LOCUS       Exported                7807 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of dmBACCS2VL, a modified Drosophila 
            melanogaster blue light-activated Ca2+ channel switch, and dOrai.
ACCESSION   .
VERSION     .
KEYWORDS    dmBACCS2VL-IRES-dOrai
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7807)
  AUTHORS   Ishii T, Sato K, Kakumoto T, Miura S, Touhara K, Takeuchi S, Nakata 
            T
  TITLE     Light generation of intracellular Ca(2+) signals by a genetically 
            encoded protein BACCS.
  JOURNAL   Nat Commun. 2015 Aug 18;6:8021. doi: 10.1038/ncomms9021.
  PUBMED    26282514
REFERENCE   2  (bases 1 to 7807)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7807
                     /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      603..612
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    2186..2759
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2209..2761
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2353..2370)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2580..2599
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      3281..3290
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    3862..4450
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3915..3935)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4333..4344)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4595..4716
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4632..4651)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4686..4705
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4723..5178)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4860..4881)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5072..5091
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5205..5309
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5311..5668
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(5334..5354)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5519..5654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5581..5600
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5703..6497
                     /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(5757..5776)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6367..6386
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6685..6704)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6729..6776
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7105..7693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7594..7613
                     /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 gctagcctcg
      601 aggccaccat gggaccggtc ggaggaagcg gtggatcttt ggctactaca cttgaacgta
      661 ttgagaagaa ctttctcatt actgacccaa gattgccaga taatcccatt atattcgcgt
      721 ccgatagttt cttgcagttg acagaatata gccgtgaaga aattttggga agaaactgca
      781 ggtttctaca aggtcctgaa actgatcgcg cgacagtgag aaaaattaga gatgccatag
      841 ataaccaaac agaggtcact gttcagctga ttaattatac aaagagtggt aaaaagttct
      901 ggaacctctt tcacttgcag cctatgcgag atcagaaggg agatgtccag tactttattg
      961 gggttcagtt ggatggaact gagcatgtcc gagatgctgc cgagagagag ggagtcatgc
     1021 tgattaagaa aactgcagaa aatctgcaat catggcttca atacacatat gaactagaaa
     1081 gtaagaatca tcagaagaag cgcacgtcgg ctgagaagca gctacagtcg gccagagagg
     1141 cttgtgagaa attgcgtaag aaacggtcaa gtttggtggg tgcgttcgtt tccacgcacg
     1201 gaaagagtat tgatgatgtg gatcggtcga ttgttgaggc acggaatgcc ctcggagatg
     1261 taacaaacga gctgcaagaa cgactgcatc gctggaagca aatcgagacg tgccttggct
     1321 taaacattgt gaacaacaat ggtctgcccg gaggatccgg aggaagtgga ggaggtatcc
     1381 tgcagtctcg gggtggcagt ggtggttcag gcggttctgg atctttggct actacacttg
     1441 aacgtattga gaagaacttt ctcattactg acccaagatt gccagataat cccattatat
     1501 tcgcgtccga tagtttcttg cagttgacag aatatagccg tgaagaaatt ttgggaagaa
     1561 actgcaggtt tctacaaggt cctgaaactg atcgcgcgac agtgagaaaa attagagatg
     1621 ccatagataa ccaaacagag gtcactgttc agctgattaa ttatacaaag agtggtaaaa
     1681 agttctggaa cctctttcac ttgcagccta tgcgagatca gaagggagat gtccagtact
     1741 ttattggggt tcagttggat ggaactgagc atgtccgaga tgctgccgag agagagggag
     1801 tcatgctgat taagaaaact gcagaaaatc tgcaatcatg gcttcaatac acatatgaac
     1861 tagaaagtaa gaatcatcag aagaagcgca cgtcggctga gaagcagcta cagtcggcca
     1921 gagaggcttg tgagaaattg cgtaagaaac ggtcaagttt ggtgggtgcg ttcgtttcca
     1981 cgcacggaaa gagtattgat gatgtggatc ggtcgattgt tgaggcacgg aatgccctcg
     2041 gagatgtaac aaacgagctg caagaacgac tgcatcgctg gaagcaaatc gagacgtgcc
     2101 ttggcttaaa cattgtgaac aacaatggtc tgcccggcgg ctctggaggt agcggactag
     2161 tttgagtcga gccgcggcaa ttccgcccct ctccctcccc cccccctaac gttactggcc
     2221 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     2281 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     2341 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     2401 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     2461 accccccacc tggcaacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     2521 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     2581 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     2641 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     2701 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     2761 atgtctgtgt ggaccacggc caacaattcg ggcctagaga cgccaacaaa gtcgccgatc
     2821 acgtcgtcgg ttccgcgggc atcgagaagt tctgcggtga tcaccactgg aaatcaccag
     2881 cagcaccact tccagcacgt tgtggccgcc gccgttgcag ccgccacttc agtggccacc
     2941 ggtcatcagt tccagcagca gttcccgctg cacgcacatc cgcatccgca tccacagcac
     3001 cacagcaaca gtcccacggg tagcggcagc aacagcaaca acagcgcagg tttccagcgc
     3061 accagcatca gcaactcgct gctgcagttc ccgccgcccc cgcccccttc ctcacagaac
     3121 caggcgaagc cgcgcggaca ccatcgcacc gccagcagca gcatgtcgca atccggcgag
     3181 gatctccact cgcccaccta cctgtcctgg cggaagctcc agcttagtcg ggccaagcta
     3241 aaggcatcca gcaagacgtc cgccctgctc tccggattcg ccatggtggc gatggtggag
     3301 gtgcaactgg atcacgatac gaatgtacca ccgggcatgc tgatagcctt cgccatctgc
     3361 accaccctcc tggtggcggt gcacatgctg gccctgatga tcagcacctg tatcctgcca
     3421 aatattgaga cggtgtgtaa tttgcatagt atttccctgg tccacgaatc gccacacgag
     3481 cgtctccatt ggtacattga aacggcgtgg gcattctcca cgctgctggg actcatactc
     3541 tttctactgg agatagccat cctgtgctgg gtgaaattct atgacctaag tccgccggcg
     3601 gcttggtcag cgtgtgtggt cctcataccg gtgatgatca tctttatggc cttcgccatt
     3661 cacttctatc gctccctggt gtcgcacaaa tatgaggtga cagtctcggg catccgtgag
     3721 ctggagatgc tcaaggagca gatggagcag gatcatttgg agcatcacaa taacatacga
     3781 aataatggca tgaactatgg cgcatccggg gacattgtct aggtcgacgg taccgcgggc
     3841 ccgggatcta agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3901 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3961 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4021 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4081 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4141 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     4201 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     4261 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     4321 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     4381 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4441 gcctccccgc atcgataccg tggatcgatc caccggatct agataactga tcataatcag
     4501 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     4561 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     4621 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4681 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     4741 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     4801 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4861 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4921 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4981 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     5041 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     5101 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     5161 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5221 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5281 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     5341 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5401 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5461 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5521 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5581 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     5641 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     5701 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5761 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5821 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5881 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5941 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6001 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6061 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6121 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6181 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     6241 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6301 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6361 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6421 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6481 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6541 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     6601 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     6661 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     6721 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     6781 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     6841 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     6901 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     6961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     7261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7741 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7801 catgcat
//
