LOCUS       Exported               14296 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expressed nuclear-localized GCaMP6-slow, a calcium indicator, in 
            neurons in zebrafish larva. The plasmid is in Tol2 transposon 
            backbone.
ACCESSION   .
VERSION     .
KEYWORDS    Tol2-elavl3-H2B-GCaMP6s
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14296)
  AUTHORS   Freeman J, Vladimirov N, Kawashima T, Mu Y, Sofroniew NJ, Bennett 
            DV, Rosen J, Yang CT, Looger LL, Ahrens MB
  TITLE     Mapping brain activity at scale with cluster computing.
  JOURNAL   Nat Methods. 2014 Jul 27. doi: 10.1038/nmeth.3041.
  PUBMED    25068736
REFERENCE   2  (bases 1 to 14296)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2014 Jul 27. doi: 10.1038/nmeth.3041."
FEATURES             Location/Qualifiers
     source          1..14296
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             9463..9840
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      9853..9862
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             9859..11211
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      9955..9964
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     10245..10266
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(10372..10393)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(10633..10652)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    11253..11387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11277..11296)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     11331..11350
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(12059..12075)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12059..12075)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(12112..12132)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12112..12130)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12151..12167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12151..12167)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12164..12186)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12175..12191
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(12199..12229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12244..12265
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12382..12399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12553..13141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12633..12652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13312..14172)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     13935..13954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14173..14277)
                     /gene="bla"
                     /label=AmpR promoter