LOCUS       sequence-456468-        7308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7308)
  AUTHORS   Woo JS, Srikanth S, Nishi M, Ping P, Takeshima H, Gwack Y
  TITLE     Junctophilin-4, a component of the endoplasmic reticulum-plasma 
            membrane junctions, regulates Ca2+ dynamics in T cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Mar 8;113(10):2762-7. doi: 
            10.1073/pnas.1524229113. Epub 2016 Feb 29.
  PUBMED    26929330
REFERENCE   2  (bases 1 to 7308)
  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: 
            "10.1073/pnas.1524229113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-03-8- 8"; volume: "113"; issue: "10"; pages: "2762-7"
COMMENT     Sequence Label: p-mCherry-C1-mJP-4
FEATURES             Location/Qualifiers
     source          1..7308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        53..356
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        357..560
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     511..531
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             589..1296
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             1315..2022
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    4088..4209
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4125..4144)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4179..4198
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4216..4671)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4353..4374)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4565..4584
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4698..4802
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4804..5161
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4804..4995
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4827..4847)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5012..5147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5196..5990
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5196..5990
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5250..5269)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5860..5879
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6178..6197)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6222..6269
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6598..7186
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7087..7106
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
       61 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      121 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      181 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      241 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      301 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      361 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      421 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      481 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      541 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcat ggtgagcaag
      601 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
      661 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      721 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
      781 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
      841 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      901 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
      961 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     1021 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     1081 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     1141 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     1201 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     1261 cgccactcca ccggcggcat ggacgagctg tacaagtccg gactcagatc tagcatggtg
     1321 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1381 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1441 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1501 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1561 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1621 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1681 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1741 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     1801 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     1861 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     1921 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     1981 gagggccgcc actccaccgg cggcatggac gagctgtaca agtccggact cagatctcga
     2041 gcttcccccg ggggcaagtt cgactttgac gacgggggct gctacgtggg gggctgggag
     2101 gcggggcggg cacatggcta cggcgtgtgc accgggcccg gcgcccaggg agagtacagc
     2161 ggctgctggg ctcacggctt cgagtcgctg ggtgtcttca cggggcccgg cggacacagt
     2221 taccagggcc actggcagca gggcaagcgc gaagggctgg gcgtggagcg caagagccgc
     2281 tggacctacc gcggcgagtg gctgggcggg ctgaaggggc gcagcggcgt atgggagagc
     2341 gtgtccggcc tgcgctacgc cgggctctgg aaggacggct tccaggacgg ctacggcacc
     2401 gagacctact cggatggagg cacttaccaa ggccagtggc aagccgggaa acgccacggc
     2461 tacggggtgc gccaaagtgt accctaccac caggcggcgc ttctgcgctc accccgccgc
     2521 acctccctgg actccggcca cagcgacccc ccgacgccgc ctccacccct acccctacca
     2581 ggagatgaag gaggcagccc agcctccggc tcccgaggcg gcttcgtgct ggcggggcca
     2641 ggggacgccg acggggcgtc ttcccgtaag cgcaccccag cagcaggtgg attcttccgc
     2701 cgttcactgc tgctcagcgg gctccgggct ggtgggcgcc gcagctccct gggcagcaag
     2761 aggggatctc tacgcagcga ggtgagcagc gaagtgggca gtacaggacc cccgggctcc
     2821 gaggccagcg ggcccccaat cccagctcca cctgccctca tcgagggctc agccactgag
     2881 gtgtacgccg gtgagtggcg tgcggaccgg cgcagtggct atggagtgag ccagagatcc
     2941 aacgggctgc gctacgaagg cgagtggctg ggcaaccgac gccacggata cgggcgcacc
     3001 acccggcccg atggctcccg tgaggagggc aagtacaagc gtaaccggct ggtacacggg
     3061 ggacgtgtcc gcagcctcct gcctctggcc cttagacggg gcaaagtcaa ggagaaagtg
     3121 gacagggctg tcgaaggtgc ccgtcgagct gtgagtgctg cccgccagcg ccaggaaatt
     3181 gcggctgcca gggcagcaga cgccctccta aaagcagtgg cagccagcag cgtcgcggag
     3241 aaggctgtgg aggctgcgcg gatggccaag ctgatagccc aggatctaca acccatgtta
     3301 gaggccccag gccgcagacc cagacaggac tcaggaggtt ctgacacaga gcctttggat
     3361 gaggacagcc ctggggtata cgagaatgga ctgaccccct cagagggctc tccggaacta
     3421 cccagcagcc ccgcctcctc ccaccaacct tggcgagccc ctgcctgccg gagtccactg
     3481 cctcctggag ggaactgggg ccccttctcc agccctaaag cttggcccga ggagtggggg
     3541 ggtcctggtg agcaggcaga ggaactagct ggctatgagg ctgaggatga ggctggggtg
     3601 cagggtccag gaccaagaga tggttcccca cttctcggag gctgcagtga cagttcagga
     3661 agtcttcgag aggaggaggg ggaagacgaa gagtccttgc cccaactgag ggccccagga
     3721 ggctcagagt ctgagcctgt caccacgcca gttttgaggg gcctgtcttc aaggggtcct
     3781 gatgctgggt gcctgacaga agaggttgag gagcctgctc ccaccgagag gcctgcccag
     3841 ccgggagctg ccaaccccct ggtggtggga gccgtggcct tgctcgatct cagcctggcc
     3901 ttcctgttct cccagctcct cacctgagaa ttctgcagtc gacggtaccg cgggcccggg
     3961 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     4021 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4081 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4141 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4201 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4261 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4321 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4381 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4441 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4501 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4561 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4621 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4681 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4741 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4801 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4861 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4921 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4981 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5041 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5101 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5161 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5221 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5281 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5341 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     5401 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5461 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5521 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5581 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5641 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5701 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     5761 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5821 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     5881 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     5941 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6001 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6061 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6121 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     6181 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     6241 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     6301 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     6361 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     6421 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     6481 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6541 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6601 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6661 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6721 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6781 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6841 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6901 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6961 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7021 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7081 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7141 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7201 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7261 atcccctgat tctgtggata accgtattac cgccatgcat tagttatt
//
