LOCUS       sequence-379290-        8098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-vector for bidirectional optogenetic manipulation of 
            Cre-positive neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8098)
  AUTHORS   Wiegert Lab
  TITLE     Wiegert Lab Plasmids 2022
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8098)
  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            "Unpublished"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: hSyn-DIO-somBiPOLES-eGFP
FEATURES             Location/Qualifiers
     source          1..8098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /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(744..777)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(999..2042)
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=Chrimson
                     /translation="ELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTEWW
                     CDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSIAI
                     ALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRYFE
                     WLLSCPVILIKLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIVSC
                     IYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLKLS
                     PYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEEFV
                     EEEDEDTV"
     CDS             complement(join(2403..3113,3114..3116))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             complement(join(2403..3113,3114..3116))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2445..2466)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2794..2813
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3053..3074
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    4083..4116
                     /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    4167..4200
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4225..4813
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4278..4298)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4696..4707)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4845..5321
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5044..5063)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5361..5501
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5361..5490
                     /label=AAV2 ITR
     rep_origin      5576..6031
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5663..5682)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5873..5894
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6048..6067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6167..6189
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6227..6245)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6313..6417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6418..6486,6487..7278)
                     /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(6636..6655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7449..8037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7938..7957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgagctttc tagatcagaa gcgggtggcc tcggggaagt cggtggcgca gctgatgaag
      841 ctgtcgatgc tgctcatgct gcgcatgtcg atcacgccct cggtgcgggc gggcaggggg
      901 gccacggggc tgggcatgct gctcatctcc agctcctcgg ggggcttgct ctggggggcc
      961 atctccttgg tgttcaggat gggctggctc tgctcgagca ctgtgtcctc gtcctcctcc
     1021 tccacgaact cttccacttc cacctcctcg cctccgatct ccatcttggt ggtcttccgg
     1081 atgtcgccgt ggatcaggat gtgctcgtgg atcttgatcc tcaggtggtg ggccaggaag
     1141 gtccaaaact ccttggcgat gatgtcgcag atgctgtggc cgatgctgtt ggcgtaaggg
     1201 ctcagcttca gcagtccttc tggtcccact gcccagagga ttgggtagct gccccaagag
     1261 gcgaagtaag cgtaggccat cagcttcacg accatgcggc aatggccttt aggcacgctg
     1321 tggttggctt ccacgtagca cttggcggcc tggaagtaca tgtagccgcc gtagatgcaa
     1381 gacacgatat acagcagcca cttgagccaa tcggtagcca gtcctgcggc catgccgaac
     1441 acgatcattc ccacgcaaga cacgatcagg cccatggtcc gcttgctgta gtcgttcttc
     1501 aggccgctca ggttgctcag cttgatcagg atcacggggc aagacagcag ccactcgaag
     1561 tagcgcaggc aataggcgtg gttgccggta gacaggtaca ctgtggcggg gctgctgaac
     1621 tccttgaaga tctccagggt cacgaacagc acctcgacgc aacagacgta gacctcctcc
     1681 caaccgcaag tggccttcca agcgctgaag ccgtagaatg tcagcagggc gatggcgatg
     1741 ctgaaagcaa tccactggca gacctgggcg ccgatctttt ctcctggtgt gcctctgctg
     1801 tgcagccaag cctgcttctt ctccactccc acgaccaggt agccgcctgt ggcctccacg
     1861 aagcaatatc cgtatccggc gtcgctcagt ccgtaagaag ggtcacacca ccactcggtg
     1921 ctgaagcact cgcctgtagg tgtcattcct ccacagccca tgtcctcgtg gtggtagggg
     1981 ttaggccaag ggttgatgcc tccggcggca aacagagatc tggtggcgct gctgatcagc
     2041 tcttgtacag ctccctccgg acgtccggtg atctgggcct gtctagagct gttttcagag
     2101 atgttgtagg aaatctgcag ccctctttcc ccatacacat ccggtctcaa ggttaccccc
     2161 ggtgacttta actggtcctg gtagtcgggg gtgtagggat caatggtctg catcagcaca
     2221 tagatgcaca aggcaaagag cccagtcatg accacgtaga aagctgcata gtacaggctg
     2281 atccacaccc accgggctgg ggtgcggccc agcatctgcc cagtgtccgg gttccaacag
     2341 tattgccgga attcggccat gcgctggctg catgacttct tctcctgcag ggctgcaccg
     2401 gtcttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     2461 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag
     2521 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     2581 gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     2641 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     2701 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     2761 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     2821 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     2881 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     2941 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     3001 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     3061 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcacgggt
     3121 accgccacgt tgatgtcgat ctggtccagg gggatgtact cgccctcgct ggtgatcctg
     3181 ctcttggccg ccgtacgtcc ccttccaaac attctgatat tgacttcgtc gttaggaggg
     3241 gcattcttct cgtgattcag gccaaaagga tcgtcggctt cgggtttctc ctgccctggc
     3301 agacatgcgg ggtcccactt cccttccagc actccccagg cagtgctcca caggatcact
     3361 ccatatgtat ttttgcacag gccgtcagcc agcaggtaca gaatggcgct cacgttctcg
     3421 tggatcacgc agactccggt gggtgaaatc agccacacga ttgggaacat agtccaagca
     3481 ataaagaaga tcaggcgcag catcttgatc ctcaccacca ccttctgtgc cagaggtgtc
     3541 ccgacagatt caaagccata cagccccact tgaaagattg tgaagaccac actgtactcg
     3601 aacaccaggc aggtggcccc aaagaaaaag aacagccact tcatatactc tgcgggagaa
     3661 attgtagcgg tgattcccat gaccactcta atgatggaag cggcctgtgc aatgggattc
     3721 agtgggatac ttttgtactt caccagagcc atgttagaaa tctgtcccag catgattgga
     3781 caggtgcaca gccaggaggc cattcggctc caaggaaaga atctgccgtc cctcatctgc
     3841 agggttccgt tgccagtact ggccagtgca taagtgatag attcgacaaa gggcaggtac
     3901 acggactccc aagggacctt gggagccctg ctaatcatga agattgtctg gcagcctgag
     3961 accacggccc acaccagctg cagcagtgag acgaaggtgg agtcgattct ggacatattg
     4021 tagcactcgg tgtgagttga tgcccattcc ccatagacga cctgtgatgc catggatcta
     4081 gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac
     4141 tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat
     4201 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     4261 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4321 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4381 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4441 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4501 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4561 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4621 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     4681 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4741 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4801 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg
     4861 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     4921 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     4981 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     5041 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     5101 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     5161 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     5221 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     5281 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     5341 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     5401 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     5461 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     5521 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     5581 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5641 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5701 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5761 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     5821 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5881 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     5941 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6001 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     6061 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     6121 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     6181 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     6241 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     6301 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     6361 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     6421 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     6481 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     6541 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     6601 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     6661 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     6721 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     6781 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     6841 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6901 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6961 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7021 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7081 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     7141 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7201 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7261 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7321 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7381 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     7441 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     7501 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     7561 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     7621 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     7681 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     7741 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     7801 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     7861 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7921 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7981 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     8041 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
