LOCUS       Exported                6150 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of hBACCS2, a human blue light-activated Ca2+ 
            channel switch.
ACCESSION   .
VERSION     .
KEYWORDS    hBACCS2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6150)
  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 6150)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6150
                     /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    2205..2793
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2258..2278)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2676..2687)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2938..3059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2975..2994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3029..3048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3066..3521)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3203..3224)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3415..3434
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3548..3652
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3654..4011
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3677..3697)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3862..3997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3924..3943
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4046..4840
                     /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(4100..4119)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4710..4729
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5028..5047)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5072..5119
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5448..6036
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5937..5956
                     /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 ctttgtcatt 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 aatcttcaga agtggctgca gctgacacat gaggtggagg
     1081 tgcaatatta caacatcaag aagcaaaatg ctgagaagca gctgctggtg gccaaggagg
     1141 gggctgagaa gataaaaaag aagagaaaca cactctttgg caccttccac gtggcccaca
     1201 gctcttccct ggatgatgta gatcataaaa ttctaacagc taagcaagca ctgagcgagg
     1261 tgacagcagc attgcgggag cgcctgcacc gctggcaaca gatcgagatc ctctgtggct
     1321 tccagattgt caacaaccct ggcatccacg gaggatccgg aggaagtgga ggaggtatcc
     1381 tgcagtctcg gggtggcagt ggtggttcag gcggttctgg atctttggct actacacttg
     1441 aacgtattga gaagaacttt gtcattactg 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 ttcagaagtg gctgcagctg acacatgagg
     1861 tggaggtgca atattacaac atcaagaagc aaaatgctga gaagcagctg ctggtggcca
     1921 aggagggggc tgagaagata aaaaagaaga gaaacacact ctttggcacc ttccacgtgg
     1981 cccacagctc ttccctggat gatgtagatc ataaaattct aacagctaag caagcactga
     2041 gcgaggtgac agcagcattg cgggagcgcc tgcaccgctg gcaacagatc gagatcctct
     2101 gtggcttcca gattgtcaac aaccctggca tccacggcgg ctctggaggt agcggactag
     2161 tttgagtcga cggtaccgcg ggcccgggat ctaagcttat cgataatcaa cctctggatt
     2221 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2281 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2341 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2401 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2461 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2521 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2581 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     2641 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     2701 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     2761 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtggatcg atccaccgga
     2821 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2881 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2941 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3001 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3061 cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3121 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3181 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3241 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3301 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3361 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3421 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3481 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3541 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3601 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3661 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3781 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3841 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3901 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3961 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4021 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4081 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4141 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4201 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4261 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4321 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4381 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4441 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4501 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4561 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4621 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4681 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4741 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4801 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4861 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4921 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4981 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     5041 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5101 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5161 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5221 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5281 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5341 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5401 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5461 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5521 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5581 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5641 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5701 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5761 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5821 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5881 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5941 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6001 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6061 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6121 tctgtggata accgtattac cgccatgcat
//
