LOCUS       sequence-429373-        6697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expressing Falg tag and the coding sequence of mouse Nexn.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6697)
  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 6697)
  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: pCMV-3*flag-NEXN
FEATURES             Location/Qualifiers
     source          1..6697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..97
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     89..106
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     90..106
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        267..646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        647..850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     801..821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             880..945
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2987..3609
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3186..3205)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3633..3653)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3638..3967
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3818..3953
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4005..4024)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4006..4024)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4038..4054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4038..4054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4051..4073)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4062..4078
                     /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(4086..4092,4093..4110,4111..4116))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4131..4152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4269..4286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4440..5028)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4520..4539)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5199..5990,5991..6059))
                     /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     5822..5841
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6060..6164)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6191..6646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6278..6297)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6488..6509
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
       61 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctgatcta
      121 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa
      181 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
      241 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc
      301 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      361 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      421 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      481 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
      541 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
      601 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      661 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      721 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      781 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      841 aagcagagct cgtttagtga accgtcagaa ttaaccatgg actacaaaga ccatgacggt
      901 gattataaag atcatgacat cgattacaag gatgacgatg acaagcttgc ggccgcgaat
      961 tcgatgaatg acgtttcgca aaaggcagag attctgcttt cctcatgtaa acctgtccct
     1021 aagtcctatg tgccaaagct tggcaaggga gatgtaaagg acaagtttga agccatgcag
     1081 agggccaggg aagaaagaaa tcaaaggcga tctagagacg aaaagcaaag aagaaaagaa
     1141 caatatatta gagagagaga atggaacagg agaaagcagg agattaaaga aatgcttgct
     1201 tctgatgatg aggaagaaag ttctcctaaa atagaaaagg cttatgtccc aaagctaaca
     1261 gggacagtta aaggtaaatt cgatgaaatg gagaaacaca gacaggaaga acaaagaaag
     1321 agaacagaag aagaaagaaa acgcagaatt gagcaggatc tgttagaaaa gaggaaaata
     1381 cagcgcgaat tagcaaaaag agcggagcag attgaggaca taaacaatac gggaaccgag
     1441 tccgcatcag aggaaggaga tgactcacta cttataacag tggtacctgc caaatcatac
     1501 aaaacacctg gaaaaacaaa ggatcctgag gacctggaca gagaagaagg aaatgggaga
     1561 accaaccatg aagaagataa aatgagatat gaagaagagt gccgagttct caaagaagca
     1621 aaatgtcttt cattagtcat ggatgatgaa acagaggcca aaaaagaatc acattttcct
     1681 ggaaaattga aatcaacttt tgaagaactg gaacgacaaa gacaagaaaa tcgcaagaag
     1741 caagcggaag aggaagcgag acgacgctta gaagaagaga gacgttcctt tgaggaggcg
     1801 agaaggcaca tggtaaatga ggaggatgaa aaccaagata gagaaacagt ttttaaagag
     1861 taccgcccag gaaaactcaa actcagtttt gaggaaatag aaaggcaaag gagagaagac
     1921 gaaaaacgaa aagcagagga ggaggcccga aggagaatag aggaagagaa ggcggcattt
     1981 gctgaagcaa ggagaagcat ggtattagat gatgattccc cagagattta taaaacagtg
     2041 tctcaagaat ctcttactcc tggaaaacta gaaattaatt ttgaacaatt attaagacaa
     2101 aaaatggaag aagaaagacg acgaacagag gaagaacgca ggcataaact agaaatggag
     2161 aaacaggaat ttgaacaact gagacaagaa atgggaaagg aagaagaaga aaatgaaagc
     2221 tttgggttga gtagagaata tgaagaatta attaaattaa aaaggagtgg ctcaattcaa
     2281 gctaaaaatc taaaaagcaa gtttgaaaaa attggacaat tgtctgaaaa agaagtacag
     2341 aaaaagatag aagaagagcg agcaaaaagg cgagcaattg accttgaaat caaagagcga
     2401 gaagcagaaa acttccatga ggacgatgat gttgatgtaa ggcctgcaaa gaagagtgag
     2461 tctcctttta ctcataaggt gaacatgaaa gccaggtttg aacaaatggc aaaggcaaga
     2521 gaagaggaag aacaaagaag aattgaagaa caaaagttac tacgaatgca gtttgaacag
     2581 aaagaaatcg atgcggcatt acagaagaaa agagaagatg aggaggaaga agaaggtagc
     2641 attgttaatg gctccactac tgaagatgaa gagcaaacca gatcaggagc tccatggttc
     2701 aaaaagcctt taagaaatac ctcagttgta gacagtgagc cagtcagatt tactgttaaa
     2761 gtaacaggag aacccaagcc ggaaattaca tggtggtttg aaggagaaat actgcaggat
     2821 ggagaagact atcagtacat cgaaagaggt gaaacttact gcctgtattt accggaaacc
     2881 ttcccagaag atggaggaga gtacatgtgt aaggcagtca acaataaagg ctcagcagcg
     2941 agcacctgca ttcttaccat tgaaatggat gactactagg gatcccgggt ggcatccctg
     3001 tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct
     3061 tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat
     3121 ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg
     3181 cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc
     3241 cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc
     3301 atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt
     3361 ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt
     3421 gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt taaaataact
     3481 ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc aaccggtggg
     3541 acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc agtagatgcc
     3601 tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag ggtgtggaaa
     3661 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3721 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3781 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3841 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3901 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3961 ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat agtgagtcgt
     4021 attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4081 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4141 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4201 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4261 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4321 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4381 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4441 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4501 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4561 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4621 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4681 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4741 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4801 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4861 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4921 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4981 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5041 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5101 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5161 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5221 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5281 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5341 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5401 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5461 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5521 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5581 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5641 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5701 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5761 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5821 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5881 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5941 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6001 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6061 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6121 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6181 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6241 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6301 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6361 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6421 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     6481 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     6541 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     6601 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca
     6661 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggc
//
