LOCUS       sequence-458443-        7100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre off - 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 7100)
  AUTHORS   Adesnik H
  TITLE     Miscellaneous Optogenetic Tools
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7100)
  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            "Unpublished"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-hSyn-DO-ChroME2f-ST-P2A-GCaMP8s
FEATURES             Location/Qualifiers
     source          1..7100
                     /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"
     protein_bind    complement(1900..1933)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1984..2017)
                     /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)."
     primer_bind     2583..2602
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2842..2863
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2969..2990)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3272..3289)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3299..3355)
                     /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             complement(3566..3589)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    4591..4624
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4675..4708
                     /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)."
     promoter        complement(4754..5201)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   5283..5412
                     /label=AAV2 ITR
     rep_origin      5484..5939
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5571..5590)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5781..5802
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5965..6069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6070..6138,6139..6930)
                     /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(6288..6307)
                     /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 atcgaattca taacttcgta tagcatacat
     1921 tatacgaagt tattctttgc accattctaa agaataacag tgataatttc tgggttaagg
     1981 caaataactt cgtataggat actttatacg aagttatgct agcttacttc gctgtcatca
     2041 tttgtacaaa ctcttcgtag tttacctgac catccccatc gatgtctgct tccctgatca
     2101 tttcatcaac ctcttcatct gttaacttct ctccaaggtt tgtcatcacg tggcgaagct
     2161 ctgctgcact gatgtagcca ttgccatcct tatcaaacac accgaacgct tctctaattt
     2221 cttcttccgt gtccctgtat ttcatttttc ttgccatcat tgtcaggaac tcagggaagt
     2281 cgattgtgcc gtcaccgtcg gcatctactt cattgatcat gtcctgcagc tctgcttctg
     2341 tggggttgtg tccgagagac cgcatcaccg tccccagctc cttggttgtt attgtcccat
     2401 ccccgtcctt gtcaaatagg gagaaagcct ctttgtattc tgcgatctgc tcttcagtca
     2461 gttggtccgg caggttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt
     2521 cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc
     2581 gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggatg tagccttcgg
     2641 gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact
     2701 gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg
     2761 tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcaccc tcgccggaca
     2821 cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg
     2881 tgaacagctc ctcgcccttg ctcaccatgc tccctccggt accgcccttg tacagctcgt
     2941 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     3001 tctcgttggg gtctttcgaa agtttggact ccacgctcag gtagtggttg tcgggcagca
     3061 gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtaggcgagc tgcacgccgc
     3121 cgtcctcgat gttgtggcgg atgtggaagt tcgccttgat gccgttcttc tgcttgtcgg
     3181 ccttgatata gacgttgatc ttcaggccca tcagcatggc gatgatcttc acggcggtgg
     3241 ccacctcctt gaaggtcttc ttgcgacgcg tgtgatgatg gtgatgatgc atactagtag
     3301 gtccagggtt ctcctccacg tctccagcct gcttcagcag gctgaagtta gtagctccgc
     3361 ttcctgccgc gaatctggtg gcttcaggga agtccgtggc acagctgatg aagctgtcaa
     3421 tgctggacat gctccgcatg tcgatgacgc cctccgtgcg tgcgggcaga ggggccacgg
     3481 ggctgggcat gctgctcatc tccagctcct ctggaggctt gctctgcggg gccatctcct
     3541 tggtgttgag gatgggctgg gactgcttat cgtcatcgtc tttgtaatct accggtgcgg
     3601 ccgccactcc tccctcctcc tcctcgtcga cgaaggtctc gatctccatg ttctcgccgg
     3661 ccacgttgat ggtggttgtc ttccggatgt cgccgtggat caggatgtgc tcgtggatct
     3721 tcactctcag gtggtggccc aggaagcccc aagtgttctt ggcgatcaga tccaggatca
     3781 ggtggccgat tccgctggtg taaggtgtga tcaggcccag tccctcgtgt ccggcgatga
     3841 acatcacggg gaacatcagc caagagcaga agaagacgta ggccatgatc ttgcaaatct
     3901 tccggcagac gcctttgggc agggtgtggt agctctcgat atacaccttg gcgatctgga
     3961 agaagcaagt cacgccgtag aacaggccga tcatgaagaa caggatcttc agggggccct
     4021 ttgtaaaggc ggctgtgatc ccccacacga tgttgccgat gtcggtcacc aggatggtca
     4081 tggtccgctt gctgtactct tcgtgcagtc cggtcaggtt gctcaggtgg atcaggatca
     4141 cggggcaagt caggagccac tcgctgtacc gcagccaagc cacggctcct ccgttggtct
     4201 ggtacactgt ggctgggctg tcgacatcgt ggaacagctc gatgaagcac ttgacgagct
     4261 cgatcacgca gacgtagacc tcctcccagc cggttgtggc cttgaagctg tgccaaccgt
     4321 agaagatgag ctgcacgata gccacgatca cggtgaacca ctggagtccc actgcgatct
     4381 tgtgctgcag ctcggttccg tggttgatgt gggggaagca accgtgatcg gctccggctg
     4441 ttgtggcgtt agaggtttct ccgtgagcgt ctgctgctgg tgtcaccact gctgcggcgc
     4501 tcagcaggcc tctaattgtg gcttcggcgc taggcacggc tgagataaag gcgtgggtca
     4561 ttgtggcggc tgtttccata ctggcgcgcc ataacttcgt ataatgtatg ctatacgaag
     4621 ttattaagag gtttcatatt gctaatagca gctacaatcc agctaccatt ctgcataact
     4681 tcgtataaag tatcctatac gaagttattc cggagtcgac tctagaggat ccttgctatc
     4741 agcttgaatt cgactgcgct ctcaggcacg acacgactcc tccgctgccc accgcagact
     4801 gaggcagcgc tgagtcgccg gcgccgcagc gcagatggtc gcgcccgtgc ccccctatct
     4861 cgcgcctcgc gtggtgcggt ccggctgggc cggcggcggc gcggacgcga ccaaggtggc
     4921 cgggaagggg agtttgcggg ggaccggcga gtgacgtcag cgcgccttca gtgctgaggc
     4981 ggcggtggcg cgcgccgcca ggcgggggcg aaggcactgt ccgcggtgct gaagctggca
     5041 gtgcgcacgc gcctcgccgc atcctgtttc ccctccccct ctctgatagg ggatgcgcaa
     5101 tttggggaat gggggttggg tgcttgtcca gtgggtcggg gtcggtcgtc aggtaggcac
     5161 ccccaccccg cctcatcctg gtcctaaaac ccacttgcac tcatacgcag ggccctctgc
     5221 agagatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta
     5281 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     5341 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     5401 gcgagcgcgc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5461 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5521 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5581 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5641 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5701 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5761 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5821 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5881 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     5941 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6001 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6061 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6121 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6181 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6241 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6301 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6361 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6421 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6481 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6541 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6601 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6661 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6721 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6781 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6841 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6901 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6961 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7021 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7081 gaaaagatca aaggatcttc
//
