LOCUS       sequence-364132-        5951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Optogenetic CIBN localizer domain coupled to reverse Tet repressor; 
            binds tetO sites upon doxycycline addition; CIBN is bound by PHR 
            upon blue light exposure.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5951)
  AUTHORS   Trojanowski J, Frank L, Rademacher A, Mucke N, Grigaitis P, Rippe K
  TITLE     Transcription activation is enhanced by multivalent interactions 
            independent of phase separation.
  JOURNAL   Mol Cell. 2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017.
  PUBMED    35537448
REFERENCE   2  (bases 1 to 5951)
  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"; journalName: "Mol Cell. 
            2022 May 4. pii: S1097-2765(22)00328-8. doi: 
            10.1016/j.molcel.2022.04.017."
COMMENT     Sequence Label: CIBN-rTetR
FEATURES             Location/Qualifiers
     source          1..5951
                     /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"
     CDS             1830..1883
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1884..2483
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1884..1903)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2380..2400
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2511..2528)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2517..2724
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    2739..2860
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2776..2795)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2830..2849
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2867..3322)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3004..3025)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3216..3235
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3349..3453
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3455..3812
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3455..3646
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3478..3498)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3663..3798
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3847..4641
                     /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             3847..4641
                     /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(3901..3920)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4511..4530
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4829..4848)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4873..4920
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5249..5837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5738..5757
                     /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 ggcagtggag agggcagagg aagtctgcta acatgcggtg
     1861 acgtcgagga gaatcctggc ccaatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     1921 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     1981 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     2041 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     2101 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     2161 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     2221 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg
     2281 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     2341 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     2401 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     2461 tgacccgcaa gcccggtgcc tgagaattct aactagagct cgctgatcag cctcgactgt
     2521 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2581 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2641 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2701 agagaatagc aggcatgctg gggacaattg ttgttgttaa cttgtttatt gcagcttata
     2761 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2821 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     2881 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2941 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3001 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3061 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3121 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3181 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3241 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3301 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3361 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3421 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3481 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3541 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3601 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3661 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3721 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3781 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3841 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3901 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3961 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4021 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4081 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4141 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4201 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4261 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4321 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4381 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4441 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4501 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4561 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4621 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4681 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4741 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4801 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4861 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4921 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4981 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5041 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5101 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5161 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5221 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5281 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5341 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5401 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5461 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5521 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5581 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5641 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5701 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5761 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5821 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5881 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5941 ccgccatgca t
//
