LOCUS       sequence-270731-        5765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used to drive amplified GCaMP6s expression when combined with
            TET driver . Can be used to create adeno-associated virus to express
            TET-driven GCaMP6s..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5765)
  TITLE     AAV GCaMP6 Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5765)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-TRE-GCaMP6s
FEATURES             Location/Qualifiers
     source          1..5765
                     /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        173..551
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    181..199
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    217..235
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    253..271
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    289..307
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    325..343
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    361..379
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    397..415
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     460..484
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      558..567
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             573..590
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      603..612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             624..680
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     893..914
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1020..1041)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1281..1300)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1892..2480
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1945..1965)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2363..2374)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2512..2988
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2711..2730)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3028..3168
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3028..3157
                     /label=AAV2 ITR
     rep_origin      3243..3698
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3330..3349)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3540..3561
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3715..3734)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3834..3856
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3894..3912)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3980..4084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4085..4153,4154..4945)
                     /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(4303..4322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5116..5704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5605..5624
                     /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 gcgtgaggcc ctttcgtctt caagaattcc tcgagtttac
      181 tccctatcag tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta
      241 tgcagacttt actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg
      301 atagagaacg tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac
      361 tccctatcag tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta
      421 taagctttag gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca
      481 gatcgcctgg agcaattcca caacactttt gtcttatacc aactttccgt accacttcct
      541 accctcgtaa agtcgacgcc accatgggtt ctcatcatca tcatcatcat ggtatggcta
      601 gcgccaccat ggtcgactca tcacgtcgta agtggaataa gacaggtcac gcagtcagag
      661 ctataggtcg gctgagctca ctcgagaacg tctatatcaa ggccgacaag cagaagaacg
      721 gcatcaaggc gaacttccac atccgccaca acatcgagga cggcggcgtg cagctcgcct
      781 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
      841 acctgagcgt gcagtccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc
      901 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg
      961 gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1021 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg
     1081 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1141 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1201 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg
     1261 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1321 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1381 acatcctggg gcacaagctg gagtacaacc tgccggacca actgactgaa gagcagatcg
     1441 cagaatttaa agaggctttc tccctatttg acaaggacgg ggatgggaca ataacaacca
     1501 aggagctggg gacggtgatg cggtctctgg ggcagaaccc cacagaagca gagctgcagg
     1561 acatgatcaa tgaagtagat gccgacggtg acggcacaat cgacttccct gagttcctga
     1621 caatgatggc aagaaaaatg aaatacaggg acacggaaga agaaattaga gaagcgttcg
     1681 gtgtgtttga taaggatggc aatggctaca tcagtgcagc agagcttcgc cacgtgatga
     1741 caaaccttgg agagaagtta acagatgaag aggttgatga aatgatcagg gaagcagaca
     1801 tcgatgggga tggtcaggta aactacgaag agtttgtaca aatgatgaca gcgaagtagt
     1861 aaggatccaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     1921 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1981 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2041 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2101 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2161 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2221 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2281 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     2341 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2401 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2461 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     2521 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     2581 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     2641 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     2701 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     2761 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     2821 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     2881 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     2941 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     3001 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3061 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3121 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3181 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3241 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3301 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3361 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3421 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     3481 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3541 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     3601 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     3661 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     3721 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     3781 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3841 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     3901 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3961 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4021 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4081 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4141 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4201 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4261 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     4321 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     4381 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     4441 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     4501 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     4561 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     4621 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     4681 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     4741 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4801 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4861 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4921 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4981 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5041 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5101 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5161 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5221 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5281 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5341 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5401 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5461 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5521 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5581 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5641 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5701 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5761 catgt
//
