LOCUS       sequence-349875-        6066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NKD2-mCherry fusion protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6066)
  AUTHORS   Wu B, Woo JS, Vila P, Jew M, Leung J, Sun Z, Srikanth S, Gwack Y
  TITLE     NKD2 mediates stimulation-dependent ORAI1 trafficking to augment 
            Ca(2+) entry in T cells.
  JOURNAL   Cell Rep. 2021 Aug 24;36(8):109603. doi: 
            10.1016/j.celrep.2021.109603.
  PUBMED    34433025
REFERENCE   2  (bases 1 to 6066)
  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: "Cell Rep.";
            date: "2021-08-24"; volume: "36(8):109603. doi"; pages: " 
            10.1016/j.celrep.2021.109603"
COMMENT     Sequence Label: pN1-mCherry-NKD2
FEATURES             Location/Qualifiers
     source          1..6066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        81..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1953..1979
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     regulatory      2037..2046
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2043..2753
                     /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"
     primer_bind     complement(2188..2206)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2456..2475
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2874..2995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2911..2930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2965..2984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3002..3457)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3139..3160)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3351..3370
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3484..3588
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3590..3947
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3590..3781
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3613..3633)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3798..3933
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3982..4776
                     /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             3982..4776
                     /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(4036..4055)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4646..4665
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4964..4983)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5008..5055
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5384..5972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5873..5892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
      601 cgtcagatcc gctagcgcta ccggactcag atctcgagat ggggaaactg cagtcgaagc
      661 acgccgccgc cgcccgcaag cggagagaga gcccggaagg ggacagcttc gtggcgtccg
      721 cgtacgcgag cggccgcaaa ggcgcggagg aagcggagcg gcgcgcgcgg gacaagcagg
      781 agctgcccaa tggggacccc aaggaggggc ctttccggga ggaccagtgt cccctacagg
      841 tggcactccc cgctgagaaa gctgagggcc gcgagcaccc gggacaactc ctcagcgcag
      901 atgacggaga gagggcagca aaccgcgagg gcccgcgagg accgggcggg cagcgcctca
      961 acattgacgc actccagtgc gatgtctcgg tggaggagga cgaccgccag gagtggacgt
     1021 tcacgctcta tgactttgac aactgcggga aggtcaccag ggaggacatg tccagcctca
     1081 tgcacaccat ctatgaggtc gtggatgcct cggtcaacca ctcctcgggc agcagcaaga
     1141 ccctccgtgt gaagctaacc gtcagccctg agccctccag caagaggaag gagggtcctc
     1201 ctgctggcca ggaccgggag cccacccgtt gcaggatgga gggtgaactg gcagaggagc
     1261 caagggtggc tgacaggagg ttgtctgcac acgtcaggag gcccagtact gacccccagc
     1321 cctgctcgga gcgggggccc tactgcgtgg acgagaacac ggagcgcaga aaccactacc
     1381 tggacctcgc cgggattgag aactacacgt ccagattcgg ccctgggtcc cctcctgtgc
     1441 aagcaaagca ggagccccag ggcagggcct cgcacctcca ggcccggtcc cgctcccagg
     1501 agccagatac acatgccgta caccaccgca ggtcacaggt gctggtggaa cacgtcgtgc
     1561 cagcctcgga gcctgctgcc cgggccctgg acacgcagcc ccggccgaag gggccggaga
     1621 agcagttcct caagtccccc aagggctccg ggaagccgcc tggggtgcca gccagcagca
     1681 agtccgggaa agccttcagc tactacctgc cggccgtcct gccgccccag gcccctcagg
     1741 acggccacca cctcccgcag cccccaccgc caccctacgg ccacaagcgg taccgccaaa
     1801 agggcaggga gggccactcg ccactcaagg ccccacacgc tcagcctgcc acagtggagc
     1861 acgaggtggt gcgggacctg ccgcccacgc cagcaggaga gggctacgcg gtgccagtga
     1921 tccagcggca cgagcaccac caccaccacg agcaccacca ccaccaccac caccaccact
     1981 tccacccgtc ccgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     2041 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     2101 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     2161 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     2221 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     2281 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     2341 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2401 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2461 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2521 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2581 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2641 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2701 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tagcggccgc
     2761 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2821 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2881 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2941 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     3001 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3061 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3121 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3181 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3241 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3301 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3361 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3421 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3601 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3661 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3721 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3781 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3841 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3901 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3961 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4021 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4081 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4141 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4201 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4261 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4321 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4381 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4441 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4501 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4561 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4621 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4681 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4741 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4801 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4861 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4921 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4981 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5041 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5101 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5161 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5221 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5281 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5341 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5401 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5461 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5521 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5581 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5641 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5701 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5761 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5821 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5881 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5941 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6001 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6061 tggata
//
