LOCUS       Exported                7183 bp ds-DNA     circular SYN 02-AUG-2021
DEFINITION  Encodes GCaMP6s.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-GCaMP6s
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7183)
  TITLE     High performance microbial opsins for spatially and temporally 
            precise perturbations of large neuronal networks
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7183)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 2, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7183
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1739..3091
                     /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      1835..1844
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     2125..2146
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2252..2273)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2513..2532)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     intron          3105..3237
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3220..3239
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    3246..3834
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3299..3319)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3717..3728)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3861..3878)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3867..3978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4074..4094)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4074..4092)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4205..4224)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4270..4325
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4324..4343)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4686..4702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4686..4702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4699..4721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4710..4726
                     /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(4734..4764)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4779..4800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4859..5054
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4905..5040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4967..4986
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5060..5194
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5097..5116)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5151..5170
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5262..5279)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5433..6021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5513..5532)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6192..7052)
                     /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     6815..6834
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7053..7157)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccrgggggg gggggggggg ggggggggsg
      541 gggsggggsg rggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcggtaccat
     1741 gggttctcat catcatcatc atcatggtat ggctagcatg actggtggac agcaaatggg
     1801 tcgggatctg tacgacgatg acgataagga tctcgccacc atggtcgact catcacgtcg
     1861 taagtggaat aagacaggtc acgcagtcag agctataggt cggctgagct cactcgagaa
     1921 cgtctatatc aaggccgaca agcagaagaa cggcatcaag gcgaacttcc acatccgcca
     1981 caacatcgag gacggcggcg tgcagctcgc ctaccactac cagcagaaca cccccatcgg
     2041 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc gtgcagtcca aactttcgaa
     2101 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2161 cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatgg tgagcaaggg
     2221 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2281 ccacaagttc agcgtgtccg gcgagggtga gggcgatgcc acctacggca agctgaccct
     2341 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2401 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2461 caagtccgcc atgcccgaag gctacatcca ggagcgcacc atcttcttca aggacgacgg
     2521 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2581 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2641 cctgccggac caactgactg aagagcagat cgcagaattt aaagaggctt tctccctatt
     2701 tgacaaggac ggggatggga caataacaac caaggagctg gggacggtga tgcggtctct
     2761 ggggcagaac cccacagaag cagagctgca ggacatgatc aatgaagtag atgccgacgg
     2821 tgacggcaca atcgacttcc ctgagttcct gacaatgatg gcaagaaaaa tgaaatacag
     2881 ggacacggaa gaagaaatta gagaagcgtt cggtgtgttt gataaggatg gcaatggcta
     2941 catcagtgca gcagagcttc gccacgtgat gacaaacctt ggagagaagt taacagatga
     3001 agaggttgat gaaatgatca gggaagcaga catcgatggg gatggtcagg taaactacga
     3061 agagtttgta caaatgatga cagcgaagta actcgaggcc gcaggtaagt atcaaggtta
     3121 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc
     3181 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg
     3241 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3301 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3361 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3421 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3481 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3541 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3601 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     3661 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3721 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3781 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     3841 ttcgagctcg gtacgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3901 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3961 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     4021 ggggcaggac agcaaggggg aggattggga agacaatagc ccagcttttg ttccctttag
     4081 tgagggttaa ttgcgcgctt ggcgtaatca tkgtyatmss ykwwtycwst cctcaggtgc
     4141 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     4201 gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga
     4261 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     4321 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg
     4381 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa
     4441 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     4501 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     4561 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     4621 tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa
     4681 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4741 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4801 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc
     4861 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4921 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4981 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5041 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa
     5101 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5161 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc
     5221 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5281 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5341 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5401 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5461 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5521 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5581 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5641 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5701 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5761 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5821 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5881 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5941 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6001 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6061 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6121 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6181 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6241 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6301 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6361 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6421 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6481 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6541 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6601 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6661 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6721 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6781 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6841 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6901 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6961 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7021 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7081 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7141 caaatagggg ttccgcgcac atttccccga aaagtgccac ctg
//
