LOCUS       sequence-349873-        6066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes human NKD2 G2A (2nd Gly residue to Ala) mutant cDNA with a 
            C-terminal mCherry fusion..
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: pN1mcherry-NKD2G2A
FEATURES             Location/Qualifiers
     source          1..6066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        123..426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        427..630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     581..601
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1995..2021
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     regulatory      2079..2088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2085..2795
                     /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(2230..2248)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2498..2517
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2916..3037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2953..2972)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3007..3026
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3044..3499)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3181..3202)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3393..3412
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3526..3630
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3632..3989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3632..3823
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3655..3675)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3840..3975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4024..4818
                     /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             4024..4818
                     /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(4078..4097)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4688..4707
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5006..5025)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5050..5097
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5426..6014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5915..5934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
       61 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      121 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      601 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      661 taccggactc agatctcgag atggcgaaac tgcagtcgaa gcacgccgcc gccgcccgca
      721 agcggagaga gagcccggaa ggggacagct tcgtggcgtc cgcgtacgcg agcggccgca
      781 aaggcgcgga ggaagcggag cggcgcgcgc gggacaagca ggagctgccc aatggggacc
      841 ccaaggaggg gcctttccgg gaggaccagt gtcccctaca ggtggcactc cccgctgaga
      901 aagctgaggg ccgcgagcac ccgggacaac tcctcagcgc agatgacgga gagagggcag
      961 caaaccgcga gggcccgcga ggaccgggcg ggcagcgcct caacattgac gcactccagt
     1021 gcgatgtctc ggtggaggag gacgaccgcc aggagtggac gttcacgctc tatgactttg
     1081 acaactgcgg gaaggtcacc agggaggaca tgtccagcct catgcacacc atctatgagg
     1141 tcgtggatgc ctcggtcaac cactcctcgg gcagcagcaa gaccctccgt gtgaagctaa
     1201 ccgtcagccc tgagccctcc agcaagagga aggagggtcc tcctgctggc caggaccggg
     1261 agcccacccg ttgcaggatg gagggtgaac tggcagagga gccaagggtg gctgacagga
     1321 ggttgtctgc acacgtcagg aggcccagta ctgaccccca gccctgctcg gagcgggggc
     1381 cctactgcgt ggacgagaac acggagcgca gaaaccacta cctggacctc gccgggattg
     1441 agaactacac gtccagattc ggccctgggt cccctcctgt gcaagcaaag caggagcccc
     1501 agggcagggc ctcgcacctc caggcccggt cccgctccca ggagccagat acacatgccg
     1561 tacaccaccg caggtcacag gtgctggtgg aacacgtcgt gccagcctcg gagcctgctg
     1621 cccgggccct ggacacgcag ccccggccga aggggccgga gaagcagttc ctcaagtccc
     1681 ccaagggctc cgggaagccg cctggggtgc cagccagcag caagtccggg aaagccttca
     1741 gctactacct gccggccgtc ctgccgcccc aggcccctca ggacggccac cacctcccgc
     1801 agcccccacc gccaccctac ggccacaagc ggtaccgcca aaagggcagg gagggccact
     1861 cgccactcaa ggccccacac gctcagcctg ccacagtgga gcacgaggtg gtgcgggacc
     1921 tgccgcccac gccagcagga gagggctacg cggtgccagt gatccagcgg cacgagcacc
     1981 accaccacca cgagcaccac caccaccacc accaccacca cttccacccg tcccgaattc
     2041 tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc caccatggtg agcaagggcg
     2101 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     2161 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     2221 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     2281 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     2341 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2401 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     2461 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     2521 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     2581 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     2641 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     2701 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2761 actccaccgg cggcatggac gagctgtaca agtagcggcc gcgactctag atcataatca
     2821 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2881 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2941 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3001 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     3061 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3121 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3181 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3241 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3301 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3361 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3421 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3481 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3541 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3601 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3661 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3721 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3781 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3841 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3901 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3961 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4021 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4081 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4141 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4201 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4261 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4321 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4381 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4441 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4501 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4561 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4621 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4681 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4741 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4801 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4861 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4921 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4981 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5041 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5101 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5161 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5221 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5281 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5341 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5401 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5461 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5521 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5581 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5641 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5701 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5761 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5821 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5881 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5941 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6001 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6061 cttttg
//
