1344..1363)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1535..1914
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1915..2118
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2069..2089
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2163..2182
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2163..2181
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2199..2208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2214..2303
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(3998..4015)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4004..4228
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4274..4702
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4361..4380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4571..4592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4711..4731)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4716..5045
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4896..5031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4958..4977
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5112..5906
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5166..5185)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5776..5795
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6080..6201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6117..6136)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6171..6190
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6250..6266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6250..6266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6263..6285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6274..6290
                     /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)."
     promoter        complement(join(6298..6304,6305..6322,6323..6328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6343..6364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6481..6498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6652..7107,1..133))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6732..6751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
       61 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
      121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
      181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
      241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
      301 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
      361 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
      421 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
      481 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
      541 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
      601 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
      661 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
      721 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
      781 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
      841 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
      901 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
      961 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     1021 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     1081 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     1141 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     1201 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1261 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     1321 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     1381 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     1441 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     1501 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     1561 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     1621 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     1681 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     1741 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     1801 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     1861 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     1921 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     1981 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     2041 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     2101 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     2161 attaatacga ctcactatag ggagacccaa gctggctagc caccatggcc gtttacccat
     2221 acgatgttcc tgactatgcg ggctatccct atgacgtccc ggactatgca ggatcctatc
     2281 catatgacgt tccagattac gctaccggta tggaagaagc tgagctggtg aagggaagac
     2341 tccaggccat cactgacaaa agaaagatac aggaagaaat ctcacagaag cgtctgaaaa
     2401 tagaggaaga aaaattgaaa caccaacatc tgaagaaaaa agccctgagg gagaaatggc
     2461 tcctggatgg aatcggcagt ggaaaagaac acgaagagat gaagaagcaa aatcagcaag
     2521 accagcacca gacccaggtt ctagagcaaa gcatcctcag gcttgagaaa gagatccaag
     2581 atcttgaaaa ggctgaactg caaatctcag ccaacgaaga ggcaattctg aagaagctaa
     2641 agtcgatcga gaagaccaca gaagacataa taagatccgt gaaggtggaa aaggaagaaa
     2701 acccagaaga gtcaatcgag gacatctatg ctaatatccc tgaccttcca agttcctaca
     2761 taccttcccg attaagaaag gaaagaaacg aaggaccaga tgacgaacaa aacagaaaag
     2821 ctttgtatgc catggaaatt aaagttgaaa aagatttgaa gactggagag agcgtagttc
     2881 tgtcttcaat accgctgcca tctgatgact ttaaaagcac agggataaaa gtgtatgagg
     2941 accggcagaa atcagtctat gctgttagct ccaatcagaa cacaacctac aatggcaccg
     3001 acggcctcgc acctgtcgaa gtggaggatc tcctaagaca agcctcagag aggaactcta
     3061 aatcgcccac agagtaccat gagccagtgt acgccaatcc gttttgcagg cctgtgaccc
     3121 cacagagaga aagagtcata agccctggac caaattttca agaaaggata atgatgaaaa
     3181 cgaacggact gggcaaccat gcaaatgaat ctgcgcacaa tatgaccgac gggctctcag
     3241 agaggagaag caacggtccc actcacacca gtcccactcg gccaacacct cagccccgat
     3301 caatggttca gcaagtggaa gagatggtcc acacccaaca aaagaggatg gcaagccctt
     3361 gggaagaatc cagcaacagg caaaacgaac acgaagtttc tccaaggatg gaactgagcc
     3421 ccagcagagc aagccctggg aaatcgggac cccagtgttc ttcacccact tgccaggagg
     3481 agacagaaga tgtcagatat aacatcgttc attccctgcc ttctgatgtg gatgacacag
     3541 aacctgtaac aatgattttc atggggtatc agcaggcaga tgataatgag gaagaaaaga
     3601 agcttttgac aggctacgac ggggtcatcc acgctgagct cgttgtgatt gatgacgagg
     3661 cggaggataa tgaaggacaa accgagaggc catcctacca tcccgtagct ccctacagtc
     3721 aggtttacca gccacccaaa ccaaccccac ttcctagaaa gagagcggaa gttaggccgt
     3781 atgaaaacac aaaccataaa tctcctcaca aaaattccat ctctctgaaa gagcaagagg
     3841 aacgcttagg cagccctgcc cggcactctc ccctggatgt cccagtagct ggagacggga
     3901 ctgaggatcc ttccttaaca gctttaagaa taagaatggc caagctgggg aaaaaggtga
     3961 tctagggtac caagcttaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc
     4021 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4081 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4141 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4201 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     4261 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4321 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4381 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4441 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4501 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4561 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4621 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4681 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4741 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4801 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4861 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4921 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4981 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5041 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     5101 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5161 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5221 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5281 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5341 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5401 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5461 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5521 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5581 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5641 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5701 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5761 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5821 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5881 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5941 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     6001 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6061 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     6121 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     6181 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     6241 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6301 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     6361 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6421 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6481 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6541 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6601 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6661 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6721 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6781 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6841 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6901 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6961 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     7021 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     7081 ctacactaga agaacagtat ttggtat
//
