LOCUS       sequence-255909-        7902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bicistronic AAV vector to express GCaMP6m and soma-targeted ChRmine 
            under the control of CamKIIa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7902)
  AUTHORS   Marshel JH, Kim YS, Machado TA, Quirin S, Benson B, Kadmon J, Raja 
            C, Chibukhchyan A, Ramakrishnan C, Inoue M, Shane JC, McKnight DJ, 
            Yoshizawa S, Kato HE, Ganguli S, Deisseroth K
  TITLE     Cortical layer-specific critical dynamics triggering perception.
  JOURNAL   Science. 2019 Aug 9;365(6453). pii: science.aaw5202. doi: 
            10.1126/science.aaw5202. Epub 2019 Jul 18.
  PUBMED    31320556
REFERENCE   2  (bases 1 to 7902)
  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: "Science."; 
            date: "2019-08-9"; volume: "365(6453). pii: science.aaw5202. doi"; 
            pages: " 10.1126/science.aaw5202. Epub 2019 Jul 18"
COMMENT     Sequence Label: pAAV-CaMKIIa-GCaMP 6m-p2A-ChRmine-Kv2.1-WPRE
FEATURES             Location/Qualifiers
     source          1..7902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        154..1442
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1463..1472
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1484..1540
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1753..1774
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1880..1901)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2141..2160)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2732..2788
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3980..4006
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    4034..4622
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4087..4107)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4505..4516)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4654..5130
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4853..4872)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5170..5310
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5170..5299
                     /label=AAV2 ITR
     rep_origin      5385..5840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5472..5491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5682..5703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5857..5876)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5976..5998
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6036..6054)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6122..6226
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6227..6295,6296..7087)
                     /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(6445..6464)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7258..7846
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7747..7766
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgtt taacattatg gccttaggtc acttcatctc
      181 catggggttc ttcttctgat tttctagaaa atgagatggg ggtgcagaga gcttcctcag
      241 tgacctgccc agggtcacat cagaaatgtc agagctagaa cttgaactca gattactaat
      301 cttaaattcc atgccttggg ggcatgcaag tacgatatac agaaggagtg aactcattag
      361 ggcagatgac caatgagttt aggaaagaag agtccagggc agggtacatc tacaccaccc
      421 gcccagccct gggtgagtcc agccacgttc acctcattat agttgcctct ctccagtcct
      481 accttgacgg gaagcacaag cagaaactgg gacaggagcc ccaggagacc aaatcttcat
      541 ggtccctctg ggaggatggg tggggagagc tgtggcagag gcctcaggag gggccctgct
      601 gctcagtggt gacagatagg ggtgagaaag cagacagagt cattccgtca gcattctggg
      661 tctgtttggt acttcttctc acgctaaggt ggcggtgtga tatgcacaat ggctaaaaag
      721 cagggagagc tggaaagaaa caaggacaga gacagaggcc aagtcaacca gaccaattcc
      781 cagaggaagc aaagaaacca ttacagagac tacaaggggg aagggaagga gagatgaatt
      841 agcttcccct gtaaacctta gaacccagct gttgccaggg caacggggca atacctgtct
      901 cttcagagga gatgaagttg ccagggtaac tacatcctgt ctttctcaag gaccatccca
      961 gaatgtggca cccactagcc gttaccatag caactgcctc tttgccccac ttaatcccat
     1021 cccgtctgtt aaaagggccc tatagttgga ggtgggggag gtaggaagag cgatgatcac
     1081 ttgtggacta agtttgttcg catccccttc tccaaccccc tcagtacatc accctggggg
     1141 aacagggtcc acttgctcct gggcccacac agtcctgcag tattgtgtat ataaggccag
     1201 ggcaaagagg agcaggtttt aaagtgaaag gcaggcaggt gttggggagg cagttaccgg
     1261 ggcaacggga acagggcgtt tcggaggtgg ttgccatggg gacctggatg ctgacgaagg
     1321 ctcgcgaggc tgtgagcagc cacagtgccc tgctcagaag ccccaagctc gtcagtcaag
     1381 ccggttctcc gtttgcactc aggagcacgg gcaggcgagt ggcccctagt tctgggggca
     1441 gctctagagc ggtaccggat ccgccaccat ggtggactca tcacgtcgta agtggaataa
     1501 gacaggtcac gcagtcagag ctataggtcg gctgagctca ctcgagaacg tctatatcaa
     1561 ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga
     1621 cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt
     1681 gctgctgccc gacaaccact acctgagcgt gcagtccaaa ctttcgaaag accccaacga
     1741 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1801 ggacgagctg tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt
     1861 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1921 cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1981 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     2041 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2101 gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2161 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2221 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacc tgccggacca
     2281 actgactgaa gagcagatcg cagaatttaa agaggctttc tccctatttg acaaggacgg
     2341 ggatgggaca ataacaacca aggagctggg gacggtgatg cggtctctgg ggcagaaccc
     2401 cacagaagca gagctgcagg acatgatcaa tgaagtagat gccgacggtg acggcacaat
     2461 cgacttccct gagttcctga caatgatggc aagaaaaggg agctacaggg acacggaaga
     2521 agaaattaga gaagcgttcg gtgtgtttga taaggatggc aatggctaca tcagtgcagc
