LOCUS       sequence-485699-        6998 bp    DNA     circular SYN 10-SEP-2025
DEFINITION  Flexed - 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 6998)
  AUTHORS   Adesnik H
  TITLE     Miscellaneous Optogenetic Tools
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6998)
  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            "Unpublished"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_DIO_ChroME2f_GCaMP8m_ST
FEATURES             Location/Qualifiers
     source          1..6998
                     /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)."
     CDS             3014..3031
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     3313..3334
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3440..3461)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3701..3720)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    4489..4522
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4573..4606
                     /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(4652..5099)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   5181..5310
                     /label=AAV2 ITR
     rep_origin      5382..5837
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5469..5488)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5679..5700
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5863..5967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5968..6036,6037..6828)
                     /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(6186..6205)
                     /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 agcgctagta tggaaacagc
     2041 cgccacaatg acccacgcct ttatctcagc cgtgcctagc gccgaagcca caattagagg
     2101 cctgctgagc gccgcagcag tggtgacacc agcagcagac gctcacggag aaacctctaa
     2161 cgccacaaca gccggagccg atcacggttg cttcccccac atcaaccacg gaaccgagct
     2221 gcagcacaag atcgcagtgg gactccagtg gttcaccgtg atcgtggcta tcgtgcagct
     2281 catcttctac ggttggcaca gcttcaaggc cacaaccggc tgggaggagg tctacgtctg
     2341 cgtgatcgag ctcgtcaagt gcttcatcga gctgttccac gatgtcgaca gcccagccac
     2401 agtgtaccag accaacggag gagccgtggc ttggctgcgg tacagcgagt ggctcctgac
     2461 ttgccccgtg atcctgatcc acctgagcaa cctgaccgga ctgcacgaag agtacagcaa
     2521 gcggaccatg accatcctgg tgaccgacat cggcaacatc gtgtggggga tcacagccgc
     2581 ctttacaaag ggccccctga agatcctgtt cttcatgatc ggcctgttct acggcgtgac
     2641 ttgcttcttc cagatcgcca aggtgtatat cgagagctac cacaccctgc ccaaaggcgt
     2701 ctgccggaag atttgcaaga tcatggccta cgtcttcttc tgctcttggc tgatgttccc
     2761 cgtgatgttc atcgccggac acgagggact gggcctgatc acaccttaca ccagcggaat
     2821 cggccacctg atcctggatc tgatcgccaa gaacacttgg ggcttcctgg gccaccacct
     2881 gagagtgaag atccacgagc acatcctgat ccacggcgac atccggaaga caaccaccat
     2941 caacgtggcc ggcgagaaca tggagatcga gaccttcgtc gacgaggagg aggagggagg
     3001 agtggcggcc gcccatcatc accatcatca cacgcgtcgc aagaagacct tcaaggaggt
     3061 ggccaccgcc gtgaagatca tcggccgcct gatgggcctg aagatcaacg tctatatcaa
     3121 ggccgacaag cagaagaacg gcatcaaggc gaacttccac atccgccaca acatcgagga
     3181 cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt
     3241 gctgctgccc gacaaccact acctgagcgt ggagtccaaa ctttcgaaag accccaacga
     3301 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3361 ggacgagctg tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt
     3421 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3481 cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3541 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3601 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3661 gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3721 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3781 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacc tgccggacca
     3841 actgactgaa gagcagatcg cagaatacaa agaggctttc tccctatttg acaaggacgg
     3901 ggatgggaca ataacaacca aggagctggg gacggtgatg cggtctctgg ggcagaaccc
     3961 cacagaagca gagctgcagg acatgatcaa tgaagtagat gccgacggtg acggcacaat
     4021 cgacttccct gagttcctga caatgatggc aagaaaaatg aaatacaggg acacggaaga
     4081 agaaattaga gaagcgttcg gtgtgtttga taaggatggc aatggctaca tcagtgcagc
     4141 agagcttcgc cacgtgatga caaaccttgg agagaagtta acagatgaag aggttgatga
     4201 aatgatcagg gaagcagaca tcgatgggga tggtcaggta aactacgaag agtttgtaca
     4261 aatgatgaca gcgaagcaat tgcagtccca gcccatcctc aacaccaagg agatggcccc
     4321 gcagagcaag cctccagagg agctggagat gagcagcatg cccagccccg tggcccctct
     4381 gcccgcacgc acggagggcg tcatcgacat gcggagcatg tccagcattg acagcttcat
     4441 cagctgtgcc acggacttcc ctgaagccac cagattctaa ggcgcgccat aacttcgtat
     4501 aatgtatgct atacgaagtt attaagaggt ttcatattgc taatagcagc tacaatccag
     4561 ctaccattct gcataacttc gtataaagta tcctatacga agttattccg gagtcgactc
     4621 tagaggatcc ttgctatcag cttgaattcg actgcgctct caggcacgac acgactcctc
     4681 cgctgcccac cgcagactga ggcagcgctg agtcgccggc gccgcagcgc agatggtcgc
     4741 gcccgtgccc ccctatctcg cgcctcgcgt ggtgcggtcc ggctgggccg gcggcggcgc
     4801 ggacgcgacc aaggtggccg ggaaggggag tttgcggggg accggcgagt gacgtcagcg
     4861 cgccttcagt gctgaggcgg cggtggcgcg cgccgccagg cgggggcgaa ggcactgtcc
     4921 gcggtgctga agctggcagt gcgcacgcgc ctcgccgcat cctgtttccc ctccccctct
     4981 ctgatagggg atgcgcaatt tggggaatgg gggttgggtg cttgtccagt gggtcggggt
     5041 cggtcgtcag gtaggcaccc ccaccccgcc tcatcctggt cctaaaaccc acttgcactc
     5101 atacgcaggg ccctctgcag agatcttcct agagcatggc tacgtagata agtagcatgg
     5161 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     5221 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     5281 ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga
     5341 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     5401 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5461 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5521 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5581 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5641 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5701 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5761 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5821 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5881 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5941 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6001 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6061 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6121 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6181 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6241 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6301 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6361 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6421 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6481 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6541 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6601 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6661 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6721 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6781 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6841 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6901 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6961 tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
