LOCUS       sequence-250350-        6369 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6369)
  AUTHORS   Gwack Y, Srikanth S, Feske S, Cruz-Guilloty F, Oh-hora M, Neems DS, 
            Hogan PG, Rao A
  TITLE     Biochemical and functional characterization of Orai proteins.
  JOURNAL   J Biol Chem. 2007 Jun 1. 282(22):16232-43.
  PUBMED    17293345
REFERENCE   2  (bases 1 to 6369)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2007 Jun 1. 282(22):16232-43."
COMMENT     Sequence Label: pcDNA3.1 Orai1
FEATURES             Location/Qualifiers
     source          1..6369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     527..546
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        621..725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(726..794,795..1586)
                     /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     complement(944..963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1757..2345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2246..2265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2499..2516
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2633..2654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2669..2674,2675..2692,2693..2699)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2707..2723
                     /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)."
     primer_bind     2712..2734
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2731..2747
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2731..2747
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    2796..2917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2807..2826)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2861..2880
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3093..3887)
                     /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(3204..3223)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3814..3833
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3954..4283)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3968..4103
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4022..4041)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4268..4288
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4297..4725)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4407..4428)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4619..4638
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4771..4995
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4984..5001
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5024..5041)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(5057..5086)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      5703..5712
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6077..6096)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6078..6096)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6141..6344)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6170..6190)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(6345..6369,1..355)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
       61 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
      121 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
      181 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
      241 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
      301 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcaacgc
      361 gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc
      421 atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
      481 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
      541 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
      601 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
      661 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      721 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      781 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      841 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      901 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      961 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1021 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1081 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1141 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1201 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1261 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1321 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1381 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1441 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1501 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1561 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1621 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1681 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1741 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1801 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1861 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1921 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1981 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2041 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2101 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2161 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2221 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2281 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2341 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2401 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2461 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2521 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     2581 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2641 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2701 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2761 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     2821 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2881 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     2941 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     3001 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     3061 ggtcggtcat ttcgcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     3121 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     3181 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     3241 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     3301 atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc
     3361 gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     3421 tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct
     3481 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     3541 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     3601 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     3661 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     3721 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     3781 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     3841 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct
     3901 catcctgtct cttgatcaga tccgaaaatg gatatacaag ctcccgggag ctttttgcaa
     3961 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggcagaggcg
     4021 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     4081 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     4141 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
     4201 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
     4261 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
     4321 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     4381 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     4441 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     4501 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     4561 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     4621 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     4681 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga taccccctag
     4741 agccccagct ggttctttcc gcctcagaag ccatagagcc caccgcatcc ccagcatgcc
     4801 tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
     4861 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
     4921 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
     4981 aactagaagg cacagtcgag gctgatcagc gggtttaaac tcaatggtga tggtgatgat
     5041 gaccggtatg catattcaga tcctcttctg agatgagttt ttgttcgaag ggccctctag
     5101 actcgagggc atagtggctg ccgggcgtca gggggtggtc ccctctgtgg tccagctggt
     5161 cctgtaagcg ggcaaactcc gccagctcgt tgagctcctg gaactgtcgg tcggtcttat
     5221 ggctaaccag tgagcggtag aagtggacgg cgaagacgat aaagatcagg ccgaagggca
     5281 ccatgatggt ggtcgaggcg atggcagctg cctggcccgg ggtgatgccg ctggtgctga
     5341 cgttggctgc tgcgccactg gcggggggct tgctggtggg ccttggctgg cctggctgct
     5401 tcttgagggg caagaacttg acccagcaga gcagcaccac ctcagctagg aagagcagcg
     5461 tgccaatgac ggtggagaag gcccaggcca gctcgatgtg gcggtgcatg cgctcatggg
     5521 gggactcctt gaccgagttg agattgtgca cgttgctcac cgcctcgatg ttgggcagga
     5581 tgcaggtgct gatcatgagc gcaaacaggt gcacagccac cagcactgtg gtgcaggcac
     5641 tgaaggcgat gagcagcccc ggtgggtagt cgtggtcagc gtccagctgc acctccacca
     5701 ttgccaccat ggcgaagccg gagagcagag ccgaggtccg gctggaggct ttaagcttgg
     5761 cgcggctcaa gtagagcttg cgccaggaca gcgcctgcat ggagtgctcg ttgaggccca
     5821 tcacctcgga gtaactctgg ccgatccagt ccgggtaggt gacggcggac ggcggcggtg
     5881 gcggggcccc cgggggctcc ccgtccccgc tgcggcggcg gctccggcgg ctgccgctgg
     5941 tggtgctgcc gccgcttggg ggaagctcgg gactgctacg gctcgggggc ggcgcgggct
     6001 ccggatgcat gaattccacc acactggact agtggatccg agctcggtac caagcttaac
     6061 tagccagctt gggtctccct atagtgagtc gtattaattt cgataagcca gtaagcagtg
     6121 ggttctctag ttagccagag agctctgctt atatagacct cccaccgtac acgcctaccg
     6181 cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg
     6241 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
     6301 aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg
     6361 actaatacg
//
