LOCUS       sequence-212310-        6450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-off expression of GCaMP6s.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6450)
  AUTHORS   Markowitz JE, Gillis WF, Beron CC, Neufeld SQ, Robertson K, Bhagat 
            ND, Peterson RE, Peterson E, Hyun M, Linderman SW, Sabatini BL, 
            Datta SR
  TITLE     The Striatum Organizes 3D Behavior via Moment-to-Moment Action 
            Selection.
  JOURNAL   Cell. 2018 May 10. pii: S0092-8674(18)30512-9. doi: 
            10.1016/j.cell.2018.04.019.
  PUBMED    29779950
REFERENCE   2  (bases 1 to 6450)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2018 
            May 10. pii: S0092-8674(18)30512-9. doi: 
            10.1016/j.cell.2018.04.019."
COMMENT     Sequence Label: pAAV-CBA-DO(FAS)-GCaMP6s
FEATURES             Location/Qualifiers
     source          1..6450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        395..672
                     /label=chicken beta-actin promoter
     primer_bind     952..974
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1036..1055
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1154..1163
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1160..1168,1169..1186,1187..1189,1190..1222,1223..1225,
                     1226..1249,1250..1276,1277..1333,1334..1339,1340..1609,
                     1610..1627,1628..2062,2063..2068,2069..2512)
                     /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      1256..1265
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1546..1567
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1673..1694)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1934..1953)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2577..3165
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2630..2650)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3048..3059)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3197..3673
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3396..3415)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3713..3853
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3713..3842
                     /label=AAV2 ITR
     rep_origin      3928..4383
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4015..4034)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4225..4246
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4400..4419)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4519..4541
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4579..4597)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4665..4769
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4770..4838,4839..5630)
                     /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     complement(4988..5007)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5801..6389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6290..6309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
      421 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
      481 ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg
      541 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
      601 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      661 gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc gtgccccgct ccgccgccgc
      721 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      781 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
      841 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
      901 ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct
      961 tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt
     1021 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     1081 tggtaccgga tcctctagag tcgactccgg aacaacttcg tatatacctt tctatacgaa
     1141 gttgtggcgc gccgccacca tgggttctca tcatcatcat catcatggta tggctagcat
     1201 gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg atctcgccac
     1261 catggtcgac tcatcacgtc gtaagtggaa taagacaggt cacgcagtca gagctatagg
     1321 tcggctgagc tcactcgaga acgtctatat caaggccgac aagcagaaga acggcatcaa
     1381 ggcgaacttc cacatccgcc acaacatcga ggacggcggc gtgcagctcg cctaccacta
     1441 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1501 cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1561 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg
     1621 agggagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     1681 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc
     1741 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1801 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     1861 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac
     1921 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1981 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2041 ggggcacaag ctggagtaca acctgccgga ccaactgact gaagagcaga tcgcagaatt
     2101 taaagaggct ttctccctat ttgacaagga cggggatggg acaataacaa ccaaggagct
     2161 ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc aggacatgat
     2221 caatgaagta gatgccgacg gtgacggcac aatcgacttc cctgagttcc tgacaatgat
     2281 ggcaagaaaa atgaaataca gggacacgga agaagaaatt agagaagcgt tcggtgtgtt
     2341 tgataaggat ggcaatggct acatcagtgc agcagagctt cgccacgtga tgacaaacct
     2401 tggagagaag ttaacagatg aagaggttga tgaaatgatc agggaagcag acatcgatgg
     2461 ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagt gagctagcac
     2521 aacttcgtat atacctttct atacgaagtt gtgaattcga tatcaagctt atcgataatc
     2581 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2641 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2701 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2761 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2821 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2881 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2941 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     3001 atgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3061 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3121 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc
     3181 tgctcgagag atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     3241 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     3301 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     3361 gcaagttggg aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc
     3421 agtggcacaa tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc
     3481 tcagcctccc gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt
     3541 tttttggtag agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca
     3601 ggtgatctac ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc
     3661 cttccctgtc cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc
     3721 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     3781 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     3841 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3901 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     3961 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     4021 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4081 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4141 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4201 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     4261 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     4321 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     4381 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     4441 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4501 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4561 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     4621 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4681 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4741 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     4801 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4861 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     4921 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     4981 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5041 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5101 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5161 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5221 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5281 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     5341 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5401 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5461 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5521 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5581 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5641 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5701 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5761 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5821 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5881 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5941 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6001 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6061 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6121 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6181 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6241 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6301 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6361 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6421 tggccttttg ctggcctttt gctcacatgt
//
