LOCUS       sequence-199261-        5813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  'Localizer' construct that marks tetO arrays and is targeted by a 
            PHR-tagged effector upon illumination with blue light; can be 
            visualized without triggering PHR recruitment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5813)
  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 5813)
  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-tagRFP-T
FEATURES             Location/Qualifiers
     source          1..5813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             594..1103
                     /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             1116..1739
                     /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
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(1192..1211)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             1749..2462
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    2592..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2629..2648)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2683..2702
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2720..3175)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2857..2878)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3069..3088
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3202..3306
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3308..3665
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3308..3499
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3331..3351)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3516..3651
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3700..4494
                     /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             3700..4494
                     /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(3754..3773)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4364..4383
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4682..4701)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4726..4773
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5102..5690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5591..5610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgcc accatgaatg
      601 gagctatagg aggtgacctt ttgctcaatt ttcctgacat gtcggtccta gagcgccaaa
      661 gggctcacct caagtacctc aatcccacct ttgattctcc tctcgccggc ttctttgccg
      721 attcttcaat gattaccggc ggcgagatgg acagctatct ttcgactgcc ggtttgaatc
      781 ttccgatgat gtacggtgag acgacggtgg aaggtgattc aagactctca atttcgccgg
      841 aaacgacgct tgggactgga aatttcaagg cagcgaagtt tgatacagag actaaggatt
      901 gtaatgaggc ggcgaagaag atgacgatga acagagatga cctagtagaa gaaggagaag
      961 aagagaagtc gaaaataaca gagcaaaaca atgggagcac aaaaagcatc aagaagatga
     1021 aacacaaagc caagaaagaa gagaacaatt tctctaatga ttcatctaaa gtgacgaagg
     1081 aattggagaa aacggattat attcatgtac cggtaatggt gtctagatta gataaaagta
     1141 aagtgattaa cagcgcatta gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc
     1201 gtaaactcgc ccagaagcta ggtgtagagc agcctacatt gtattggcat gtaaaaaata
     1261 agcgggcttt gctcgacgcc ttagccattg agatgttaga taggcaccat actcactttt
     1321 gccctttaga aggggaaagc tggcaagatt ttttacgtaa taacgctaaa agttttagat
     1381 gtgctttact aagtcatcgc gatggagcaa aagtacattt aggtacacgg cctacagaaa
     1441 aacagtatga aactctcgaa aatcaattag cctttttatg ccaacaaggt ttttcactag
     1501 agaatgcatt atatgcactc agcgctgtgg ggcattttac tttaggttgc gtattggaag
     1561 atcaagagca tcaagtcgct aaagaagaaa gggaaacacc tactactgat agtatgccgc
     1621 cattattacg acaagctatc gaattatttg atcaccaagg tgcagagcca gccttcttat
     1681 tcggccttga attgatcata tgcggattag aaaaacaact taaatgtgaa agtgggtctg
     1741 acaagcttat ggtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt
     1801 acatggaggg caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc
     1861 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
     1921 ccttcgacat cctggctacc agcttcatgt acggcagcag aaccttcatc aaccacaccc
     1981 agggcatccc cgacttcttt aagcagtcct tccctgaggg cttcacatgg gagagagtca
     2041 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
     2101 gcctcatcta caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatgc
     2161 agaagaaaac actcggctgg gaggccaaca ccgagatgct gtaccccgct gacggcggcc
     2221 tggaaggcag aaccgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact
     2281 tcaagaccac atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact
     2341 atgtggacca cagactggaa agaatcaagg aggccgacaa agagacctac gtcgagcagc
     2401 acgaggtggc tgtggccaga tactgcgacc tccctagcaa actggggcac aaacttaatt
     2461 aaggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     2521 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     2581 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     2641 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     2701 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     2761 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     2821 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     2881 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     2941 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     3001 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     3061 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     3121 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     3181 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3241 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3301 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3361 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3421 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3481 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3541 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3601 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3661 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3721 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3781 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3841 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     3901 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3961 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     4021 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     4081 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     4141 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     4201 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     4261 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     4321 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     4381 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     4441 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     4501 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     4561 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     4621 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     4681 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     4741 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     4801 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     4861 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     4921 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     4981 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5041 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5101 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5161 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5221 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5281 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5341 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5401 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5461 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5521 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5581 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5641 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5701 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5761 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta tta
//
