LOCUS       sequence-196698-        5146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BLInCR 'Localizer' construct that marks tetO arrays and is targeted 
            by a PHR-tagged effector upon illumination with blue light.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5146)
  AUTHORS   Rademacher A, Erdel F, Trojanowski J, Schumacher S, Rippe K
  TITLE     Real-time observation of light-controlled transcription in living 
            cells.
  JOURNAL   J Cell Sci. 2017 Dec 15;130(24):4213-4224. doi: 10.1242/jcs.205534. 
            Epub 2017 Nov 9.
  PUBMED    29122982
REFERENCE   2  (bases 1 to 5146)
  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: 
            "10.1242/jcs.205534"; journalName: "J Cell Sci"; date: "2017-12-15- 
            15"; volume: "130"; issue: "24"; pages: "4213-4224"
COMMENT     Sequence Label: pCIBN-TetR
FEATURES             Location/Qualifiers
     source          1..5146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             625..1134
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     CDS             1147..1167
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1171..1800
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MVSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSD"
     primer_bind     complement(1247..1266)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    1934..2055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1971..1990)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2025..2044
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2062..2517)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2199..2220)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2411..2430
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2544..2648
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2650..3007
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2650..2841
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2673..2693)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2858..2993
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3042..3836
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3042..3836
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3096..3115)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3706..3725
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4024..4043)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4068..4115
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4444..5032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4933..4952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatgaat ggagctatag gaggtgacct tttgctcaat
      661 tttcctgaca tgtcggtcct agagcgccaa agggctcacc tcaagtacct caatcccacc
      721 tttgattctc ctctcgccgg cttctttgcc gattcttcaa tgattaccgg cggcgagatg
      781 gacagctatc tttcgactgc cggtttgaat cttccgatga tgtacggtga gacgacggtg
      841 gaaggtgatt caagactctc aatttcgccg gaaacgacgc ttgggactgg aaatttcaag
      901 gcagcgaagt ttgatacaga gactaaggat tgtaatgagg cggcgaagaa gatgacgatg
      961 aacagagatg acctagtaga agaaggagaa gaagagaagt cgaaaataac agagcaaaac
     1021 aatgggagca caaaaagcat caagaagatg aaacacaaag ccaagaaaga agagaacaat
     1081 ttctctaatg attcatctaa agtgacgaag gaattggaga aaacggatta tattcatgcc
     1141 cgggatccaa aaaagaagag aaaggtagcc atggtgtcta gattagataa aagtaaagtg
     1201 attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa
     1261 ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg
     1321 gctttgctcg acgccttagc cattgagatg ttagataggc accatactca cttttgccct
     1381 ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct
     1441 ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag
     1501 tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat
     1561 gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa
     1621 gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta
     1681 ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc
     1741 cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtctgactaa
     1801 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     1861 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     1921 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1981 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2041 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2101 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2161 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2221 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2281 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2341 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2401 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2461 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2521 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2581 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2641 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2701 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2761 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2821 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2881 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     2941 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3001 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3061 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3121 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3181 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     3241 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3301 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3361 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3421 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3481 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3541 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3601 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3661 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3721 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3781 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3841 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3901 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3961 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4021 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4081 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4141 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4201 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4261 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4321 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4381 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4441 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4501 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4561 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     4621 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4681 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4741 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4801 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4861 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4921 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4981 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5041 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5101 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
