LOCUS       sequence-485696-        6724 bp    DNA     circular SYN 10-SEP-2025
DEFINITION  Cation channelrhodopsin ChroME2f fused to GCaMP8m and targeted to 
            the neuronal soma and proximal dendrites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6724)
  AUTHORS   Adesnik H
  TITLE     Miscellaneous Optogenetic Tools
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6724)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV_hSyn_ChroME2f_GCaMP8m_ST
FEATURES             Location/Qualifiers
     source          1..6724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   826..955
                     /label=AAV2 ITR
     polyA_signal    1117..1238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1154..1173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1208..1227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1267..1283
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1268..1284
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1287..1875
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1393..1404
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1802..1822
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     2654..2673
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2913..2934
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3040..3061)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3343..3360)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(4378..4825)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   4907..5036
                     /label=AAV2 ITR
     rep_origin      5108..5563
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5195..5214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5405..5426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5589..5693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5694..5762,5763..6554)
                     /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(5912..5931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg cgctcgctcg
      841 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
      901 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
      961 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatccttatc
     1021 gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca tctccccctg
     1081 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1201 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc gaagcggccg
     1261 cccgggtcga ggtcgacggt atcgatgcgg ggaggcggcc caaagggaga tccgactcgt
     1321 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg
     1381 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca
     1441 ggtggcaaca caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga
     1501 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     1561 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt
     1621 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg
     1681 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga
     1741 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     1801 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     1861 atccagaggt tgattatcga taagcttgat atcttagaat ctggtggctt cagggaagtc
     1921 cgtggcacag ctgatgaagc tgtcaatgct ggacatgctc cgcatgtcga tgacgccctc
     1981 cgtgcgtgcg ggcagagggg ccacggggct gggcatgctg ctcatctcca gctcctctgg
     2041 aggcttgctc tgcggggcca tctccttggt gttgaggatg ggctgggact gcaattgctt
     2101 cgctgtcatc atttgtacaa actcttcgta gtttacctga ccatccccat cgatgtctgc
     2161 ttccctgatc atttcatcaa cctcttcatc tgttaacttc tctccaaggt ttgtcatcac
     2221 gtggcgaagc tctgctgcac tgatgtagcc attgccatcc ttatcaaaca caccgaacgc
     2281 ttctctaatt tcttcttccg tgtccctgta tttcattttt cttgccatca ttgtcaggaa
     2341 ctcagggaag tcgattgtgc cgtcaccgtc ggcatctact tcattgatca tgtcctgcag
     2401 ctctgcttct gtggggttct gccccagaga ccgcatcacc gtccccagct ccttggttgt
     2461 tattgtccca tccccgtcct tgtcaaatag ggagaaagcc tctttgtatt ctgcgatctg
     2521 ctcttcagtc agttggtccg gcaggttgta ctccagcttg tgccccagga tgttgccgtc
     2581 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
     2641 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggat
     2701 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
     2761 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
     2821 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcacc
     2881 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     2941 caccaccccg gtgaacagct cctcgccctt gctcaccatg ctccctccgg taccgccctt
     3001 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
     3061 gtgatcgcgc ttctcgttgg ggtctttcga aagtttggac tccacgctca ggtagtggtt
     3121 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtaggcgag
     3181 ctgcacgccg ccgtcctcga tgttgtggcg gatgtggaag ttcgccttga tgccgttctt
     3241 ctgcttgtcg gccttgatat agacgttgat cttcaggccc atcaggcggc cgatgatctt
     3301 cacggcggtg gccacctcct tgaaggtctt cttgcgacgc gtgtgatgat ggtgatgatg
     3361 ggcggccgcc actcctccct cctcctcctc gtcgacgaag gtctcgatct ccatgttctc
     3421 gccggccacg ttgatggtgg ttgtcttccg gatgtcgccg tggatcagga tgtgctcgtg
     3481 gatcttcact ctcaggtggt ggcccaggaa gccccaagtg ttcttggcga tcagatccag
     3541 gatcaggtgg ccgattccgc tggtgtaagg tgtgatcagg cccagtccct cgtgtccggc
     3601 gatgaacatc acggggaaca tcagccaaga gcagaagaag acgtaggcca tgatcttgca
     3661 aatcttccgg cagacgcctt tgggcagggt gtggtagctc tcgatataca ccttggcgat
     3721 ctggaagaag caagtcacgc cgtagaacag gccgatcatg aagaacagga tcttcagggg
     3781 gccctttgta aaggcggctg tgatccccca cacgatgttg ccgatgtcgg tcaccaggat
     3841 ggtcatggtc cgcttgctgt actcttcgtg cagtccggtc aggttgctca ggtggatcag
     3901 gatcacgggg caagtcagga gccactcgct gtaccgcagc caagccacgg ctcctccgtt
     3961 ggtctggtac actgtggctg ggctgtcgac atcgtggaac agctcgatga agcacttgac
     4021 gagctcgatc acgcagacgt agacctcctc ccagccggtt gtggccttga agctgtgcca
     4081 accgtagaag atgagctgca cgatagccac gatcacggtg aaccactgga gtcccactgc
     4141 gatcttgtgc tgcagctcgg ttccgtggtt gatgtggggg aagcaaccgt gatcggctcc
     4201 ggctgttgtg gcgttagagg tttctccgtg agcgtctgct gctggtgtca ccactgctgc
     4261 ggcgctcagc aggcctctaa ttgtggcttc ggcgctaggc acggctgaga taaaggcgtg
     4321 ggtcattgtg gcggctgttt ccatactagc ggatccttgc tatcagcttg aattcgactg
     4381 cgctctcagg cacgacacga ctcctccgct gcccaccgca gactgaggca gcgctgagtc
     4441 gccggcgccg cagcgcagat ggtcgcgccc gtgcccccct atctcgcgcc tcgcgtggtg
     4501 cggtccggct gggccggcgg cggcgcggac gcgaccaagg tggccgggaa ggggagtttg
     4561 cgggggaccg gcgagtgacg tcagcgcgcc ttcagtgctg aggcggcggt ggcgcgcgcc
     4621 gccaggcggg ggcgaaggca ctgtccgcgg tgctgaagct ggcagtgcgc acgcgcctcg
     4681 ccgcatcctg tttcccctcc ccctctctga taggggatgc gcaatttggg gaatgggggt
     4741 tgggtgcttg tccagtgggt cggggtcggt cgtcaggtag gcacccccac cccgcctcat
     4801 cctggtccta aaacccactt gcactcatac gcagggccct ctgcagagat cttcctagag
     4861 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     4921 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     4981 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgg
     5041 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     5101 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5161 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5221 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5281 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5341 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5401 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5461 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5521 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     5581 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5641 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5701 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5761 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5821 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5881 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5941 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6001 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6061 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6121 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6181 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6241 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6301 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6361 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6421 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6481 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6541 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6601 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6661 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6721 cttc
//
