LOCUS       sequence-337530-        6070 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRE-NpHR-EYFP reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6070)
  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 6070)
  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-NpHR-EYFP
FEATURES             Location/Qualifiers
     source          1..6070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(28..46,47..69,70..88,89..111,112..130,131..153,
                     154..172,173..195,196..214,215..237,238..256,257..279,
                     280..298)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        331..369
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(1423..1425,1426..2136)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1465..1486)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1726..1745)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2073..2094
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2173..2761
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2226..2246)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2644..2655)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2796..2813)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2802..3009
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3032..3161
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3223..3811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3303..3322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3982..4773,4774..4842))
                     /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     4605..4624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4843..4947)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5015..5033
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5071..5093)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5193..5212
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5229..5684)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5366..5387)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5578..5597
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5759..5899
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5770..5899
                     /label=AAV2 ITR
     protein_bind    join(6056..6070,1..4,5..27,28..46,47..69,70..88,89..111,
                     112..130,131..153,154..172,173..195,196..214,215..237,
                     238..256)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
ORIGIN
        1 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
       61 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
      121 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
      181 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
      241 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      301 agtgaaagtc gagctcggta cccgggtcga ggtaggcgtg tacggtggga ggcctatata
      361 agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac
      421 ctccatagaa gacaccggga ccgatccagc ctccgcggcc ccgaattcga gctcggtacc
      481 cggggatcca tgaggggtac gcccctgctc ctcgtcgtct ctctgttctc tctgcttcag
      541 gacacagaga ccctgcctcc cgtgaccgag agtgccgtgg cccttcaagc cgaggttacc
      601 caaagggagt tgttcgagtt cgtgctgaac gaccctttgc ttgcaagcag tctctatatc
      661 aacatcgcac ttgcaggact gagtatactg ctgttcgttt ttatgacccg aggactcgat
      721 gatccacggg caaaacttat tgctgtgtca accatccttg tgcctgtcgt cagcattgcc
      781 tcctacactg gattggcgag cggcctgaca atttccgttc ttgaaatgcc agcgggccat
      841 tttgcagaag gcagctcagt gatgctggga ggagaagagg tagatggtgt agtcaccatg
      901 tggggacggt atctcacctg ggcactttcc acgcccatga ttctcctcgc tctgggtctc
      961 ctggccggaa gcaatgctac aaagctcttc acagctatca ctttcgatat cgctatgtgc
     1021 gtgactggcc ttgccgcggc cctgactacc tcctcccacc tcatgagatg gttctggtac
     1081 gctatcagtt gtgcatgctt tctggtggtc ttgtatatcc tgctggtgga gtgggcacag
     1141 gacgccaaag ccgcgggaac cgctgacatg ttcaataccc tgaagctgtt gacagtagtg
     1201 atgtggctgg ggtatccaat tgtgtgggct cttggagtcg agggtatcgc ggtgttgccc
     1261 gttggggtga cgagctgggg atattctttc ctggatatcg tggcaaagta cattttcgca
     1321 ttcttgctcc tgaactatct gacgtcaaac gaatctgtcg tgtccggcag cattttggat
     1381 gttccatctg cttctgggac cccggctgat gatgcggccg ccgtgagcaa gggcgaggag
     1441 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1501 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1561 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac
     1621 ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1681 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1741 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1801 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1861 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1921 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1981 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc
     2041 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2101 gccgggatca ctctcggcat ggacgagctg tacaagttct gctacgagaa cgaggtgtaa
     2161 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2221 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     2281 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     2341 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     2401 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     2461 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     2521 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     2581 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     2641 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     2701 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     2761 catcgatacc gagcgctgct cgagagatcg atctgcctcg actgtgcctt ctagttgcca
     2821 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2881 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2941 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3001 tgctggggac acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     3061 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3121 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gacatgtgag caaaaggcca
     3181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3661 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     4201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     4981 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     5041 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5101 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5161 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     5221 gcaccataaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     5281 cagctcattt tttaaccaat agaccgaaat cggcaaaatc ccttataaat caaaagaata
     5341 gcccgagata gagttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     5401 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     5461 atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     5521 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     5581 gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt gtagcggtca cgctgcgcgt
     5641 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtactatg gttgctttga
     5701 cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgcccc
     5761 tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg
     5821 gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac
     5881 tccatcacta ggggttcctg cggccgcacg cgaaacaatt ctgcaggaat tcggggccgc
     5941 ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc
     6001 aggcgatctg acggttcact aaacgagctc tgcttatata ggtcgagttt accactccct
     6061 atcagtgata
//
