LOCUS       sequence-182795-        7315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PI(WT): Photo inhibitable (blue light sensitive).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7315)
  AUTHORS   Dagliyan O, Tarnawski M, Chu PH, Shirvanyants D, Schlichting I, 
            Dokholyan NV, Hahn KM
  TITLE     Engineering extrinsic disorder to control protein activity in living
            cells.
  JOURNAL   Science. 2016 Dec 16;354(6318):1441-1444.
  PUBMED    27980211
REFERENCE   2  (bases 1 to 7315)
  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: "Science."; 
            date: "2016-12-16"; volume: "354(6318)"; pages: "1441-1444"
COMMENT     Sequence Label: pBabe-tetoff-mVenus-PICdc42
FEATURES             Location/Qualifiers
     source          1..7315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    join(1399..1417,1418..1440,1441..1459,1460..1482,1483..1501,
                     1502..1524,1525..1543,1544..1566,1567..1585,1586..1608,
                     1609..1627,1628..1650,1651..1669)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1702..1740
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1737..1761
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1856..1865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1862..1864,1865..1867,1868..2578)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1907..1928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2168..2187)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2515..2536
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2774..3202
                     /codon_start=1
                     /gene="NPH1 from Avena sativa"
                     /product="light, oxygen, or voltage domain from phototropin
                     1 of Avena sativa (Herrou and Crosson, 2011)"
                     /label=LOV2
                     /note="photoswitch that undergoes a light-inducible 
                     conformational change"
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     CDS             3683..3715
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     protein_bind    3729..3745
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        3778..4277
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3813..3836)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4210..4229
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4300..4899
                     /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(4300..4319)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4796..4816
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5408..5425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5579..6167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5659..5678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6327..7118,7119..7187))
                     /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     6950..6969
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7188..7292)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccac gaggcccttt
     1381 cgtcgtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac
     1441 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
     1501 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
     1561 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
     1621 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     1681 cgagctcggt acccgggtcg aggtaggcgt gtacggtggg aggcctatat aagcagagct
     1741 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
     1801 agacaccggg accgatccag cctccgcggc cccagatccg taaagtgaat tcgccgccac
     1861 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1921 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1981 cggcaagctg accctgaagt tgatctgcac caccggcaag ctgcccgtgc cctggcccac
     2041 cctcgtgacc accctcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
     2101 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2161 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2221 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2281 caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa
     2341 cggcatcaag gcaaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     2401 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2461 ctacctgagc taccagtcca aactgagcaa agaccccaac gagaagcgcg atcacatggt
     2521 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     2581 ttctggatcc ggtggttctg gttctggtgg taccggcgcg ccgatgcaga caattaagtg
     2641 tgttgttgtg ggcgatggtg ctgttggtaa aacatgtctc ctgatatcct acacaacaaa
     2701 caaatttcca tcggaatatg taccgactgt ttttgacaac tatgcagtca cagttatgat
     2761 tggtggatca ggtttggcta ctacacttga acgtattgag aagaactttg tcattactga
     2821 cccaagattg ccagataatc ccattatatt cgcgtccgat agtttcttgc agttgacaga
     2881 atatagccgt gaagaaattt tgggaagaaa ctgcaggttt ctacaaggtc ctgaaactga
     2941 tcgcgcgaca gtgagaaaaa ttagagatgc catagataac caaacagagg tcactgttca
     3001 gctgattaat tatacaaaga gtggtaaaaa gttctggaac ctctttcact tgcagcctat
     3061 gcgagatcag aagggagatg tccagtactt tattggggtt cagttggatg gaactgagca
     3121 tgtccgagat gctgccgaga gagagggagt catgctgatt aagaaaactg cagaaaatat
     3181 tgatgaggcg gcaaaagaac ttggctctgg tggagaacca tatactcttg gactttttga
     3241 tactgcaggg ctagaggatt atgacagatt acgaccgctg agttatccac aaacagatgt
     3301 atttctagtc tgtttttcag tggtctctcc atcttcattt gaaaacgtga aagaaaagtg
     3361 ggtgcctgag ataactcacc actgtccaaa gactcctttc ttgcttgttg ggactcaaat
     3421 tgatctcaga gatgacccct ctactattga gaaacttgcc aagaacaaac agaagcctat
     3481 cactccagag actgctgaaa agctggcccg tgacctgaag gctgtcaagt atgtggagtg
     3541 ttctgcactt acacagaaag gcctaaagaa tgtatttgac gaagcaatat tggctgccct
     3601 ggagcctcca gaaccgaaga agagccgcag gtgtgtgctg ctataaaagc ttgcggccgc
     3661 acagctgtat acacgtgcaa gccagccaga actcgccccg gaagaccccg aggatctcga
     3721 gtgtggaatt gtgagcggat aacaatttgt cgagggcccc tgcaggtcaa ttctaccggg
     3781 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
     3841 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     3901 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     3961 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     4021 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     4081 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     4141 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     4201 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     4261 catctccggg cctttcgacc tgcagccaat aagcttacca tgaccgagta caagcccacg
     4321 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     4381 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     4441 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4501 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4561 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4621 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4681 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4741 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     4801 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     4861 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
     4921 cgcccgaccg aaaggagcgc acgaccccat gcatcgataa aataaaagat tttatttagt
     4981 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     5041 acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg
     5101 taggagatcc ctgagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     5161 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     5221 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca
     5281 tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat
     5341 agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5401 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5461 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5521 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5581 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5641 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5701 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5761 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5821 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5881 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5941 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6001 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6061 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6121 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6181 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6241 tgagattatc aaaaaggatc ttcacctaga tccttttgcg gccgcaaatc aatctaaagt
     6301 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6361 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6421 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6481 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6541 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6601 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6661 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6721 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6781 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6841 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6901 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6961 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7021 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7081 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7141 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7201 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7261 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
