LOCUS       sequence-386785-        7264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of Jaws-KGC-tdTomato-ER2 under the CAG 
            promoter, in FLPX/reversed (Frt-dependent) manner..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7264)
  AUTHORS   Chuong AS, Miri ML, Busskamp V, Matthews GA, Acker LC, Sorensen AT, 
            Young A, Klapoetke NC, Henninger MA, Kodandaramaiah SB, Ogawa M, 
            Ramanlal SB, Bandler RC, Allen BD, Forest CR, Chow BY, Han X, Lin Y,
            Tye KM, Roska B, Cardin JA, Boyden ES
  TITLE     Noninvasive optical inhibition with a red-shifted microbial 
            rhodopsin.
  JOURNAL   Nat Neurosci. 2014 Aug;17(8):1123-9. doi: 10.1038/nn.3752. Epub 2014
            Jul 6.
  PUBMED    24997763
REFERENCE   2  (bases 1 to 7264)
  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"; doi: "10.1038/nn.3752"; 
            journalName: "Nat Neurosci"; date: "2014-08"; volume: "17"; issue: 
            "8"; pages: "1123-9"
COMMENT     Sequence Label: AAV-CAG-FLPX-rc [Jaws-KGC-tdTomato-ER2]
FEATURES             Location/Qualifiers
     source          1..7264
                     /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        387..664
                     /label=chicken beta-actin promoter
     primer_bind     943..965
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1027..1046
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1079..1112
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(3582..3615)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3705..4293
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3758..3778)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4176..4187)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4301..4525
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4527..4667
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4527..4656
                     /label=AAV2 ITR
     rep_origin      4742..5197
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4829..4848)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5039..5060
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5214..5233)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5333..5355
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5393..5411)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5479..5583
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5584..5652,5653..6444)
                     /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(5802..5821)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6615..7203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7104..7123
                     /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 gtcgacccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggggggsg sgcgccrggs ggggcggggc ggggsgrggg
      541 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
      601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
      661 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
      721 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
      781 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
      841 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggctgtc
      901 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
      961 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1021 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgaattcga
     1081 agttcctatt ctctagaaag tataggaact tcgcagaatg gtagctggat tgtagctgct
     1141 attagcaata tgaaacctct tagaagttcc tattcttcaa aaggtatagg aacttcgcta
     1201 gctcacactt cattctcgta gcagaactta tacagctcat ccatgccgta cagaaacaga
     1261 tggtggcgtc cttcagacct ctcatactgt tccacaatgg tgtagtcttc attgtgggat
     1321 gtgatatcca gcttggtgtc cacatagtaa tagccgggca gctgcactgg tttcttggcc
     1381 atgtagatgg ttttgaactc caccaggtag tggccgccgt ctttcagctt cagggcctgg
     1441 tggatttcgc ccttcagcac tccgtctcta ggatacagcc gttcagtgct ggcctcccac
     1501 cccattgttt tcttctgcat cacgggccca tcgggtggaa aattggtgcc gcgcattttg
     1561 actttgtaga tcagagtgcc atcctgcaga gaggagtcct gtgtcacggt caccaggcct
     1621 ccatcttcaa aattcatcac tctctcccac ttaaagccct ctgggaaaga cagtttcttg
     1681 taatcaggaa tgtcagcggg atgcttgaca taggccttac ttccgtacat aaactgagga
     1741 gacaggatat cccaagcgaa aggcagtggt cctcccttgg tcactttcag cttggcagtc
     1801 tgtgtgccct cgtaaggcct cccttcgccc tctccttcaa tctcaaattc gtgcccattc
     1861 atgctgccct ccatgcgcac cttaaacctc atgaactctt tgatcacagc catattgtta
     1921 tcctcgcttg aggcggttcc actagaccct gagccggtac ttccagtccc atggcccagg
     1981 aacagatggt gtctcccctc gctccgttca tactgctcca caatagtgta gtcctcgttg
     2041 tgtgaagtga tatccagttt tgtgtccacg taatagtagc ctggcagctg cacaggtttc
     2101 ttggccatgt agattgtctt gaactccacc aggtaatgtc ccccatcttt cagcttcaga
     2161 gcctggtgga tttctccttt cagcaccccg tcgcgtggat acagcctttc ggtggaggcc
     2221 tcccatccca tagttttctt ctgcatcaca ggtccatcag ggggaaaatt tgtccctctc
     2281 attttcacct tgtagatcag ggtcccatcc tgcagggagc tgtcctgggt cacagtcacc
     2341 aggcctccgt cctcgaagtt catcactctt tcccacttaa acccctcggg gaagctcagt
     2401 ttcttataat cgggaatgtc agctggatgt ttcacatagg ccttggagcc gtacataaac
