LOCUS       sequence-199187-        5851 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 5851)
  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 5851)
  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-tagBFP-TetR
FEATURES             Location/Qualifiers
     source          1..5851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        64..367
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        368..571
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     522..542
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             610..1119
                     /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             1138..1836
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             1882..2511
                     /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(1958..1977)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    2642..2763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2679..2698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2733..2752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2770..3225)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2907..2928)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3119..3138
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3252..3356
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3358..3715
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3358..3549
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3381..3401)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3566..3701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3750..4544
                     /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             3750..4544
                     /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(3804..3823)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4414..4433
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4732..4751)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4776..4823
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5152..5740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5641..5660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
       61 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      121 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      181 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      241 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      301 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      361 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      421 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      481 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      541 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      601 ctcgccacca tgaatggagc tataggaggt gaccttttgc tcaattttcc tgacatgtcg
      661 gtcctagagc gccaaagggc tcacctcaag tacctcaatc ccacctttga ttctcctctc
      721 gccggcttct ttgccgattc ttcaatgatt accggcggcg agatggacag ctatctttcg
      781 actgccggtt tgaatcttcc gatgatgtac ggtgagacga cggtggaagg tgattcaaga
      841 ctctcaattt cgccggaaac gacgcttggg actggaaatt tcaaggcagc gaagtttgat
      901 acagagacta aggattgtaa tgaggcggcg aagaagatga cgatgaacag agatgaccta
      961 gtagaagaag gagaagaaga gaagtcgaaa ataacagagc aaaacaatgg gagcacaaaa
     1021 agcatcaaga agatgaaaca caaagccaag aaagaagaga acaatttctc taatgattca
     1081 tctaaagtga cgaaggaatt ggagaaaacg gattatattc atggaccggt cgccaccatg
     1141 agcgagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac
     1201 catcacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg
     1261 agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctactagc
     1321 ttcctctacg gcagcaagac cttcatcaac cacacccagg gcatccccga cttcttcaag
     1381 cagtccttcc ctgagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg
     1441 ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc
     1501 agaggggtga acttcacatc caacggccct gtgatgcaga agaaaacact cggctgggag
     1561 gccttcaccg agacgctgta ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc
     1621 ctgaagctcg tgggcgggag ccatctgatc gcaaacatca agaccacata tagatccaag
     1681 aaacccgcta agaacctcaa gatgcctggc gtctactatg tggactacag actggaaaga
     1741 atcaaggagg ccaacaacga gacctacgtc gagcagcacg aggtggcagt ggccagatac
     1801 tgcgacctcc ctagcaaact ggggcacaag cttaattccg gactcagatc tcgagctcaa
     1861 gcttcgaatt ctgcagtcga catggtgtct agattagata aaagtaaagt gattaacagc
     1921 gcattagagc tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag
     1981 aagctaggtg tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc
     2041 gacgccttag ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg
     2101 gaaagctggc aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt
     2161 catcgcgatg gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact
     2221 ctcgaaaatc aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat
     2281 gcactcagcg ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa
     2341 gtcgctaaag aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa
     2401 gctatcgaat tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg
     2461 atcatatgcg gattagaaaa acaacttaaa tgtgaaagtg ggtctgacta ggggatccac
     2521 cggatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt
     2581 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     2641 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2701 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2761 ttaaggcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     2821 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     2881 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     2941 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3001 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3061 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3121 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3181 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc
     3241 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3301 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg
     3361 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3421 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3481 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3541 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3601 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3661 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg
     3721 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3781 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3841 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3901 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     3961 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4021 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4081 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4141 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4201 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4261 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4321 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4381 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4441 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4501 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4561 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4621 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4681 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg
     4741 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac
     4801 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4861 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4921 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4981 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt
     5041 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5101 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5281 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5341 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5461 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     5521 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     5581 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5641 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5701 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5761 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5821 tgattctgtg gataaccgta ttaccgccat g
//
