LOCUS       sequence-177041-        6780 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of Jaws-KGC-GFP-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 6780)
  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 6780)
  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-GFP-ER2]
FEATURES             Location/Qualifiers
     source          1..6780
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   441..570
                     /label=AAV2 ITR
     rep_origin      642..1097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(729..748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     939..960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1123..1227
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1228..1296,1297..2088)
                     /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(1446..1465)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2259..2847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2748..2767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3084..3213
                     /label=AAV2 ITR
     polyA_signal    3383..3590
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    3631..4219
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3737..3748
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4146..4166
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    4252..4285
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4356..4389
                     /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             join(5327..5329,5330..6040)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5327..5329,5330..6040)
                     /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(5369..5390)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5630..5649)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5977..5998
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(6078..6111)
                     /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(6174..6207)
                     /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)."
     primer_bind     complement(6242..6261)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(6323..6345)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(join(6624..6780,1..118))
                     /label=chicken beta-actin promoter
ORIGIN
        1 cccccccccc ccgcccccat cgctgcacaa aataattaaa aaataaataa atacaaaatt
       61 gggggtgggg agggggggga gatggggaga gtgaagcaga acgtggggct cacctcgacc
      121 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
      181 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
      241 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
      301 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
      361 gtcaatggtt aattaaccta cgatcttcct agagcatggc tacgtagata agtagcatgg
      421 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
      481 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
      541 ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga
      601 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      661 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      721 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      781 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      841 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      901 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      961 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     1021 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     1081 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1141 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1201 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1261 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1321 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1381 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1441 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1501 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1561 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1621 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1681 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1741 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1801 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1861 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1921 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1981 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2041 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2101 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2161 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2221 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2281 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2341 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2401 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2461 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2521 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2581 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2641 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2701 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2761 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2821 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2881 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2941 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3001 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     3061 cgttggccga ttcattaatg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa
     3121 agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag
     3181 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
     3241 acttatctac gtagccatgc tctaggaaga tccttatcga ttttaccaca tttgtagagg
     3301 ttttacttgc tttaaaaaac ctcccacatc tccccctgaa cctgaaacat aaaatgaatg
     3361 caattgtaca gtccggaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc
     3421 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
     3481 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
     3541 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgagcttta
     3601 tgagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
     3661 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
     3721 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
     3781 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
     3841 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
     3901 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
     3961 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
     4021 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
     4081 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     4141 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     4201 tgtaatccag aggttgatta tcgataagct tcgcgaacct cttcgaggga cataacttcg
     4261 tataatgtat gctatacgaa gttatgggtc gatggtgaga tctggactag agggtcgatg
     4321 gtgatgcttg gatcaaaata ggaagaatcc caatgataac ttcgtataaa gtatcctata
     4381 cgaagttata ctaggtaccg gatccgccac catgaccgcc gtgagcacca cagccactac
     4441 cgtgctgcag gccacacaga gcgacgtgct gcaggagatc cagtccaact tcctgctgaa
     4501 tagctccatc tgggtgaaca ttgctctggc cggagtggtc atcctgctgt ttgtggccat
     4561 ggggagggat ctggaatccc ctagagctaa gctgatctgg gtggccacaa tgctggtgcc
     4621 actggtgtct atttctagtt acgctggact ggccagtggg ctgactgtgg gcttcctgca
     4681 gatgccacct ggacacgctc tggccggaca ggaggtgctg agcccatggg gccggtatct
     4741 gacatggact ttctccactc ccatgatcct gctggctctg ggactgctgg ccgacaccga
     4801 tattgccagc ctgttcaccg ccatcacaat ggacattggc atgtgcgtga caggactggc
     4861 cgctgccctg atcactagct cccatctgct gcgctgggtg ttctacggaa tttcttgtgc
     4921 tttctttgtg gccgtgctgt atgtgctgct ggtgcagtgg ccagctgatg ctgaggctgc
     4981 tgggaccagt gaaatctttg gcactctgag gattctgacc gtggtgctgt ggctggggta
     5041 ccctatcctg ttcgctctgg gctctgaggg agtggccctg ctgagtgtgg gagtgaccag
     5101 ctggggatac tccggactgg acatcctggc taaatacgtg ttcgcctttc tgctgctgag
     5161 atgggtggct gccaatgaag gcacagtgtc tgggagtgga atgggaatcg ggtccggagg
     5221 agctgctcca gccgacgatc gaccggtagt aaaatccaga attacttctg aaggggagta
     5281 tatccctctg gatcaaatag acatcaatgt tgcacctgca ggggcagtga gcaagggcga
     5341 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     5401 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     5461 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     5521 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     5581 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     5641 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     5701 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     5761 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     5821 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     5881 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     5941 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     6001 cgccgccggg atcactctcg gcatggacga gctgtacaag ttctgctacg agaatgaagt
     6061 gtaatgagct agctagtata acttcgtata gcatacatta tacgaagtta tgggtcgatg
     6121 gtgatgcttg gcaattcggg tcgatggtga agcattggtc ttcctatttt gatataactt
     6181 cgtataggat actttatacg aagttatggg aattcaattc tttgccaaaa tgatgagaca
     6241 gcacaataac cagcacgttg cccaggagct gtaggaaaaa gaagaaggca tgaacatggt
     6301 tagcagaggc tctagagccg ccggtcacac gccagaagcc gaaccccgcc ctgccccgtc
     6361 ccccccgaag gcagccgtcc ccccgcggac agccccgagc cgctcccccc gcacaaaggg
     6421 ccctcccgga gcccctcaag gctttcacgc agccacagaa aagaaacaag ccgtcattaa
     6481 accaagcgct aattacagcc cggaggagaa gggccgtccc gcccgctcac ctgtgggagt
     6541 aacgcggtca gtcagagccg gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg
     6601 cgaaggcagc gcgcagcgac tcccgcccgc cgcgcgcttc gctttttata gggccgccgc
     6661 cgccgccgcc tcgccataaa aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc
     6721 gcacctctcc gcctcgcccc gccccgcccc tcgccccgcc ccgccccgcc tggcgcgcgc
//
