LOCUS       sequence-329441-        6345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Cre-dependent SwiChR-eYFP under control of the TRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6345)
  AUTHORS   Sabatini PV, Wang J, Rupp AC, Affinati AH, Flak JN, Li C, Olson DP, 
            Myers MG Jr
  TITLE     tTARGIT AAVs mediate the sensitive and flexible manipulation of 
            intersectional neuronal populations in mice.
  JOURNAL   eLife. 2021 Mar 11;10. pii: 66835. doi: 10.7554/eLife.66835.
  PUBMED    33704065
REFERENCE   2  (bases 1 to 6345)
  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: "eLife."; 
            date: "2021-03-11"; volume: "10. pii: 66835. doi"; pages: " 
            10.7554/eLife.66835"
COMMENT     Sequence Label: pAAV-TRE-DIO-SwiChR-eYFP
FEATURES             Location/Qualifiers
     source          1..6345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(44..62,63..85,86..104,105..127,128..146,147..169,
                     170..188,189..211,212..230,231..253,254..272,273..295,
                     296..314)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        347..385
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    complement(520..553)
                     /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(564..597)
                     /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             complement(join(653..1366,1367..1369))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(698..719)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1047..1066
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1306..1327
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2362..2371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2405..2438
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2449..2482
                     /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    2506..3094
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2559..2579)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2977..2988)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3129..3146)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3135..3342
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3365..3494
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3556..4144)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3636..3655)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4315..5106,5107..5175))
                     /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     4938..4957
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5176..5280)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5348..5366
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5404..5426)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5526..5545
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5562..6017)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5699..5720)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5911..5930
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6092..6232
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6103..6232
                     /label=AAV2 ITR
ORIGIN
        1 ggttcactaa acgagctctg cttatatagg tcgagtttac cactccctat cagtgataga
       61 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt
      121 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
      181 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
      241 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct
      301 atcagtgata gagaaaagtg aaagtcgagc tcggtacccg ggtcgaggta ggcgtgtacg
      361 gtgggaggcc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
      421 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggccccga
      481 attcgagctc ggtacccggg gatcctctag tcgacgtcga taacttcgta tagcatacat
      541 tatacgaagt tatccgaatc gcaataactt cgtataggat actttatacg aagttatttg
      601 gcgcgccaaa tttaaataac gcaacgtacc ttcgaggggg cccaaggaat tcttacttgt
      661 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
      721 gatcgcgctt ctcgttgggg tctttgctca gggcggactg gtagctcagg tagtggttgt
      781 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
      841 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
      901 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
      961 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     1021 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     1081 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     1141 ggtcggggta gcgggcgaag cactgcaggc cgtagccgaa ggtggtcacg agggtgggcc
     1201 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
     1261 catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct
     1321 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc acgttgatgt
     1381 cgatctggtc cagggggatg tactcgccct cgctggtgat cctgctcttg gcggccgcga
     1441 ccgcgccagc ctcggcctcg tcctccacca gcgtctcgac ctcaatctca gtgccaccaa
     1501 tgttcaattt ggtggtcttg cgaatgtcgc cgtggatgag gatatgctcg tggatcagca
     1561 cgcgcaggta gtggccgagc agaccccagc actgcttcga catcaggtca atgatggtgt
     1621 ggccgacgtt ggagccgtac ttgctcagga cgccgaagcc ctcggggccg aggatgaaca
     1681 ggatggggaa cataccccat gatacgaaga agagccaagc catgccagtc accacctggc
     1741 gacaccggcc cttcggcacg gtgtggtacg cctcaatgta gaccttggct gcgttgaaga
     1801 atgtgtagat gccgtagcac aggcccatca ggaagaaaat gacacggacg tatcccttgg
     1861 acagcgcggc cgtggtgccc cacacgatcg tgccgatatc tgacaccagc agacccatgg
     1921 tacgcttgtt atagtcgttc gccagacccg taaggttgct cagacggata agaatgacag
     1981 gggcggtcag cagccagctc gcgtaacgaa gccacttggt cttgttgccg ttggatgagt
     2041 agatcaccgc aggttcgtca aagctatgga aatactcgat gatgaacttg atcatcgaga
     2101 tcgtggccac gtaaatctcc tcccagccgc aagtagactt ccaggtctgg tagccgtaga
     2161 acatcaggca gagcgctgaa agcgcaaaag aaatccactg cagaatgttg gcagccaact
     2221 tctcggcatt tgttccgttg gatttaagcc aagccaagca gaagcactgg ccgttgttcg
     2281 ggatgcaaat aacagaacca ttgttctcaa gagtgtatga ggtttggaag agtccgacgg
     2341 cagacaaagc gccgccatag tccatggtgg ctagctagtt ggcgcgccaa cctggccggc
     2401 cttgataact tcgtataatg tatgctatac gaagttatcc gaatcgcaat aacttcgtat
     2461 aaagtatcct atacgaagtt atcgaagatc atcaagctta tcgataatca acctctggat
     2521 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2581 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2641 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2701 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2761 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2821 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2881 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     2941 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3001 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3061 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     3121 tcgatctgcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3181 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3241 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3301 caagggggag gattgggaag acaatagcag gcatgctggg gacacgtgcg gaccgagcgg
     3361 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3421 gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc tcagtgagcg agcgagcgcg
     3481 cagctgcctg caggacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3541 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3601 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3661 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3721 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3781 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3841 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3901 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3961 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4021 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4081 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4141 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4201 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4261 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4321 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4381 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4441 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4501 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4561 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4621 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4681 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4741 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4801 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4861 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4921 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4981 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5041 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5101 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5161 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5221 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5281 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5341 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5401 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5461 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     5521 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat
     5581 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga
     5641 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc
     5701 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     5761 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     5821 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     5881 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa
     5941 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     6001 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
     6061 tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc
     6121 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     6181 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
     6241 acgcgaaaca attctgcagg aattcggggc cgcggaggct ggatcggtcc cggtgtcttc
     6301 tatggaggtc aaaacagcgt ggatggcgtc tccaggcgat ctgac
//
