LOCUS       sequence-198432-        4848 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Photoactivatable transcription system. Luciferase 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 4848)
  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 4848)
  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 Luciferase reporter
FEATURES             Location/Qualifiers
     source          1..4848
                     /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"
     polyA_signal    2764..2885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2801..2820)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2855..2874
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2919..2936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3090..3678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3170..3189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3849..4640,4641..4709))
                     /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     4472..4491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4710..4814)
                     /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 catggaagat gccaaaaaca ttaagaaggg
     1141 cccagcgcca ttctacccac tcgaagacgg gaccgccggc gagcagctgc acaaagccat
     1201 gaagcgctac gccctggtgc ccggcaccat cgcctttacc gacgcacata tcgaggtgga
     1261 cattacctac gccgagtact tcgagatgag cgttcggctg gcagaagcta tgaagcgcta
     1321 tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag aatagcttgc agttcttcat
     1381 gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc ccagctaacg acatctacaa
     1441 cgagcgcgag ctgctgaaca gcatgggcat cagccagccc accgtcgtat tcgtgagcaa
     1501 gaaagggctg caaaagatcc tcaacgtgca aaagaagcta ccgatcatac aaaagatcat
     1561 catcatggat agcaagaccg actaccaggg cttccaaagc atgtacacct tcgtgacttc
     1621 ccatttgcca cccggcttca acgagtacga cttcgtgccc gagagcttcg accgggacaa
     1681 aaccatcgcc ctgatcatga acagtagtgg cagtaccgga ttgcccaagg gcgtagccct
     1741 accgcaccgc accgcttgtg tccgattcag tcatgcccgc gaccccatct tcggcaacca
     1801 gatcatcccc gacaccgcta tcctcagcgt ggtgccattt caccacggct tcggcatgtt
     1861 caccacgctg ggctacttga tctgcggctt tcgggtcgtg ctcatgtacc gcttcgagga
     1921 ggagctattc ttgcgcagct tgcaagacta taagattcaa tctgccctgc tggtgcccac
     1981 actatttagc ttcttcgcta agagcactct catcgacaag tacgacctaa gcaacttgca
     2041 cgagatcgcc agcggcgggg cgccgctcag caaggaggta ggtgaggccg tggccaaacg
     2101 cttccaccta ccaggcatcc gccagggcta cggcctgaca gaaacaacca gcgccattct
     2161 gatcaccccc gaaggggacg acaagcctgg cgcagtaggc aaggtggtgc ccttcttcga
     2221 ggctaaggtg gtggacttgg acaccggtaa gacactgggt gtgaaccagc gcggcgagct
     2281 gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac aaccccgagg ctacaaacgc
     2341 tctcatcgac aaggacggct ggctgcacag cggcgacatc gcctactggg acgaggacga
     2401 gcacttcttc atcgtggacc ggctgaagag cctgatcaaa tacaagggct accaggtagc
     2461 cccagccgaa ctggagagca tcctgctgca acaccccaac atcttcgacg ccggggtcgc
     2521 cggcctgccc gacgacgatg ccggcgagct gcccgccgca gtcgtcgtgc tggaacacgg
     2581 taaaaccatg accgagaagg agatcgtgga ctatgtggcc agccaggtta caaccgccaa
     2641 gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct aaaggactga ccggcaagtt
     2701 ggacgcccgc aagatccgcg agattctcat taaggccaag aagtaaagat cttattaaag
     2761 cagaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2821 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2881 tcttatcatg tctggtcgac tctagactct tccgcttcct cgctcactga ctcgctgcgc
     2941 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3001 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3061 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3121 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3181 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3241 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3301 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3361 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3421 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3481 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3541 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3601 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3661 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3721 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3781 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3841 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3901 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3961 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4021 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4081 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4141 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4201 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4261 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4321 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4381 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4441 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4501 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4561 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4621 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4681 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4741 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4801 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgtc
//
