LOCUS       sequence-171022-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal Venus (YFP) fragment fused to full-length ORAI1; for 
            BiFC..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  AUTHORS   Hertel F, Mo GC, Duwe S, Dedecker P, Zhang J
  TITLE     RefSOFI for Mapping Nanoscale Organization of Protein-Protein 
            Interactions in Living Cells.
  JOURNAL   Cell Rep. 2016 Jan 12;14(2):390-400. doi: 
            10.1016/j.celrep.2015.12.036. Epub 2015 Dec 31.
  PUBMED    26748717
REFERENCE   2  (bases 1 to 6972)
  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: 
            "10.1016/j.celrep.2015.12.036"; journalName: "Cell Rep"; date: 
            "2016-01-12- 12"; volume: "14"; issue: "2"; pages: "390-400"
COMMENT     Sequence Label: pcDNA3-Venus-173-N-ORAI1
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             922..939
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             943..975
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(979..987,988..1002)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      1009..1018
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1015..1017,1018..1020,1021..1530)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNI"
     primer_bind     complement(1060..1081)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1321..1340)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      1856..1865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2524..2542)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2525..2542)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2562..2579)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2568..2792
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2838..3266
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2925..2944)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3135..3156
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3275..3295)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3280..3610
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3461..3596
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3523..3542
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3677..4471
                     /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(3731..3750)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4341..4360
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4645..4766
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4682..4701)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4736..4755
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4815..4831)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4815..4831)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4828..4850)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4839..4855
                     /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(4863..4869,4870..4887,4888..4893))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4908..4929
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5046..5063)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5217..5805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5297..5316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5976..6767,6768..6836))
                     /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     6599..6618
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6837..6941)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc
      901 gccaccatgg tgcggggttc tcatcatcat catcatcatg gtatggctag catgactggt
      961 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatcccgc caccatggtg
     1021 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1081 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1141 ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1201 accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac
     1261 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1321 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1381 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1441 gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     1501 aaggccaact tcaagatccg ccacaacatc ggaggtagcg gaggtggaag tggtaccatg
     1561 catccggagc ccgccccgcc cccgagccgt agcagtcccg agcttccccc aagcggcggc
     1621 agcaccacca gcggcagccg ccggagccgc cgccgcagcg gggacgggga gcccccgggg
     1681 gccccgccac cgccgccgtc cgccgtcacc tacccggact ggatcggcca gagttactcc
     1741 gaggtgatga gcctcaacga gcactccatg caggcgctgt cctggcgcaa gctctacttg
     1801 agccgcgcca agcttaaagc ctccagccgg acctcggctc tgctctccgg cttcgccatg
     1861 gtggcaatgg tggaggtgca gctggacgct gaccacgact acccaccggg gctgctcatc
     1921 gccttcagtg cctgcaccac agtgctggtg gctgtgcacc tgtttgcgct catgatcagc
     1981 acctgcatcc tgcccaacat cgaggcggtg agcaacgtac acaatctcaa ctcggtcaag
     2041 gagtcccccc atgagcgcat gcaccgccac atcgagctgg cctgggcctt ctccaccgtc
     2101 attggcacgc tgctcttcct agctgaggtg gtgctgctct gctgggtcaa gttcttgccc
     2161 ctcaagaagc agccaggcca gccaaggccc accagcaagc cccccgccag tggcgcagca
     2221 gccaacgtca gcaccagcgg catcaccccg ggccaggcag ctgccatcgc ctcgaccacc
     2281 atcatggtgc ccttcggcct gatctttatc gtcttcgccg tccacttcta ccgctcactg
     2341 gttagccata agaccgaccg acagttccag gagctcaacg agctggcgga gtttgcccgc
     2401 ttacaggacc agctggacca cagaggggac caccccctga cgcccggcag ccactatgcc
     2461 taagaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
     2521 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     2581 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2641 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2701 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2761 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2821 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2881 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2941 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3001 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3061 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3121 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3181 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3241 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3301 aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca attagtcagc
     3361 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3421 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3481 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3541 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3601 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     3661 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3721 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3781 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3841 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3901 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3961 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4021 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4081 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4141 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4201 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4261 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4321 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4381 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4441 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4501 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4561 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4621 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4681 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4741 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4801 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4861 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4921 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4981 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5041 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5101 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5161 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5221 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5281 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5341 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5401 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5461 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5521 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5581 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5641 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5701 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5761 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5821 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5881 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5941 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6001 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6061 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6121 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6181 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6241 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6301 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6361 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6421 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6481 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6541 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6601 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6661 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6721 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6781 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6841 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6901 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6961 ccacctgacg tc
//
