LOCUS       Exported                6971 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  N-terminal Venus (YFP) fragment fused to full-length ORAI1; for 
            BiFC..
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3-Venus-173-N-ORAI1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  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 6971)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6971
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             22..54
                     /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             58..81
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      88..97
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             94..609
                     /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(139..160)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(400..419)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     regulatory      935..944
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(1603..1621)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1604..1621)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1641..1658)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1647..1871
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1917..2345
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2004..2023)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2214..2235
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2354..2374)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2359..2688
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2539..2674
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2601..2620
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2755..3549
                     /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(2809..2828)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3419..3438
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3723..3844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3760..3779)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3814..3833
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3893..3909)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3893..3909)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3906..3928)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3917..3933
                     /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(3941..3971)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3986..4007
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4124..4141)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4295..4883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4375..4394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5054..5914)
                     /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     5677..5696
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5915..6019)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6094..6113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6285..6664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6665..6868
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6819..6839
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6913..6932
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6913..6931
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6951..6960
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
ORIGIN
        1 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
       61 ctgtacgacg atgacgataa ggatcccgcc accatggtga gcaagggcga ggagctgttc
      121 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      181 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
      241 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg
      301 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      361 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      421 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      481 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
      541 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
      601 cacaacatcg gaggtagcgg aggtggaagt ggtaccatgc atccggagcc cgccccgccc
      661 ccgagccgta gcagtcccga gcttccccca agcggcggca gcaccaccag cggcagccgc
      721 cggagccgcc gccgcagcgg ggacggggag cccccggggg ccccgccacc gccgccgtcc
      781 gccgtcacct acccggactg gatcggccag agttactccg aggtgatgag cctcaacgag
      841 cactccatgc aggcgctgtc ctggcgcaag ctctacttga gccgcgccaa gcttaaagcc
      901 tccagccgga cctcggctct gctctccggc ttcgccatgg tggcaatggt ggaggtgcag
      961 ctggacgctg accacgacta cccaccgggg ctgctcatcg ccttcagtgc ctgcaccaca
     1021 gtgctggtgg ctgtgcacct gtttgcgctc atgatcagca cctgcatcct gcccaacatc
     1081 gaggcggtga gcaacgtaca caatctcaac tcggtcaagg agtcccccca tgagcgcatg
     1141 caccgccaca tcgagctggc ctgggccttc tccaccgtca ttggcacgct gctcttccta
     1201 gctgaggtgg tgctgctctg ctgggtcaag ttcttgcccc tcaagaagca gccaggccag
     1261 ccaaggccca ccagcaagcc ccccgccagt ggcgcagcag ccaacgtcag caccagcggc
     1321 atcaccccgg gccaggcagc tgccatcgcc tcgaccacca tcatggtgcc cttcggcctg
     1381 atctttatcg tcttcgccgt ccacttctac cgctcactgg ttagccataa gaccgaccga
     1441 cagttccagg agctcaacga gctggcggag tttgcccgct tacaggacca gctggaccac
     1501 agaggggacc accccctgac gcccggcagc cactatgcct aagaattctg cagatatcca
     1561 tcacactggc ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa
     1621 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1681 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1741 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1801 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1861 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1921 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1981 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2041 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2101 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2161 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2221 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2281 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2341 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2401 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2461 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2521 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2581 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2641 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2701 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2761 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2821 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2881 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     2941 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3001 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3061 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3121 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3181 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3241 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3301 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3361 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3421 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3481 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3541 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3601 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3661 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3721 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3781 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3841 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     3901 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3961 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4021 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4081 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4141 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4201 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4261 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4321 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4381 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4441 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4501 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4561 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4621 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4681 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4741 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4801 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4861 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4921 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4981 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5041 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5101 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5161 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5221 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5281 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5341 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5401 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5461 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5521 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5581 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5641 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5701 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5761 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5821 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5881 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5941 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6001 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     6061 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6121 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     6181 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     6241 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     6301 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     6361 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     6421 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     6481 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     6541 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     6601 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     6661 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     6721 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     6781 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     6841 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
     6901 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgcggcc gccaccatgg
     6961 tgcggggttc t
//
