LOCUS       sequence-161494-        7105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of Chronos-tdTomato under the Syn promoter, 
            in floxed/reversed (Cre-dependent) manner. Using bGHpA signal. 
            tdTomato is a codon diversified version..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7105)
  AUTHORS   Klapoetke NC, Murata Y, Kim SS, Pulver SR, Birdsey-Benson A, Cho YK,
            Morimoto TK, Chuong AS, Carpenter EJ, Tian Z, Wang J, Xie Y, Yan Z, 
            Zhang Y, Chow BY, Surek B, Melkonian M, Jayaraman V, 
            Constantine-Paton M, Wong GK, Boyden ES
  TITLE     Independent optical excitation of distinct neural populations.
  JOURNAL   Nat Methods. 2014 Mar;11(3):338-46. doi: 10.1038/nmeth.2836. Epub 
            2014 Feb 9.
  PUBMED    24509633
REFERENCE   2  (bases 1 to 7105)
  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/nmeth.2836"; journalName: "Nat Methods"; date: "2014-03"; 
            volume: "11"; issue: "3"; pages: "338-46"
COMMENT     Sequence Label: pAAV-Syn-FLEX-rc [Chronos-tdTomato]
FEATURES             Location/Qualifiers
     source          1..7105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2691..3138
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    3154..3187
                     /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    3250..3283
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      5714..5723
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5745..5778)
                     /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(5849..5882)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5915..6503
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5968..5988)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6386..6397)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6544..6751
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6921..7050
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa gtgtctagac tgcagagggc cctgcgtatg agtgcaagtg
     2701 ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca
     2761 ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg
     2821 ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct
     2881 tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca
     2941 ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc
     3001 ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg
     3061 cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg
     3121 tcgtgcctga gagcgcagtc gagaagaatt cccataactt cgtataaagt atcctatacg
     3181 aagttatatc aaaataggaa gaccaatgct tcaccatcga cccgaattgc caagcatcac
     3241 catcgaccca taacttcgta taatgtatgc tatacgaagt tatactagct agctcattac
     3301 ttatacagct catccatgcc gtacagaaac agatggtggc gtccttcaga cctctcatac
     3361 tgttccacaa tggtgtagtc ttcattgtgg gatgtgatat ccagcttggt gtccacatag
     3421 taatagccgg gcagctgcac tggtttcttg gccatgtaga tggttttgaa ctccaccagg
     3481 tagtggccgc cgtctttcag cttcagggcc tggtggattt cgcccttcag cactccgtct
     3541 ctaggataca gccgttcagt gctggcctcc caccccattg ttttcttctg catcacgggc
     3601 ccatcgggtg gaaaattggt gccgcgcatt ttgactttgt agatcagagt gccatcctgc
     3661 agagaggagt cctgtgtcac ggtcaccagg cctccatctt caaaattcat cactctctcc
     3721 cacttaaagc cctctgggaa agacagtttc ttgtaatcag gaatgtcagc gggatgcttg
     3781 acataggcct tacttccgta cataaactga ggagacagga tatcccaagc gaaaggcagt
     3841 ggtcctccct tggtcacttt cagcttggca gtctgtgtgc cctcgtaagg cctcccttcg
     3901 ccctctcctt caatctcaaa ttcgtgccca ttcatgctgc cctccatgcg caccttaaac
     3961 ctcatgaact ctttgatcac agccatattg ttatcctcgc ttgaggcggt tccactagac
     4021 cctgagccgg tacttccagt cccatggccc aggaacagat ggtgtctccc ctcgctccgt
     4081 tcatactgct ccacaatagt gtagtcctcg ttgtgtgaag tgatatccag ttttgtgtcc
     4141 acgtaatagt agcctggcag ctgcacaggt ttcttggcca tgtagattgt cttgaactcc
     4201 accaggtaat gtcccccatc tttcagcttc agagcctggt ggatttctcc tttcagcacc
     4261 ccgtcgcgtg gatacagcct ttcggtggag gcctcccatc ccatagtttt cttctgcatc
     4321 acaggtccat cagggggaaa atttgtccct ctcattttca ccttgtagat cagggtccca
     4381 tcctgcaggg agctgtcctg ggtcacagtc accaggcctc cgtcctcgaa gttcatcact
     4441 ctttcccact taaacccctc ggggaagctc agtttcttat aatcgggaat gtcagctgga
     4501 tgtttcacat aggccttgga gccgtacata aactgtgggc tcaggatgtc ccaagcgaat
     4561 ggcagaggcc cgccctttgt cactttcagc ttggctgtct gggtgccctc gtagggccgt
     4621 ccctctcctt ctccctcaat ttcgaactcg tggccgttca tgcttccttc cattctcacc
     4681 ttaaaccgca tgaactcttt gatcacttcc tcgcccttac tcactgctac taccggtgcc
     4741 gccactcctc cctcctcctc ctcgtcgacg aaggtctcga tctccatgtt ctcgccggcc
     4801 acgttgatgg tggttgtctt ccggatgtcg ccgtggatca ggatgtgctc gtggatcttc
     4861 actctcaggt ggtggcccag gaagccccaa gtgttcttgc tgatcagatc caggatcagg
     4921 tggccgattc cgctggtgta aggtgtgatc aggcccagtc cctcgtgtcc ggcgatgaac
     4981 atcacgggga acatcagcca agagcagaag aagacgtagg ccatgatctt gcaaatcttc
     5041 cggcagacgc ctttgggcag ggtgtggtag ctctcgatat acaccttggc gatctggaag
     5101 aagcaagtca cgccgtagaa caggccgatc atgaagaaca ggatcttcag ggggcccttt
     5161 gtaaaggcgg ctgtgatccc ccacacgatg ttgccgatgt cggtcaccag gatggtcatg
     5221 gtccgcttgc tgtactcttc gtgcagtccg gtcaggttgc tcaggtggat caggatcacg
     5281 gggcaagtca ggagccacat gctgtaccgc agccaaatca cggctcctcc gttggtctgg
     5341 tacactgtgg ctgggctgtc gacctcgtgg aacagctcga tgaagcactt gacgagctcg
     5401 atcacgcaga cgtagacctc ctcccagccg gttgtggcct tgaagctgtg ccaaccgtag
     5461 aagatgagct gcacgatagc cacgatcacg gtgaaccact ggagtcccac tgcgatcttg
     5521 tgctgcagct cggttccgtg gttgatgtgg gggaagcaac cgtgatcggc tccggctgtt
     5581 gtggcgttag aggtttctcc gtgagcgtct gctgctggtg tcaccactgc tgcggcgctc
     5641 agcaggcctc taattgtggc ttcggcgcta ggcacggctg agataaaggc gtgggtcatt
     5701 gtggcggctg tttccatggt ggcggcgcgg cctaggtacc tagtataact tcgtatagga
     5761 tactttatac gaagttatca ttgggattct tcctattttg atccaagcat caccatcgac
     5821 cctctagtcc agatctcacc atcgacccat aacttcgtat agcatacatt atacgaagtt
     5881 atgtccctcg aagaggttcg cgaagcttat cgataatcaa cctctggatt acaaaatttg
     5941 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     6001 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     6061 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     6121 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     6181 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     6241 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     6301 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     6361 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6421 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6481 ctccctttgg gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc tcataaagct
     6541 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     6601 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     6661 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     6721 attgggaaga caatagcagg catgctgggg atgcggtccg gactgtacaa ttgcattcat
     6781 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     6841 aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg
     6901 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     6961 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     7021 ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg
     7081 agaggcggtt tgcgtattgg gcgct
//
