LOCUS       sequence-317050-        7223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of Jaws-KGC-tdTomato-ER2 under the CAG 
            promoter, in floxed/reversed (Cre-dependent) manner..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7223)
  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 7223)
  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: pAAV-CAG-FLEX-rc [Jaws-KGC-tdTomato-ER2]
FEATURES             Location/Qualifiers
     source          1..7223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     395..417
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     479..498
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(531..564)
                     /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(615..648)
                     /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    3039..3072
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3123..3156
                     /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)."
     misc_feature    3162..3750
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3215..3235)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3633..3644)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3758..3982
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3984..4124
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3984..4113
                     /label=AAV2 ITR
     rep_origin      4199..4654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4286..4305)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4496..4517
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4671..4690)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4790..4812
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4850..4868)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4936..5040
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5041..5109,5110..5901)
                     /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(5259..5278)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6072..6660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6561..6580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6722..6851
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
       61 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      121 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      181 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      241 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      301 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
      361 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
      421 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
      481 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattgaattc ataacttcgt
      541 atagcataca ttatacgaag ttattctttg caccattcta aagaataaca gtgataattt
      601 ctgggttaag gcaaataact tcgtatagga tactttatac gaagttatgc tagctcatta
      661 cacttcattc tcgtagcaga acttatacag ctcatccatg ccgtacagaa acagatggtg
      721 gcgtccttca gacctctcat actgttccac aatggtgtag tcttcattgt gggatgtgat
      781 atccagcttg gtgtccacat agtaatagcc gggcagctgc actggtttct tggccatgta
      841 gatggttttg aactccacca ggtagtggcc gccgtctttc agcttcaggg cctggtggat
      901 ttcgcccttc agcactccgt ctctaggata cagccgttca gtgctggcct cccaccccat
      961 tgttttcttc tgcatcacgg gcccatcggg tggaaaattg gtgccgcgca ttttgacttt
     1021 gtagatcaga gtgccatcct gcagagagga gtcctgtgtc acggtcacca ggcctccatc
     1081 ttcaaaattc atcactctct cccacttaaa gccctctggg aaagacagtt tcttgtaatc
     1141 aggaatgtca gcgggatgct tgacataggc cttacttccg tacataaact gaggagacag
     1201 gatatcccaa gcgaaaggca gtggtcctcc cttggtcact ttcagcttgg cagtctgtgt
     1261 gccctcgtaa ggcctccctt cgccctctcc ttcaatctca aattcgtgcc cattcatgct
     1321 gccctccatg cgcaccttaa acctcatgaa ctctttgatc acagccatat tgttatcctc
     1381 gcttgaggcg gttccactag accctgagcc ggtacttcca gtcccatggc ccaggaacag
     1441 atggtgtctc ccctcgctcc gttcatactg ctccacaata gtgtagtcct cgttgtgtga
     1501 agtgatatcc agttttgtgt ccacgtaata gtagcctggc agctgcacag gtttcttggc
     1561 catgtagatt gtcttgaact ccaccaggta atgtccccca tctttcagct tcagagcctg
     1621 gtggatttct cctttcagca ccccgtcgcg tggatacagc ctttcggtgg aggcctccca
     1681 tcccatagtt ttcttctgca tcacaggtcc atcaggggga aaatttgtcc ctctcatttt
     1741 caccttgtag atcagggtcc catcctgcag ggagctgtcc tgggtcacag tcaccaggcc
     1801 tccgtcctcg aagttcatca ctctttccca cttaaacccc tcggggaagc tcagtttctt
     1861 ataatcggga atgtcagctg gatgtttcac ataggccttg gagccgtaca taaactgtgg
     1921 gctcaggatg tcccaagcga atggcagagg cccgcccttt gtcactttca gcttggctgt
     1981 ctgggtgccc tcgtagggcc gtccctctcc ttctccctca atttcgaact cgtggccgtt
     2041 catgcttcct tccattctca ccttaaaccg catgaactct ttgatcactt cctcgccctt
     2101 actcactgcc cctgcaggtg caacattgat gtctatttga tccagaggga tatactcccc
     2161 ttcagaagta attctggatt ttactaccgg tcgatcgtcg gctggagcag ctcctccgga
     2221 cccgattccc attccactcc cagacactgt gccttcattg gcagccaccc atctcagcag
     2281 cagaaaggcg aacacgtatt tagccaggat gtccagtccg gagtatcccc agctggtcac
     2341 tcccacactc agcagggcca ctccctcaga gcccagagcg aacaggatag ggtaccccag
     2401 ccacagcacc acggtcagaa tcctcagagt gccaaagatt tcactggtcc cagcagcctc
     2461 agcatcagct ggccactgca ccagcagcac atacagcacg gccacaaaga aagcacaaga
     2521 aattccgtag aacacccagc gcagcagatg ggagctagtg atcagggcag cggccagtcc
     2581 tgtcacgcac atgccaatgt ccattgtgat ggcggtgaac aggctggcaa tatcggtgtc
     2641 ggccagcagt cccagagcca gcaggatcat gggagtggag aaagtccatg tcagataccg
     2701 gccccatggg ctcagcacct cctgtccggc cagagcgtgt ccaggtggca tctgcaggaa
     2761 gcccacagtc agcccactgg ccagtccagc gtaactagaa atagacacca gtggcaccag
     2821 cattgtggcc acccagatca gcttagctct aggggattcc agatccctcc ccatggccac
     2881 aaacagcagg atgaccactc cggccagagc aatgttcacc cagatggagc tattcagcag
     2941 gaagttggac tggatctcct gcagcacgtc gctctgtgtg gcctgcagca cggtagtggc
     3001 tgtggtgctc acggcggtca tggtggcggc ggcgcgccat aacttcgtat aatgtatgct
     3061 atacgaagtt attaagaggt ttcatattgc taatagcagc tacaatccag ctaccattct
     3121 gcataacttc gtataaagta tcctatacga agttattccg gaatcaacct ctggattaca
     3181 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     3241 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     3301 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     3361 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3421 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3481 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3541 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     3601 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3661 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3721 gtcggatctc cctttgggcc gcctccccgc ctggatcctg tgccttctag ttgccagcca
     3781 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3841 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3901 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3961 ggggatgcgg tgggctctat ggcaggaacc cctagtgatg gagttggcca ctccctctct
     4021 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     4081 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     4141 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     4201 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4261 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4321 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4381 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     4441 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4501 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     4561 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4621 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     4681 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     4741 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4801 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     4861 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4921 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4981 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5041 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5101 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5161 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5221 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5281 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5341 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5401 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5461 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5521 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5581 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5641 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5701 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5761 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5821 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5881 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5941 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6001 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6061 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6181 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6721 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
     6781 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
     6841 taggggttcc tgtcgaccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     6901 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     6961 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     7021 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     7081 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
     7141 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
     7201 cagcgatggg ggcggggggg ggg
//
