LOCUS       sequence-246418-        6231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV genome with tet-inducible, Cre-dependent expression of 
            farnesylated (f) tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6231)
  AUTHORS   Rajendran PS, Challis RC, Fowlkes CC, Hanna P, Tompkins JD, Jordan 
            MC, Hiyari S, Gabris-Weber BA, Greenbaum A, Chan KY, Deverman BE, 
            Munzberg H, Ardell JL, Salama G, Gradinaru V, Shivkumar K
  TITLE     Identification of peripheral neural circuits that regulate heart 
            rate using optogenetic and viral vector strategies.
  JOURNAL   Nat Commun. 2019 Apr 26;10(1):1944. doi: 10.1038/s41467-019-09770-1.
  PUBMED    31028266
REFERENCE   2  (bases 1 to 6231)
  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: "Nat 
            Commun."; date: "2019-04-26"; volume: "10(1):1944. doi"; pages: " 
            10.1038/s41467-019-09770-1"
COMMENT     Sequence Label: pAAV-TRE-DIO-tdTomato-f
FEATURES             Location/Qualifiers
     source          1..6231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    165..183
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    202..220
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    239..257
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    276..294
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    313..331
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    350..368
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    387..405
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    424..442
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     464..488
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(587..620)
                     /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(671..704)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2200..2209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2216..2249
                     /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    2300..2333
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2358..2946
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2411..2431)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2829..2840)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2978..3454
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3177..3196)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3494..3634
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3494..3623
                     /label=AAV2 ITR
     rep_origin      3709..4164
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3796..3815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4006..4027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4181..4200)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4300..4322
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4360..4378)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4446..4550
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4551..4619,4620..5411)
                     /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(4769..4788)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5582..6170
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6071..6090
                     /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 gcggccgcac gcgtatcaag tgccacctga cgtctcccta tcagtgatag
      181 agaagtcgac acgtctcgag ctccctatca gtgatagaga aggtacgtct agaacgtctc
      241 cctatcagtg atagagaagt cgacacgtct cgagctccct atcagtgata gagaaggtac
      301 gtctagaacg tctccctatc agtgatagag aagtcgacac gtctcgagct ccctatcagt
      361 gatagagaag gtacgtctag aacgtctccc tatcagtgat agagaagtcg acacgtctcg
      421 agctccctat cagtgataga gaagctaccc cctatataag cagagctcgt ttagtgaacc
      481 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
      541 gatccagcct ggatccggta ccggatcctc tagagtcgac tccggaataa cttcgtatag
      601 gatactttat acgaagttat gcagaatggt agctggattg tagctgctat tagcaatatg
      661 aaacctctta ataacttcgt atagcataca ttatacgaag ttatggcgcg cctcaggaga
      721 gcacacactt gcagctcatg cagccggggc cactctcatc aggagggttc agctttttat
      781 acagctcatc catcccatag agaaagaggt ggtgtctacc ctcagatcgc tcatactgtt
      841 cgacaattgt gtagtcctca ttgtgtgagg ttatgtccag ctttgtgtct acgtagtagt
      901 acccgggcaa ttgcacgggc ttcttggcca tgtatatggt cttaaactcc accaggtaat
      961 gtccgccatc tttgagtttc agagcttggt ggatctcacc cttcagcact ccatccctag
     1021 gatacagtct ctcggtgctt gcttcccagc ccattgtttt cttctgcata acggggccat
     1081 cgggtggaaa gttagttcca cgcattttca ctttgtaaat cagagtccca tcctgcagac
     1141 tggaatcctg ggtcacagtt acaagcccac catcctcgaa gttcataacc cgctcccact
     1201 taaagccttc tggaaaggag agcttcttgt agtccgggat gtcagctggg tgtttgacat
     1261 aggccttgct accgtacata aactggggag acaggatgtc ccaagcaaaa ggcagaggtc
     1321 cgcccttggt taccttcagc ttggctgtct gagtgccctc gtagggtcta ccctcccctt
     1381 caccctcgat ttcgaattcg tggccattca ttgatccctc catacgaact ttgaacctca
     1441 tgaactcctt aatcacggcc atgttgttat cctcgctgga tgcagtgcca gatgatccag
     1501 atcctgtgct gcctgttcca tggccaagaa acagatggtg cctcccctca gagcgttcat
     1561 actgttccac gatggtatag tcctcgttgt gacttgtaat gtcaagttta gtgtccacat
     1621 agtagtaacc cgggagctgt actggtttct ttgccatata gatcgtctta aattcgacca
     1681 gatagtggcc gccgtccttg agtttaagtg cctgatgtat ctcgcccttc agaacaccgt
     1741 cccgtgggta caggcgttcc gtacttgctt cccaccccat ggttttcttt tgcatgacag
     1801 gaccatcagg agggaaatta gtgcccctca tcttgacctt gtagatcagt gttccgtcct
     1861 gcaagctgct atcttgggtg acggtcacga gtccgccatc ctcgaaattc atcactcttt
     1921 cccacttgaa cccctccggg aaggacagtt tcttatagtc gggaatatcg gcaggatgct
     1981 tcacataggc cttacttccg tacatgaact gaggggacag aatgtcccaa gcgaatggca
     2041 agggtcctcc tttggtcact ttcaatttcg ccgtctgtgt cccctcgtat ggtcggccct
     2101 ctccctcccc ttcgatctcg aattcatggc cgttcatgct cccttccatg cgaactttaa
     2161 accgcatgaa ttccttgatg acttcctctc cctttgacac catggtggcg ctagcataac
     2221 ttcgtataaa gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt
     2281 ctttagaatg gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcg
     2341 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     2401 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     2461 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     2521 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     2581 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     2641 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     2701 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     2761 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     2821 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     2881 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     2941 ccccgcatcg ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc
     3001 cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat
     3061 aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag
     3121 gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta
     3181 ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg
     3241 ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac
     3301 caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct
     3361 ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt
     3421 acaggcgtga accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg
     3481 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3541 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3601 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     3661 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3721 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3781 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3841 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3901 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3961 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4021 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4081 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4141 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4201 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4261 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4321 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4381 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4441 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4501 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4561 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4621 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4681 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4741 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4801 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4861 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4921 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4981 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5041 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5101 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5161 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5221 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5281 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5341 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5401 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5461 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5521 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5581 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5641 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5701 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5761 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5821 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5881 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5941 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6001 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6061 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
