LOCUS       sequence-421295-        6228 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6228)
  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 6228)
  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: "J Biol 
            Chem. 2007 Jun 1. 282(22):16232-43."
COMMENT     Sequence Label: pcDNA3.1 Orai2
FEATURES             Location/Qualifiers
     source          1..6228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(107..127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        112..441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      292..427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     354..373
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             508..1302
                     /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(562..581)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1172..1191
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1478..1599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1515..1534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1569..1588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1648..1664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1648..1664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1661..1683)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1672..1688
                     /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(1696..1702,1703..1720,1721..1726))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1741..1762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1879..1896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2050..2638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2130..2149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2809..3600,3601..3669))
                     /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     3432..3451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3670..3774)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3849..3868)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4040..4419
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4420..4623
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4574..4594
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4668..4687
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4668..4686
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4974..4983
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5537..5566
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5582..5599
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5622..5639)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5628..5852
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5898..6228,1..98)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5985..6004)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6195..6216
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
       61 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
      121 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
      181 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      241 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
      301 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
      361 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
      421 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
      481 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
      541 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
      601 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
      661 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
      721 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
      781 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
      841 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
      901 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
      961 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     1021 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     1081 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     1141 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     1201 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     1261 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     1321 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     1381 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     1441 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     1501 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1561 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     1621 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     1681 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1741 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1801 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1861 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1921 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1981 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2041 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2101 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2161 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2221 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2281 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2341 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2401 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2461 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2521 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2581 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2641 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2701 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2761 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2821 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2881 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2941 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3001 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3061 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3121 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3181 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3241 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3301 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3361 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3421 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3481 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3541 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3601 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3661 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3721 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3781 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     3841 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     3901 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     3961 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     4021 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     4081 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     4141 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     4201 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     4261 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     4321 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     4381 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     4441 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     4501 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     4561 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     4621 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
     4681 tatagggaga cccaagctgg ctagttaagc ttggtaccga gctcggatcc actagtccag
     4741 tgtggtggaa ttcatgagtg ctgagcttaa cgtgcctatc gacccctctg ctcctgcctg
     4801 ccctgagcca ggccataagg gcatggatta ccgggactgg gtccgccgca gctacctgga
     4861 actggtcacc tctaaccacc actcggtaca ggccctgtcg tggcggaagc tctacctgag
     4921 cagggccaag ctgaaggcct ccagcaggac ctccgccctc ctctccggct ttgccatggt
     4981 ggccatggtg gaggtgcagc tggagacgca gtaccagtac ccgcggccgc tgctgattgc
     5041 cttcagcgcc tgcaccacgg tgctggtggc cgtgcacctg ttcgccctcc tcatcagcac
     5101 ctgcatcctg cccaatgtgg aggccgtgag caacatccac aacctgaact ccatcagcga
     5161 gtccccgcat gagcgcatgc acccctacat cgagctggcc tggggcttct ccaccgtgct
     5221 tggcatccta ctcttcctgg ccgaggtggt gctgctctgc tggatcaagt tcctccccgt
     5281 ggatgcccgg cgccagcctg gccccccacc tggccctggg agtcacacgg gctggcaggc
     5341 cgccctggtg tccaccatca tcatggtgcc cgtgggcctc atcttcgtgg tcttcaccat
     5401 ccacttctac cgctccctgg tgcgccacaa aacggagcgc cacaaccgcg agatcgagga
     5461 gctccacaag ctcaaggtcc agctggacgg gcatgagcgc agcctgcagg tcttgctcga
     5521 gtctagaggg cccttcgaac aaaaactcat ctcagaagag gatctgaata tgcataccgg
     5581 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag
     5641 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     5701 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     5761 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     5821 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     5881 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5941 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     6001 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     6061 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     6121 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     6181 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaa
//
