LOCUS       sequence-190132-        7016 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express Orai1 in  mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7016)
  AUTHORS   Sun J, Lu F, He H, Shen J, Messina J, Mathew R, Wang D, Sarnaik AA, 
            Chang WC, Kim M, Cheng H, Yang S
  TITLE     STIM1- and Orai1-mediated Ca(2+) oscillation orchestrates 
            invadopodium formation and melanoma invasion.
  JOURNAL   J Cell Biol. 2014 Nov 24;207(4):535-48. doi: 10.1083/jcb.201407082. 
            Epub 2014 Nov 17.
  PUBMED    25404747
REFERENCE   2  (bases 1 to 7016)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1083/jcb.201407082"; journalName: "J Cell Biol"; date: 
            "2014-11-24- 24"; volume: "207"; issue: "4"; pages: "535-48"
COMMENT     Sequence Label: pLNCX2-Orai1
FEATURES             Location/Qualifiers
     source          1..7016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      853..862
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             1542..2135
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2238..2260)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2346..2486
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2401..2420
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2501..2518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2672..3260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2752..2771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3431..4222,4223..4291))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4054..4073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4292..4396)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4464..4482
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5054..5229
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5292..5491
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5692..6108
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6038..6060
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6076..6092
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             6152..6946
                     /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(6206..6225)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6816..6835
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atccatgcat ccggagcctg ccccgccccc gagccgcagc agtcccgagc
      601 ttcccccaag cggcggcagc accaccagcg gcagccgccg gagccgccgc cgcagcgggg
      661 acggggagcc cccgggggcc ccgccaccgc cgccgtccgc cgtcacctac ccggactgga
      721 tcggccagag ttactccgag gtgatgagcc tcaacgagca ctccatgcag gcgctgtcct
      781 ggcgcaagct ctacttgagc cgcgccaagc ttaaagcctc cagccggacc tcggctctgc
      841 tctccggctt cgccatggtg gcaatggtgg aggtgcagct ggacgctgac cacgactacc
      901 caccggggct gctcatcgcc ttcagtgcct gcaccacagt gctggtggct gtgcacctgt
      961 ttgcgctcat gatcagcacc tgcatcctgc ccaacatcga ggcggtgagc aacgtgcata
     1021 acctgaattc ggtcaaggag tccccccatg agcgcatgca ccgccacatc gagctggcct
     1081 gggccttctc caccgtcatc ggcacgctgc tcttcctagc tgaggtggtg ctgctctgct
     1141 gggtcaagtt cttgcccctc aagaagcagc caggccagcc aaggcccacc agcaagcccc
     1201 ccgccagtgg cgcagcagcc aacgtcagca ccagcggcat caccccgggc caggcagctg
     1261 ccatcgcctc gaccaccatc atggtgccct tcggcctgat ctttatcgtc ttcgccgtcc
     1321 acttctaccg ctcactggtt agccataaga ccgaccgaca gttccaggag ctcaacgatc
     1381 tggcggagtt tgcccgctta caggaccagc tggaccacag aggggaccac cccctgacgc
     1441 ccggcagcca ctatgccctc gagcggccgc ttgtcgacag gccttaatgg cctaacatcg
     1501 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     1561 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     1621 agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
     1681 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
     1741 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     1801 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
     1861 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     1921 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     1981 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
     2041 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     2101 agtgattgac tacccgtcag cgggggtctt tcatttgggg gctcgtccgg gatcgggaga
     2161 cccctgccca gggaccaccg acccaccacc gggaggtaag ctggctgcct cgcgcgtttc
     2221 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     2281 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     2341 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     2401 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     2461 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
     2521 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2581 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2641 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2701 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2761 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2821 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2881 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2941 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3001 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3061 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3121 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3181 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3241 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3301 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3361 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3421 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3481 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3541 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3601 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3661 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3721 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
     3781 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3841 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3901 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3961 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4021 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     4081 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4141 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4201 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4261 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4321 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4381 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4441 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat
     4501 tcataccaga tcaccgaaaa ctgtcctcca aatgtgtccc cctcacactc ccaaattcgc
     4561 gggcttctgc ctcttagacc actctaccct attccccaca ctcaccggag ccaaagccgc
     4621 ggcccttccg tttctttgct tttgaaagac cccacccgta ggtggcaagc tagcttaagt
     4681 aacgccactt tgcaaggcat ggaaaaatac ataactgaga atagaaaagt tcagatcaag
     4741 gtcaggaaca aagaaacagc tgaataccaa acaggatatc tgtggtaagc ggttcctgcc
     4801 ccggctcagg gccaagaaca gatgagacag ctgagtgatg ggccaaacag gatatctgtg
     4861 gtaagcagtt cctgccccgg ctcggggcca agaacagatg gtccccagat gcggtccagc
     4921 cctcagcagt ttctagtgaa tcatcagatg tttccagggt gccccaagga cctgaaaatg
     4981 accctgtacc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     5041 cgctctccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcttccgat
     5101 agactgcgtc gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat
     5161 cgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc acgacggggg
     5221 tctttcattt gggggctcgt ccgggatttg gagacccctg cccagggacc accgacccac
     5281 caccgggagg taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg
     5341 tttgatgtta tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc
     5401 cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac
     5461 tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg
     5521 tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
     5581 tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc
     5641 gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact
     5701 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
     5761 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
     5821 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
     5881 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
     5941 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
     6001 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
     6061 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg aattccgatc
     6121 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     6181 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     6241 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     6301 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     6361 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     6421 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     6481 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     6541 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     6601 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     6661 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6721 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     6781 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6841 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6901 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     6961 ttcgaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccg
//
