LOCUS       sequence-455103-        7398 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 7398)
  AUTHORS   Adesnik H
  TITLE     Miscellaneous Optogenetic Tools
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7398)
  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            "Unpublished"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..7398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   379..508
                     /label=AAV2 ITR
     promoter        590..1037
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(1083..1116)
                     /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(1167..1200)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1212..1238)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2430..2486)
                     /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     3058..3077
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3317..3338
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3444..3465)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3678..3734)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     regulatory      3746..3755
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3762..3795
                     /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    3846..3879
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3904..4492
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3957..3977)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4375..4386)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4495..4511)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4496..4512)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4541..4662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4552..4571)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4606..4625
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4824..4953
                     /label=AAV2 ITR
     rep_origin      5133..5588
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5220..5239)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5430..5451
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5614..5718
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5719..5787,5788..6579)
                     /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(5937..5956)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6750..7338
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7239..7258
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgc
      181 gcgctcgctc gctcrmctgg rmmgcssgsr swragcscrr cgcrwtkrrt gkgmgttrgc
      241 wcrykcakyr gkcacscyrs gcktwacacc wcaccckycs sgcttaatgc gccgctacag
      301 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      361 tcttcgctat tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaangccc
      421 gggngncggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      481 gtggccaact ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta
      541 tctacgtagc catgctctag gaagatctct gcagagggcc ctgcgtatga gtgcaagtgg
      601 gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac
      661 tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg
      721 gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt
      781 cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac
      841 tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg
      901 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
      961 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
     1021 cgtgcctgag agcgcagtcg aattcaagct gatagcaagg atcctctaga gtcgactccg
     1081 gaataacttc gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc
     1141 tgctattagc aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat
     1201 ggcgcgcctt aagcgtaatc tggaacatcg tatgggtaga atctggtggc ttcagggaag
     1261 tccgtggcac agctgatgaa gctgtcaatg ctggacatgc tccgcatgtc gatgacgccc
     1321 tccgtgcgtg cgggcagagg ggccacgggg ctgggcatgc tgctcatctc cagctcctct
     1381 ggaggcttgc tctgcggggc catctccttg gtgttgagga tgggctggga ctgcacgttg
     1441 atgtcgatct ggtccagggg gatgtactcg ccctcgctgg tgatcctgct cttggcggcc
     1501 gcagacagtc tccgcaggcg ctcgccattg ctgctcagcc tcatatccag ctccagcttc
     1561 tggaactcat cctcgtcgtg tcccagctgc tccagcagcc gaaacagctc ctcatcatag
     1621 gtctttctga accgggccag ggcaaagccg taacaggact tggccacgat gtcgcacacg
     1681 cagtgggcca cctctgtcca gttatcgtcg atccagccga agcctctagg gcagatggcc
     1741 cacaccaggg gaaacagaat ccacagcatg gagaaggtaa acacggcgaa cttcagcacg
     1801 tgcaggctgt gctcggcgcc tgtatcctgg gccagctggg ccagccggct atactgcttg
     1861 gccacgatgc tcatcacgat agagtaggcg aagataaacc agaaacagcc aaagccatac
     1921 caggcgtagg cgccattcct cagcctaggg tcgccctctg cgtagaactc ggccagcacg
     1981 ccagacatca ggatggcaaa gatgatggcg ctacaagaca cgcgataggg ggccctcagc
     2041 tgatacagca gatcgtacac cagcattggg caggtcagca tgtagtcggc atacctgtaa
     2101 ttgccgaagc cgatgatgaa ggggcccaca tcgccgaagt tgtggcccag tgtgtcggcc
     2161 acgatcttgc cgaagtagct cagcatggca aagaaattca caaacagggc caggttctgc
     2221 tcctcctgat agccggcgta aaaggaggtt ctcttgctag ggccccgaga tgtccaggac
     2281 tcatagccca ggtagaatgt ggtgattgtc agcacgataa agcaagacca gtgggcgccg
     2341 atggccacgg agttcagctt ggtatccaga taccagccgt ccattgtgcc cacgtagaac
     2401 atctggtcgg tgcctggtgc gtgtgccatg ggaccggggt tttcttccac gtctcctgct
     2461 tgctttaaca gagagaagtt cgtggctccg gagccactag tcttcgctgt catcatttgt
     2521 acaaactctt cgtagtttac ctgaccatcc ccatcgatgt ctgcttccct gatcatttca
     2581 tcaacctctt catctgttaa cttctctcca aggtttgtca tcacgtggcg aagctctgct
     2641 gcactgatgt agccattgcc atccttatca aacacaccga acgcttctct aatttcttct
     2701 tccgtgtccc tgtagctccc ttttcttgcc atcattgtca ggaactcagg gaagtcgatt
     2761 gtgccgtcac cgtcggcatc tacttcattg atcatgtcct gcagctctgc ttctgtgggg
     2821 ttctgcccca gagaccgcat caccgtcccc agctccttgg ttgttattgt cccatccccg
     2881 tccttgtcaa atagggagaa agcctcttta aattctgcga tctgctcttc agtcagttgg
     2941 tccggcaggt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     3001 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     3061 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggatgtagcc ttcgggcatg
     3121 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     3181 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     3241 atgaacttca gggtcagctt gccgtaggtg gcatcgccct caccctcgcc ggacacgctg
     3301 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     3361 agctcctcgc ccttgctcac catgctccct ccggtaccgc ccttgtacag ctcgtccatg
     3421 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     3481 ttggggtctt tcgaaagttt ggactgcacg ctcaggtagt ggttgtcggg cagcagcacg
     3541 gggccgtcgc cgatgggggt gttctgctgg tagtggtagg cgagctgcac gccgccgtcc
     3601 tcgatgttgt ggcggatctt gaagttcgcc ttgatgccgt tcttctgctt gtcggccttg
     3661 atatagacgt tctcgagtga gctcagccga cctatagctc tgactgcgtg acctgtctta
     3721 ttccacttac gacgtgatga gtccaccatg gtggcgctag cataacttcg tataaagtat
     3781 cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc
     3841 aaagaataac ttcgtataat gtatgctata cgaagttatg aattcgatat caagcttatc
     3901 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3961 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4021 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     4081 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     4141 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     4201 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4261 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     4321 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4381 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4441 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     4501 cgtcgacctc gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt
     4561 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     4621 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     4681 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc
     4741 tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca
     4801 tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct
     4861 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4921 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg
     4981 gggagaggcg gtttgcgtat tgggcgctct tccscttcct csctcrctsa ctcgcynckc
     5041 wskgwsskws sgmksgcgca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5101 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     5161 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     5221 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5281 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5341 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5401 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5461 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5521 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5581 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5641 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5701 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5761 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5821 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5881 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5941 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6001 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6061 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6121 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6181 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6241 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6301 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6361 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6421 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6481 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6541 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6601 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6661 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6721 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6781 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6841 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6901 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6961 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7021 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7081 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7141 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7201 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7261 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7321 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7381 tggccttttg ctcacatg
//
