LOCUS       sequence-107337-        3924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Photoactivatable transcription system. mCherry reporter under the 
            control of a tet-inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3924)
  AUTHORS   Nihongaki Y, Yamamoto S, Kawano F, Suzuki H, Sato M
  TITLE     CRISPR-Cas9-based Photoactivatable Transcription System.
  JOURNAL   Chem Biol. 2015 Feb 19;22(2):169-74. doi: 
            10.1016/j.chembiol.2014.12.011. Epub 2015 Jan 22.
  PUBMED    25619936
REFERENCE   2  (bases 1 to 3924)
  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: 
            "10.1016/j.chembiol.2014.12.011"; journalName: "Chem Biol"; date: 
            "2015-02-19- 19"; volume: "22"; issue: "2"; pages: "169-74"
COMMENT     Sequence Label: Tet-inducible mCherry reporter
FEATURES             Location/Qualifiers
     source          1..3924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(29..47,48..70,71..89,90..112,113..131,132..154,
                     155..173,174..196,197..215,216..238,239..257,258..280,
                     281..299)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(71..89,90..112,113..131,132..154,155..173,174..196,
                     197..215,216..238,239..257,258..280,281..299,300..322,
                     323..341)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(113..131,132..154,155..173,174..196,197..215,216..238,
                     239..257,258..280,281..299,300..322,323..341,342..364,
                     365..383)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(155..173,174..196,197..215,216..238,239..257,258..280,
                     281..299,300..322,323..341,342..364,365..383,384..406,
                     407..425)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(197..215,216..238,239..257,258..280,281..299,300..322,
                     323..341,342..364,365..383,384..406,407..425,426..448,
                     449..467)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(239..257,258..280,281..299,300..322,323..341,342..364,
                     365..383,384..406,407..425,426..448,449..467,468..490,
                     491..509)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(281..299,300..322,323..341,342..364,365..383,384..406,
                     407..425,426..448,449..467,468..490,491..509,510..532,
                     533..551)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     primer_bind     complement(782..801)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     997..1021
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1106..1115
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1112..1822
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1257..1275)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1525..1544
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1840..1961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1877..1896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1931..1950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1995..2012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2166..2754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2246..2265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2925..3716,3717..3785))
                     /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     3548..3567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3786..3890)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgatatcc ctcgatcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg
       61 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc
      121 agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga
      181 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
      241 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
      301 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
      361 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat
      421 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg
      481 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc
      541 agtgatagag aaaagtgaaa gtcgagctcg gtacgctatg gcatgcatgt gtcgcgacct
      601 gcaggccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      661 gaccaccctg acctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
      721 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
      781 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
      841 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
      901 ggagtacaac gccatcagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat
      961 caaggccaac ttcaagagct agccctatat aagcagagct cgtttagtga accgtcagat
     1021 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     1081 cctccgcggc cccggtaccg aattcgccac catggtgagc aagggcgagg aggataacat
     1141 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     1201 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1261 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1321 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     1381 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1441 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1501 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1561 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     1621 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     1681 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     1741 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     1801 catggacgag ctgtacaagt aaagatctta ttaaagcaga acttgtttat tgcagcttat
     1861 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1921 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gtcgactcta
     1981 gaatcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2041 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2101 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2161 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2221 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2281 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2341 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2401 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2461 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2521 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2581 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2641 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2701 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2761 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2821 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2881 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2941 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3001 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3061 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3121 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3181 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3241 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3301 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3361 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3421 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3481 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3541 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3601 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3661 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3721 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3781 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3841 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3901 cgaaaagtgc cacctgacgt cgtc
//