     2461 tgtgggctca ggatgtccca agcgaatggc agaggcccgc cctttgtcac tttcagcttg
     2521 gctgtctggg tgccctcgta gggccgtccc tctccttctc cctcaatttc gaactcgtgg
     2581 ccgttcatgc ttccttccat tctcacctta aaccgcatga actctttgat cacttcctcg
     2641 cccttactca ctgcccctgc aggtgcaaca ttgatgtcta tttgatccag agggatatac
     2701 tccccttcag aagtaattct ggattttact accggtcgat cgtcggctgg agcagctcct
     2761 ccggacccga ttcccattcc actcccagac actgtgcctt cattggcagc cacccatctc
     2821 agcagcagaa aggcgaacac gtatttagcc aggatgtcca gtccggagta tccccagctg
     2881 gtcactccca cactcagcag ggccactccc tcagagccca gagcgaacag gatagggtat
     2941 cccagccaca gcaccacggt cagaatcctc agagtgccaa agatttcact ggtcccagca
     3001 gcctcagcat cagctggcca ctgcaccagc agcacataca gcacggccac aaagaaagca
     3061 caagaaattc cgtagaacac ccagcgcagc agatgggagc tagtgatcag ggcagcggcc
     3121 agtcctgtca cgcacatgcc aatgtccatt gtgatggcgg tgaacaggct ggcaatatcg
     3181 gtgtcggcca gcagtcccag agccagcagg atcatgggag tggagaaagt ccatgtcaga
     3241 taccggcccc atgggctcag cacctcctgt ccggccagag cgtgtccagg tggcatctgc
     3301 aggaagccca cagtcagccc actggccagt ccagcgtaac tagaaataga caccagtggc
     3361 accagcattg tggccaccca gatcagctta gctctagggg attccagatc cctccccatg
     3421 gccacaaaca gcaggatgac cactccggcc agagcaatgt tcacccagat ggagctattc
     3481 agcaggaagt tggactggat ctcctgcagc acgtcgctct gtgtggcctg cagcacggta
     3541 gtggctgtgg tgctcacggc ggtcatggtg gcggcggatc cgaagttcct atactttcta
     3601 gagaatagga acttcttgcc ttaacccaga aattatcact gttattcttt agaatggtgc
     3661 aaagagaagt tcctatacct tttgaagaat aggaacttct ccggaatcaa cctctggatt
     3721 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     3781 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     3841 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     3901 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     3961 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     4021 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4081 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     4141 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     4201 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     4261 cgagtcggat ctccctttgg gccgcctccc cgcctggatc ctgtgccttc tagttgccag
     4321 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     4381 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     4441 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     4501 gctggggatg cggtgggctc tatggcagga acccctagtg atggagttgg ccactccctc
     4561 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4621 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4681 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4741 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4801 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4861 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4921 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     4981 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5041 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     5101 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     5161 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     5221 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     5281 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5341 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     5401 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     5461 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5521 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5581 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5641 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5701 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5761 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5821 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5881 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5941 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6001 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6061 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6121 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6181 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6241 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6301 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6361 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6421 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6481 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6541 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6601 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6661 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6721 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6781 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6841 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6901 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6961 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7021 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7081 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7141 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7201 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7261 atgt
//
