LOCUS       sequence-337531-        6034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE-ChR2-YFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6034)
  AUTHORS   Hyun JH, Nagahama K, Namkung H, Mignocchi N, Roh SE, Hannan P, 
            Krussel S, Kwak C, McElroy A, Liu B, Cui M, Lee S, Lee D, Huganir 
            RL, Worley PF, Sawa A, Kwon HB
  TITLE     Tagging active neurons by soma-targeted Cal-Light.
  JOURNAL   Nat Commun. 2022 Dec 12;13(1):7692. doi: 10.1038/s41467-022-35406-y.
  PUBMED    36509775
REFERENCE   2  (bases 1 to 6034)
  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: "2022-12-12"; volume: "13(1):7692. doi"; pages: " 
            10.1038/s41467-022-35406-y"
COMMENT     Sequence Label: pAAV-TRE-ChR2-YFP
FEATURES             Location/Qualifiers
     source          1..6034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(55..73,74..96,97..115,116..138,139..157,158..180,
                     181..199,200..222,223..241,242..264,265..283,284..306,
                     307..325)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        358..396
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             517..1443
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     CDS             join(1456..1458,1459..2169)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1498..1519)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1759..1778)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2106..2127
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2206..2794
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2259..2279)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2677..2688)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2829..2846)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2835..3042
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3065..3194
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3256..3844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3336..3355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4015..4806,4807..4875))
                     /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     4638..4657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4876..4980)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5048..5066
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5104..5126)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5226..5245
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5262..5717)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5399..5420)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5611..5630
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5792..5932
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5803..5932
                     /label=AAV2 ITR
ORIGIN
        1 ggcgatctga cggttcacta aacgagctct gcttatatag gtcgagttta ccactcccta
       61 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
      121 gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
      181 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
      241 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      301 taccactccc tatcagtgat agagaaaagt gaaagtcgag ctcggtaccc gggtcgaggt
      361 aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
      421 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
      481 cgcggccccg aattcgagct cggtacccgg ggatccatgg actatggcgg cgctttgtct
      541 gccgtcggac gcgaactttt gttcgttact aatcctgtgg tggtgaacgg gtccgtcctg
      601 gtccctgagg atcaatgtta ctgtgccgga tggattgaat ctcgcggcac gaacggcgct
      661 cagaccgcgt caaatgtcct gcagtggctt gcagcaggat tcagcatttt gctgctgatg
      721 ttctatgcct accaaacctg gaaatctaca tgcggctggg aggagatcta tgtgtgcgcc
      781 attgaaatgg ttaaggtgat tctcgagttc ttttttgagt ttaagaatcc ctctatgctc
      841 taccttgcca caggacaccg ggtgcagtgg ctgcgctatg cagagtggct gctcacttgt
      901 cctgtcatcc ttatccgcct gagcaacctc accggcctga gcaacgacta cagcaggaga
      961 accatgggac tccttgtctc agacatcggg actatcgtgt ggggggctac cagcgccatg
     1021 gcaaccggct atgttaaagt catcttcttt tgtcttggat tgtgctatgg cgcgaacaca
     1081 ttttttcacg ccgccaaagc atatatcgag ggttatcata ctgtgccaaa gggtcggtgc
     1141 cgccaggtcg tgaccggcat ggcatggctg tttttcgtga gctggggtat gttcccaatt
     1201 ctcttcattt tggggcccga aggttttggc gtcctgagcg tctatggctc caccgtaggt
     1261 cacacgatta ttgatctgat gagtaaaaat tgttgggggt tgttgggaca ctacctgcgc
     1321 gtcctgatcc acgagcacat attgattcac ggagatatcc gcaaaaccac caaactgaac
     1381 atcggcggaa cggagatcga ggtcgagact ctcgtcgaag acgaagccga ggccggagcc
     1441 gtgccagcgg ccgccgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1501 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1561 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1621 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
     1681 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1741 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1801 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1861 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1921 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1981 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2041 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     2101 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2161 ctgtacaagt tctgctacga gaacgaggtg taaaagctta tcgataatca acctctggat
     2221 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2281 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2341 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2401 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2461 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2521 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2581 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     2641 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2701 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2761 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     2821 tcgatctgcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2881 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2941 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3001 caagggggag gattgggaag acaatagcag gcatgctggg gacacgtgcg gaccgagcgg
     3061 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3121 gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc tcagtgagcg agcgagcgcg
     3181 cagctgcctg caggacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4981 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5041 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     5101 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5161 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     5221 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aaaattgtaa acgttaatat
     5281 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aatagaccga
     5341 aatcggcaaa atcccttata aatcaaaaga atagcccgag atagagttga gtgttgttcc
     5401 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     5461 cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt ttttggggtc
     5521 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     5581 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctaa
     5641 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     5701 gccgctacag ggcgcgtact atggttgctt tgacgtatgc ggtgtgaaat accgcacaga
     5761 tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc tcgctcgctc
     5821 actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc ggcctcagtg
     5881 agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc ctgcggccgc
     5941 acgcgaaaca attctgcagg aattcggggc cgcggaggct ggatcggtcc cggtgtcttc
     6001 tatggaggtc aaaacagcgt ggatggcgtc tcca
//
