LOCUS       sequence-207203-        6461 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 6461)
  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 6461)
  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..6461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        406..683
                     /label=chicken beta-actin promoter
     primer_bind     963..985
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1047..1066
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1165..1174
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1171..1179,1180..1197,1198..1200,1201..1233,1234..1236,
                     1237..1260,1261..1287,1288..1344,1345..1350,1351..1620,
                     1621..1638,1639..2073,2074..2079,2080..2523)
                     /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      1267..1276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1557..1578
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1684..1705)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1945..1964)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2588..3176
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2641..2661)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3059..3070)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3208..3684
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3407..3426)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3724..3864
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3724..3853
                     /label=AAV2 ITR
     rep_origin      3939..4394
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4026..4045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4236..4257
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4411..4430)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4530..4552
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4590..4608)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4676..4780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4781..4849,4850..5641)
                     /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(4999..5018)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5812..6400
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6301..6320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtccatt gacgtcaata atgacgtatg
      181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      361 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
      421 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
      481 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg
      541 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
      601 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      661 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgacgct gccttcgccc cgtgccccgc
      721 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      781 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
      841 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
      901 gggagcggct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc
      961 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     1021 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     1081 tggcaaagaa ttggtaccgg atcctctaga gtcgactccg gaacaacttc gtatatacct
     1141 ttctatacga agttgtggcg cgccgccacc atgggttctc atcatcatca tcatcatggt
     1201 atggctagca tgactggtgg acagcaaatg ggtcgggatc tgtacgacga tgacgataag
     1261 gatctcgcca ccatggtcga ctcatcacgt cgtaagtgga ataagacagg tcacgcagtc
     1321 agagctatag gtcggctgag ctcactcgag aacgtctata tcaaggccga caagcagaag
     1381 aacggcatca aggcgaactt ccacatccgc cacaacatcg aggacggcgg cgtgcagctc
     1441 gcctaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1501 cactacctga gcgtgcagtc caaactttcg aaagacccca acgagaagcg cgatcacatg
     1561 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1621 ggcggtaccg gagggagcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1681 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt
     1741 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1801 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1861 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacatc
     1921 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1981 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2041 ggcaacatcc tggggcacaa gctggagtac aacctgccgg accaactgac tgaagagcag
     2101 atcgcagaat ttaaagaggc tttctcccta tttgacaagg acggggatgg gacaataaca
     2161 accaaggagc tggggacggt gatgcggtct ctggggcaga accccacaga agcagagctg
     2221 caggacatga tcaatgaagt agatgccgac ggtgacggca caatcgactt ccctgagttc
     2281 ctgacaatga tggcaagaaa aatgaaatac agggacacgg aagaagaaat tagagaagcg
     2341 ttcggtgtgt ttgataagga tggcaatggc tacatcagtg cagcagagct tcgccacgtg
     2401 atgacaaacc ttggagagaa gttaacagat gaagaggttg atgaaatgat cagggaagca
     2461 gacatcgatg gggatggtca ggtaaactac gaagagtttg tacaaatgat gacagcgaag
     2521 tgagctagca caacttcgta tatacctttc tatacgaagt tgtgaattcg atatcaagct
     2581 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2641 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2701 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2761 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2821 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     2881 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2941 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3001 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3061 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3121 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3181 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     3241 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     3301 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     3361 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     3421 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     3481 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     3541 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     3601 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3661 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
     3721 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3781 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3841 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     3901 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     3961 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     4021 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     4081 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     4141 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4201 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4261 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     4321 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     4381 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     4441 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     4501 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     4561 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     4621 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     4681 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4741 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4801 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4861 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4921 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4981 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5041 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5101 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5161 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5221 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5281 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5341 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5401 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5461 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5521 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5581 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5641 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5701 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5761 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5821 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5881 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5941 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6001 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6061 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6121 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6181 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6241 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6301 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6361 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6421 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
