LOCUS       Exported                5311 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  a single-wavelength GCaMP2-based genetically encoded calcium 
            indicator.
ACCESSION   .
VERSION     .
KEYWORDS    G-CaMP3
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods."; date: "2009-12"; volume: "6(12)"; pages: "875-81"
FEATURES             Location/Qualifiers
     source          1..5311
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             270..287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             291..323
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             327..350
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      357..366
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             378..434
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     647..668
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(774..795)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1035..1054)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1170..1613
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAEL
                     RHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     polyA_signal    1735..1856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1772..1791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1826..1845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1863..2318)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2000..2021)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2212..2231
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2345..2449
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2451..2808
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2474..2494)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2659..2794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2721..2740
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2843..3637
                     /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(2897..2916)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3507..3526
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     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"
     primer_bind     4734..4753
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5008..5311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"