     2581 agagcttcgc cacgtgatga caaaccttgg agagaagtta acagatgaag aggttgatga
     2641 aatgatcagg gaagcagaca tcgatgggga tggtcaggta aactacgaag agtttgtaca
     2701 aatgatgaca gcgaagacta gtggctccgg agccacgaac ttctctctgt taaagcaagc
     2761 aggagacgtg gaagaaaacc ccggtcccat ggcacacgca ccaggcaccg accagatgtt
     2821 ctacgtgggc acaatggacg gctggtatct ggataccaag ctgaactccg tggccatcgg
     2881 cgcccactgg tcttgcttta tcgtgctgac aatcaccaca ttctacctgg gctatgagtc
     2941 ctggacatct cggggcccta gcaagagaac ctccttttac gccggctatc aggaggagca
     3001 gaacctggcc ctgtttgtga atttctttgc catgctgagc tacttcggca agatcgtggc
     3061 cgacacactg ggccacaact tcggcgatgt gggccccttc atcatcggct tcggcaatta
     3121 caggtatgcc gactacatgc tgacctgccc aatgctggtg tacgatctgc tgtatcagct
     3181 gagggccccc tatcgcgtgt cttgtagcgc catcatcttt gccatcctga tgtctggcgt
     3241 gctggccgag ttctacgcag agggcgaccc taggctgagg aatggcgcct acgcctggta
     3301 tggctttggc tgtttctggt ttatcttcgc ctactctatc gtgatgagca tcgtggccaa
     3361 gcagtatagc cggctggccc agctggccca ggatacaggc gccgagcaca gcctgcacgt
     3421 gctgaagttc gccgtgttta ccttctccat gctgtggatt ctgtttcccc tggtgtgggc
     3481 catctgccct agaggcttcg gctggatcga cgataactgg acagaggtgg cccactgcgt
     3541 gtgcgacatc gtggccaagt cctgttacgg ctttgccctg gcccggttca gaaagaccta
     3601 tgatgaggag ctgtttcggc tgctggagca gctgggacac gacgaggatg agttccagaa
     3661 gctggagctg gatatgaggc tgagcagcaa tggcgagcgc ctgcggagac tgtctgcggc
     3721 cgccaagagc aggatcacca gcgagggcga gtacatcccc ctggaccaga tcgacatcaa
     3781 cgtgcagtcc cagcccatcc tcaacaccaa ggagatggcc ccgcagagca agcctccaga
     3841 ggagctggag atgagcagca tgcccagccc cgtggcccct ctgcccgcac gcacggaggg
     3901 cgtcatcgac atgcggagca tgtccagcat tgacagcttc atcagctgtg ccacggactt
     3961 ccctgaagcc accagattct acccatacga tgttccagat tacgcttaag aattcgatat
     4021 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4081 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4141 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4201 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4261 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg gaactttcgc
     4321 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4381 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     4441 tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4501 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4561 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4621 gcatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca
     4681 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa
     4741 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg
     4801 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg
     4861 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt
     4921 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg
     4981 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc
     5041 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag
     5101 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg
     5161 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     5221 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     5281 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
     5341 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
     5401 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5461 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5521 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5581 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5641 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5701 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     5761 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5821 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     5881 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5941 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     6001 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     6061 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6121 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6181 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6241 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6301 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6361 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6421 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6481 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6541 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6601 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6661 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6721 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6781 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6841 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6901 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6961 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7021 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7081 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7141 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7201 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7261 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7321 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7381 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     7441 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7501 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7561 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7621 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7681 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7741 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7801 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7861 ggccttttta cggttcctgg ccttttgctg gccttttgct ca
//
