LOCUS       sequence-364140-        5653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lac repressor labeled with GFP; can be used to label lacO sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5653)
  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 5653)
  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: GFP-LacI
FEATURES             Location/Qualifiers
     source          1..5653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        148..339
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     promoter        complement(341..445)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      472..927
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(559..578)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     769..790
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    934..1055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(945..964)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     999..1018
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1288..1308)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     2350..2369
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             complement(join(2407..3117,3118..3120,3121..3123))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2407..3117,3118..3120,3121..3123))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2449..2470)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2798..2817
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3057..3078
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3120..3129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3148..3477)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3162..3297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(3610..4198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3690..3709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4527..4574
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4599..4618
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4806..5600)
                     /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             complement(4806..5600)
                     /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(4917..4936)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5527..5546
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5635..5653,1..339))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5649..5653,1..131)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
       61 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
      121 aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg
      181 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa
      241 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct
      301 aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt tttcaatatt
      361 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
      421 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
      481 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
      541 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
      601 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
      661 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
      721 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
      781 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
      841 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
      901 ttaacaaaat attaacgctt acaatttacg ccttaagata cattgatgag tttggacaaa
      961 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     1021 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1081 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1141 gtggtatggc tgattatgat cagttatcta gatccggtgg atcccgggcc cgcggtaccg
     1201 tcgactgcag aattggtcgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1261 taacttacat taattgcgtt gcgctcaaac cttcctcttc ttcttaggag gcctcaggct
     1321 gcttcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1381 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     1441 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     1501 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     1561 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     1621 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     1681 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     1741 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     1801 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     1861 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     1921 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     1981 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     2041 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     2101 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     2161 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     2221 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     2281 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     2341 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacca tggatctgag
     2401 tccggacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     2461 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag
     2521 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     2581 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     2641 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     2701 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     2761 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     2821 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     2881 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     2941 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     3001 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     3061 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     3121 catggtggcg accggtagcg ctagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     3181 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     3241 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     3301 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     3361 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     3421 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     3481 ccacagtaat aactaatgca tggcggtaat acggttatcc acagaatcag gggataacgc
     3541 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3601 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3661 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3721 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3781 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3841 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3901 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3961 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4021 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4081 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4141 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4201 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4261 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4321 aagttttaaa tcaatctaaa gtatatatga gtaacctgag gctatggcag ggcctgccgc
     4381 cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa
     4441 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca
     4501 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acaccgtgcg ttttattctg
     4561 tctttttatt gccgtcatag cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt
     4621 tagcctcccc ctagggtggg cgaagaactc cagcatgaga tccccgcgct ggaggatcat
     4681 ccagccggcg tcccggaaaa cgattccgaa gcccaacctt tcatagaagg cggcggtgga
     4741 atcgaaatct cgtgatggca ggttgggcgt cgcttggtcg gtcatttcga accccagagt
     4801 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     4861 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     4921 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     4981 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     5041 gtcacgacga gatcctcgcc gtcgggcatg ctcgccttga gcctggcgaa cagttcggct
     5101 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     5161 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     5221 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     5281 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     5341 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     5401 agccgcgctg cctcgtcttg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     5461 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     5521 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     5581 aatccatctt gttcaatcat gcgaaacgat cctcatcctg tctcttgatc gatctttgca
     5641 aaagcctagg cct
//
