LOCUS       sequence-195127-        7225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent viral expression of ArchT-tdTomato for neuronal 
            inhibition.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7225)
  AUTHORS   Han X, Chow BY, Zhou H, Klapoetke NC, Chuong A, Rajimehr R, Yang A, 
            Baratta MV, Winkle J, Desimone R, Boyden ES
  TITLE     A high-light sensitivity optical neural silencer: development and 
            application to optogenetic control of non-human primate cortex.
  JOURNAL   Front Syst Neurosci. 2011;5:18. Epub 2011 Apr 13.
  PUBMED    21811444
REFERENCE   2  (bases 1 to 7225)
  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: "Front Syst 
            Neurosci."; date: "2011"; volume: "5"; pages: "18. Epub 2011 Apr 13"
COMMENT     Sequence Label: pAAV-FLEX-ArchT-tdTomato
FEATURES             Location/Qualifiers
     source          1..7225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        135..412
                     /label=chicken beta-actin promoter
     primer_bind     691..713
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     775..794
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    829..862
                     /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    925..958
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(981..2408)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN
                     FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL
                     KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER
                     SEGRHHLFLYGMDELYK"
     regulatory      3158..3167
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3184..3217)
                     /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(3288..3321)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3363..3951
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3416..3436)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3834..3845)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3994..4115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4005..4024)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4059..4078
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4277..4406
                     /label=AAV2 ITR
     rep_origin      complement(4643..5231)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4723..4742)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5402..6193,6194..6262))
                     /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     6025..6044
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6263..6367)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6393..6848)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6530..6551)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6742..6761
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6920..7049
                     /label=AAV2 ITR
ORIGIN
        1 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
       61 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      121 cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc
      181 ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc
      241 gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgrggggc ggggcggggc
      301 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
      361 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
      421 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
      481 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
      541 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
      601 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggctgtccg cggggggacg
      661 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
      721 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
      781 tgctggttat tgtgctgtct catcattttg gcaaagaatt ggatccccat aacttcgtat
      841 aaagtatcct atacgaagtt atatcaaaat aggaagacca atgcttcacc atcgacccga
      901 attgccaagc atcaccatcg acccataact tcgtataatg tatgctatac gaagttatac
      961 tagggatccc ccctcgagtt ttacttgtac agctcgtcca tgccgtacag gaacaggtgg
     1021 tggcggccct cggagcgctc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg
     1081 atgtccagct tggtgtccac gtagtagtag ccgggcagtt gcacgggctt cttggccatg
     1141 tagatggtct tgaactccac caggtagtgg ccgccgtcct tcagcttcag ggcctggtgg
     1201 atctcgccct tcagcacgcc gtcgcggggg tacaggcgct cggtggaggc ctcccagccc
     1261 atggtcttct tctgcattac ggggccgtcg ggggggaagt tggtgccgcg catcttcacc
     1321 ttgtagatca gcgtgccgtc ctgcagggag gagtcctggg tcacggtcac cagaccgccg
     1381 tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcttgtaa
     1441 tcggggatgt cggcggggtg cttcacgtac gccttggagc cgtacatgaa ctggggggac
     1501 aggatgtccc aggcgaaggg cagggggccg cccttggtca ccttcagctt ggcggtctgg
     1561 gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcatg
     1621 gagccctcca tgcgcacctt gaagcgcatg aactctttga tgacggccat gttgttgtcc
     1681 tcggaggagg cggtgccgga gctgccgctg ccggtgctgc cggtgccatg ccccaggaac
     1741 aggtggtggc ggccctcgga gcgctcgtac tgttccacga tggtgtagtc ctcgttgtgg
     1801 gaggtgatgt ccagcttggt gtccacgtag tagtagccgg gcagttgcac gggcttcttg
     1861 gccatgtaga tggtcttgaa ctccaccagg tagtggccgc cgtccttcag cttcagggcc
     1921 tggtggatct cgcccttcag cacgccgtcg cgggggtaca ggcgctcggt ggaggcctcc
     1981 cagcccatgg tcttcttctg cattacgggg ccgtcggggg ggaagttggt gccgcgcatc
     2041 ttcaccttgt agatcagcgt gccgtcctgc agggaggagt cctgggtcac ggtcaccaga
     2101 ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa ggacagcttc
     2161 ttgtaatcgg ggatgtcggc ggggtgcttc acgtacgcct tggagccgta catgaactgg
     2221 ggggacagga tgtcccaggc gaagggcagg gggccgccct tggtcacctt cagcttggcg
     2281 gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg
     2341 ttcatggagc cctccatgcg caccttgaag cgcatgaact ctttgatgac ctcctcgccc
     2401 ttgctcacta ccggtcgggg ctcgggggcc tcggtgtcgc ccaggatggc cctgctcctc
     2461 agcaggatga agccgaagcc cactttggcg gtcacgtcca gcaccatgaa cagcagggtc
     2521 tcgattccca gtcccaccac tccagctccc tcggttccga tgatccacag gatggggtag
     2581 gcggtccaca gcaccagcac cagggcggtc agggtgttga aggtgctagc cacctctggt
     2641 cccctctcct tagcagcagc cctcaggctg gtagccagga agtacagcac cacgatcatg
     2701 cagatggtgc tgaacagcca ccagctgtac ctagccagtg gggtgtggct cagggcgccg
     2761 atcaggccgg tcacgatcat cagggcgtcc actcccacca gggttccgat gctcaccctg
     2821 tccaccttag ccagcagggc caggtcgagc agaagcaaag gggtggtgaa cagccagtcg
     2881 gcgtatctgg cgtagtagat gtccagcacc tcgccggcca cggtcacctc ggtcaggccg
     2941 atgccgaaga acatgctcag gtaggcggcg ctggcgatgc cgggcaccag gattgtgatg
     3001 ctgtagtact ccctggcctc cttgtcggtc acgccccagc ccttcacgat gaagtagaag
     3061 gtgccgatca gcatcagcag ggttccgatt cccagccaca gggtctctgg ccttccgtcg
     3121 cccagcaggt cgtatcctgc ctgcagagcg atggggtcca tggtggcggt accgaattct
     3181 agtataactt cgtataggat actttatacg aagttatcat tgggattctt cctattttga
     3241 tccaagcatc accatcgacc ctctagtcca gatctcacca tcgacccata acttcgtata
     3301 gcatacatta tacgaagtta tgtccctcga agaggttcga attcgatatc aagcttatcg
     3361 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     3421 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     3481 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     3541 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     3601 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     3661 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     3721 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     3781 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     3841 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     3901 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc
     3961 gtcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     4021 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     4081 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     4141 tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat
     4201 gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta gcatggcggg
     4261 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4321 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4381 gcctcagtga gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag
     4441 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4501 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4561 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4621 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4681 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4741 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4801 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4861 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4921 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4981 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5041 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5101 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5161 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5221 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5281 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5341 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5401 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5461 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5521 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5581 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5641 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5701 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5761 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5821 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5881 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5941 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6001 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6061 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6121 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6181 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6241 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6301 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6361 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     6421 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     6481 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     6541 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6601 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     6661 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6721 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     6781 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6841 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     6901 ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
     6961 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
     7021 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
     7081 atctacgtag ccatgctcta ggaagagtac cattgacgtc aataatgacg tatgttccca
     7141 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     7201 cccacttggc agtacatcaa gtgta
//
