LOCUS       40924_934        5311 bp DNA     circular SYN 13-MAY-2021
DEFINITION  a single-wavelength GCaMP2-based genetically encoded calcium 
            indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5311)
  AUTHORS   Tian L, Hires SA, Mao T, Huber D, Chiappe ME, Chalasani SH, Petreanu
            L, Akerboom J, McKinney SA, Schreiter ER, Bargmann CI, Jayaraman V, 
            Svoboda K, Looger LL.
  TITLE     Imaging neural activity in worms, flies and mice with improved GCaMP
            calcium indicators.
  JOURNAL   Nat Methods. 2009 Dec;6(12):875-81.
  PUBMED    19898485
REFERENCE   2  (bases 1 to 5311)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5311)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Methods."; date: "2009-12"; volume: "6(12)"; pages: "875-81"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             261..1610
                     /codon_start=1
                     /label=GCaMP6f
                     /note="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     polyA_signal    1735..1856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1863..2318)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2345..2449
                     /label=AmpR promoter
     promoter        2451..2808
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2843..3634
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3825..3844)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3869..3916
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4245..4833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     enhancer        5008..5311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgccaccatg atgggttctc atcatcatca tcatcatggt atggctagca
      301 tgactggtgg acagcaaatg ggtcgggatc tgtacgacga tgacgataag gatctcgcca
      361 ccatggtcga ctcatcacgt cgtaagtgga ataagacagg tcacgcagtc agagctatag
      421 gtcggctgag ctcactcgag aacgtctata tcaaggccga caagcagaag aacggcatca
      481 aggcgaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gcctaccact
      541 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
      601 gcgtgcagtc caaactttcg aaagacccca acgagaagcg cgatcacatg gtcctgctgg
      661 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtaccg
      721 gagggagcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      781 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg
      841 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      901 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      961 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacatc caggagcgca
     1021 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1081 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1141 tggggcacaa gctggagtac aacacgcgtg accaactgac tgaagagcag atcgcagaat
     1201 ttaaagaggc tttctcccta tttgacaagg acggggatgg gacaataaca accaaggagc
     1261 tggggacggt gatgcggtct ctggggcaga accccacaga agcagagctg caggacatga
     1321 tcaatgaagt agatgccgac ggtgacggca caatcgactt ccctgagttc ctgacaatga
     1381 tggcaagaaa aatgaaagac acagacagtg aagaagaaat tagagaagcg ttccgtgtgt
     1441 ttgataagga tggcaatggc tacatcagtg cagcagagct tcgccacgtg atgacaaacc
     1501 ttggagagaa gttaacagat gaagaggttg atgaaatgat cagggaagca gacatcgatg
     1561 gggatggtca ggtaaactac gaagagtttg tacaaatgat gacagcgaag tgagcggccg
     1621 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1681 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1741 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1801 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     1861 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     1921 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     1981 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2041 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2101 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2161 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2221 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2281 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     2341 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2401 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     2461 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2521 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2581 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2641 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2701 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2761 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     2821 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2881 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2941 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3001 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3061 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3121 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3181 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3241 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3301 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3361 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3421 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3481 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3541 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3601 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3661 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3721 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3781 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     3841 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     3901 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     3961 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4021 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4081 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4141 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4201 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4261 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4321 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4381 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4441 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4501 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4561 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4621 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4681 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4741 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4801 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4861 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4921 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     4981 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     5041 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     5101 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     5161 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     5221 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     5281 catctacgta ttagtcatcg ctattaccat g
//
