LOCUS       sequence-457770-        7066 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Cation channelrhodopsin ChRmine targeted to the neuronal soma and 
            proximal dendrites in a viral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7066)
  AUTHORS   Adesnik H
  TITLE     Miscellaneous Optogenetic Tools
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7066)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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            "Unpublished"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-hSyn-DIO-GCaMP6m-p2A-ChRmine-ST
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1162..1617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1511..1530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1878..2325
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(2371..2404)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2455..2488)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2500..2526)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3718..3774)
                     /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"
     primer_bind     4346..4365
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4605..4626
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4732..4753)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(4966..5022)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     regulatory      5034..5043
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    5050..5083
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    5134..5167
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5192..5780
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5245..5265)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5663..5674)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5783..5799)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5784..5800)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5829..5950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5840..5859)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5894..5913
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6112..6241
                     /label=AAV2 ITR
     rep_origin      complement(6478..7066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6558..6577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     1621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1681 tacgccagct gcgcgctcgc tcgctcactg aggccgggcg acctttggtc gcccggcctc
     1741 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta
     1801 gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga agatctctgc
     1861 agagggccct gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg gggtgggggt
     1921 gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat tccccaaatt
     1981 gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg
     2041 ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc
     2101 ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc ccttcccggc
     2161 caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc gaggcgcgag
     2221 ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag
     2281 tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgaa ttcaagctga
     2341 tagcaaggat cctctagagt cgactccgga ataacttcgt ataggatact ttatacgaag
     2401 ttatgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct cttaataact
     2461 tcgtatagca tacattatac gaagttatgg cgcgccttaa gcgtaatctg gaacatcgta
     2521 tgggtagaat ctggtggctt cagggaagtc cgtggcacag ctgatgaagc tgtcaatgct
     2581 ggacatgctc cgcatgtcga tgacgccctc cgtgcgtgcg ggcagagggg ccacggggct
     2641 gggcatgctg ctcatctcca gctcctctgg aggcttgctc tgcggggcca tctccttggt
     2701 gttgaggatg ggctgggact gcacgttgat gtcgatctgg tccaggggga tgtactcgcc
     2761 ctcgctggtg atcctgctct tggcggccgc agacagtctc cgcaggcgct cgccattgct
     2821 gctcagcctc atatccagct ccagcttctg gaactcatcc tcgtcgtgtc ccagctgctc
     2881 cagcagccga aacagctcct catcataggt ctttctgaac cgggccaggg caaagccgta
     2941 acaggacttg gccacgatgt cgcacacgca gtgggccacc tctgtccagt tatcgtcgat
     3001 ccagccgaag cctctagggc agatggccca caccagggga aacagaatcc acagcatgga
     3061 gaaggtaaac acggcgaact tcagcacgtg caggctgtgc tcggcgcctg tatcctgggc
     3121 cagctgggcc agccggctat actgcttggc cacgatgctc atcacgatag agtaggcgaa
     3181 gataaaccag aaacagccaa agccatacca ggcgtaggcg ccattcctca gcctagggtc
     3241 gccctctgcg tagaactcgg ccagcacgcc agacatcagg atggcaaaga tgatggcgct
     3301 acaagacacg cgataggggg ccctcagctg atacagcaga tcgtacacca gcattgggca
     3361 ggtcagcatg tagtcggcat acctgtaatt gccgaagccg atgatgaagg ggcccacatc
     3421 gccgaagttg tggcccagtg tgtcggccac gatcttgccg aagtagctca gcatggcaaa
     3481 gaaattcaca aacagggcca ggttctgctc ctcctgatag ccggcgtaaa aggaggttct
     3541 cttgctaggg ccccgagatg tccaggactc atagcccagg tagaatgtgg tgattgtcag
     3601 cacgataaag caagaccagt gggcgccgat ggccacggag ttcagcttgg tatccagata
     3661 ccagccgtcc attgtgccca cgtagaacat ctggtcggtg cctggtgcgt gtgccatggg
     3721 accggggttt tcttccacgt ctcctgcttg ctttaacaga gagaagttcg tggctccgga
     3781 gccactagtc ttcgctgtca tcatttgtac aaactcttcg tagtttacct gaccatcccc
     3841 atcgatgtct gcttccctga tcatttcatc aacctcttca tctgttaact tctctccaag
     3901 gtttgtcatc acgtggcgaa gctctgctgc actgatgtag ccattgccat ccttatcaaa
     3961 cacaccgaac gcttctctaa tttcttcttc cgtgtccctg tagctccctt ttcttgccat
     4021 cattgtcagg aactcaggga agtcgattgt gccgtcaccg tcggcatcta cttcattgat
     4081 catgtcctgc agctctgctt ctgtggggtt ctgccccaga gaccgcatca ccgtccccag
     4141 ctccttggtt gttattgtcc catccccgtc cttgtcaaat agggagaaag cctctttaaa
     4201 ttctgcgatc tgctcttcag tcagttggtc cggcaggttg tactccagct tgtgccccag
     4261 gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc
     4321 gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt
     4381 gcgctcctgg atgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg
     4441 gtcggggtag cggctgaagc actgcacgcc gtaggtcagg gtggtcacga gggtgggcca
     4501 gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc
     4561 atcgccctca ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc
     4621 gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tgctccctcc
     4681 ggtaccgccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     4741 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttc gaaagtttgg actgcacgct
     4801 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     4861 gtggtaggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga agttcgcctt
     4921 gatgccgttc ttctgcttgt cggccttgat atagacgttc tcgagtgagc tcagccgacc
     4981 tatagctctg actgcgtgac ctgtcttatt ccacttacga cgtgatgagt ccaccatggt
     5041 ggcgctagca taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa
     5101 ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg
     5161 aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     5221 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5281 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5341 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5401 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5461 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5521 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5581 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     5641 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     5701 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     5761 cctttgggcc gcctccccgc atcgataccg tcgacctcga cccgggcggc cgcttcgagc
     5821 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5881 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5941 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     6001 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
     6061 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
     6121 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     6181 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     6241 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     6301 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     6361 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6421 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6481 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6541 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6601 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6661 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6721 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6781 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6841 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6901 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6961 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     7021 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaa
//
