LOCUS       Exported                6166 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Green fluorescent reporter of Orai1-associated calcium influx.
ACCESSION   .
VERSION     .
KEYWORDS    Orai1-GCaMP6f
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6166)
  AUTHORS   Dynes JL, Amcheslavsky A, Cahalan MD
  TITLE     Genetically targeted single-channel optical recording reveals 
            multiple Orai1 gating states and oscillations in calcium influx.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jan 12;113(2):440-5. doi: 
            10.1073/pnas.1523410113. Epub 2015 Dec 28.
  PUBMED    26712003
REFERENCE   2  (bases 1 to 6166)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6166
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      922..931
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      1572..1581
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1593..1649
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1862..1883
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1989..2010)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2250..2269)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2954..3075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2991..3010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3045..3064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3082..3537)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3219..3240)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3431..3450
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3564..3668
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3670..4027
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3693..3713)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3878..4013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3940..3959
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4062..4856
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4116..4135)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4726..4745
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5044..5063)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5088..5135
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5464..6052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5953..5972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcc accatgcatc cggagcccgc cccgcccccg agccgcagca
      661 gtcccgagct tcccccaagc ggcggcagca ccaccagcgg cagccgccgg agccgccgcc
      721 gcagcgggga cggggagccc ccgggggccc cgccaccgcc gccgtccgcc gtcacctacc
      781 cggactggat cggccagagt tactccgagg tgatgagcct caacgagcac tccatgcagg
      841 cgctgtcctg gcgcaagctc tacttgagcc gcgccaagct taaagcctcc agccggacct
      901 cggctctgct ctccggcttc gccatggtgg caatggtgga ggtgcagctg gacgctgacc
      961 acgactaccc accggggctg ctcatcgcct tcagtgcctg caccacagtg ctggtggctg
     1021 tgcacctgtt tgcgctcatg atcagcacct gcatcctgcc caacatcgag gcggtgagca
     1081 acgtgcacaa tctcaactcg gtcaaggagt ccccccatga gcgcatgcac cgccacatcg
     1141 agctggcctg ggccttctcc accgtcatcg gcacgctgct cttcctagct gaggtggtgc
     1201 tgctctgctg ggtcaagttc ttgcccctca agaagcagcc aggccagcca aggcccacca
     1261 gcaagccccc cgccagtggc gcagcagcca acgtcagcac cagcggcatc accccgggcc
     1321 aggcagctgc catcgcctcg accaccatca tggtgccctt cggcctgatc tttatcgtct
     1381 tcgccgtcca cttctaccgc tcactggtta gccataagac cgaccgacag ttccaggagc
     1441 tcaacgagct ggcggagttt gcccgcttac aggaccagct ggaccacaga ggggaccacc
     1501 ccctgacgcc cggcagccac tatgcccgaa ttctgcagtc gacggtaccg cgggcccggg
     1561 atccaccggt cgccaccatg gtcgactcat cacgtcgtaa gtggaataag acaggtcacg
     1621 cagtcagagc tataggtcgg ctgagctcac tcgagaacgt ctatatcaag gccgacaagc
     1681 agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     1741 agctcgccta ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1801 acaaccacta cctgagcgtg cagtccaaac tttcgaaaga ccccaacgag aagcgcgatc
     1861 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1921 acaagggcgg taccggaggg agcatggtga gcaagggcga ggagctgttc accggggtgg
     1981 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2041 agggtgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2101 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2161 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2221 acatccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2281 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2341 aggacggcaa catcctgggg cacaagctgg agtacaacct gccggaccaa ctgactgaag
     2401 agcagatcgc agaatttaaa gaggaattct ccctatttga caaggacggg gatgggacaa
     2461 taacaaccaa ggagctgggg acggtgatgc ggtctctggg gcagaacccc acagaagcag
     2521 agctgcagga catgatcaat gaagtagatg ccgacggtga cggcacaatc gacttccctg
     2581 agttcctgac aatgatggca agaaaaatga aatacaggga cacggaagaa gaaattagag
     2641 aagcgttcgg tgtgtttgat aaggatggca atggctacat cagtgcagca gagcttcgcc
     2701 acgtgatgac aaaccttgga gagaagttaa cagatgaaga ggttgatgaa atgatcaggg
     2761 aagcagacat cgatggggat ggtcaggtaa actacgaaga gtttgtacaa atgatgacag
     2821 cgaagcttta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     2881 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2941 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3001 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3061 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     3121 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3181 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3241 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3301 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3361 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3421 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3481 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3541 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3601 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3661 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3721 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3781 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3841 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3901 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3961 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     4021 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4081 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4141 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4201 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4261 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4321 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4381 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4441 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4501 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4561 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4621 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4681 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4741 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4801 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4861 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4921 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4981 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     5041 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     5101 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     5161 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5221 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5281 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     5341 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5401 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5461 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5521 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5581 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5641 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5701 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5761 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5821 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5881 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5941 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6001 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6061 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6121 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
