LOCUS       sequence-364165-        6670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lac repressor labeled with tdTomato; can be used to label lacO 
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6670)
  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 6670)
  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: tdTomato-LacI
FEATURES             Location/Qualifiers
     source          1..6670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    598..678
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      680..689
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             686..2113
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(2151..2170)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3212..3232
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3465..3586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3502..3521)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3556..3575
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3593..4048)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3730..3751)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3942..3961
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4075..4179
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4181..4538
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4181..4372
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4204..4224)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4389..4524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4573..5367
                     /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             4573..5367
                     /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(4627..4646)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5237..5256
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5555..5574)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5599..5646
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5975..6563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6464..6483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgaca
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttcgaatt ctgcagtcga cggtaccgcg
      661 ggcccgggat ccaccggtcg ccaccatggt gagcaagggc gaggaggtca tcaaagagtt
      721 catgcgcttc aaggtgcgca tggagggctc catgaacggc cacgagttcg agatcgaggg
      781 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      841 cggccccctg cccttcgcct gggacatcct gtccccccag ttcatgtacg gctccaaggc
      901 gtacgtgaag caccccgccg acatccccga ttacaagaag ctgtccttcc ccgagggctt
      961 caagtgggag cgcgtgatga acttcgagga cggcggtctg gtgaccgtga cccaggactc
     1021 ctccctgcag gacggcacgc tgatctacaa ggtgaagatg cgcggcacca acttcccccc
     1081 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcgcctgta
     1141 cccccgcgac ggcgtgctga agggcgagat ccaccaggcc ctgaagctga aggacggcgg
     1201 ccactacctg gtggagttca agaccatcta catggccaag aagcccgtgc aactgcccgg
     1261 ctactactac gtggacacca agctggacat cacctcccac aacgaggact acaccatcgt
     1321 ggaacagtac gagcgctccg agggccgcca ccacctgttc ctggggcatg gcaccggcag
     1381 caccggcagc ggcagctccg gcaccgcctc ctccgaggac aacaacatgg ccgtcatcaa
     1441 agagttcatg cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat
     1501 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     1561 caagggcggc cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc
     1621 caaggcgtac gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga
     1681 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca
     1741 ggactcctcc ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt
     1801 cccccccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg
     1861 cctgtacccc cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga
     1921 cggcggccac tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact
     1981 gcccggctac tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac
     2041 catcgtggaa cagtacgagc gctccgaggg ccgccaccac ctgttcctgt acggcatgga
     2101 cgagctgtac aagtccggac tcagatccat ggtgaaacca gtaacgttat acgatgtcgc
     2161 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     2221 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     2281 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     2341 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     2401 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     2461 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     2521 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     2581 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     2641 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     2701 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     2761 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     2821 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     2881 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     2941 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     3001 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     3061 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     3121 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3181 acgacaggtt tcccgaagca gcctgaggcc tcctaagaag aagaggaagg tttgagcgca
     3241 acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     3301 cgaccaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     3361 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     3421 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     3481 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     3541 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     3601 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3661 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3721 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3781 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3841 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3901 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3961 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4021 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4081 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4141 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     4201 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4261 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4321 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4381 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4441 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     4501 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     4561 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4621 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4681 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4741 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4801 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4861 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4921 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4981 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5041 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5101 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5161 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5221 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5281 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5341 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     5401 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5461 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5521 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     5581 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     5641 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5701 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5761 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     5821 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5881 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5941 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6001 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6061 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6121 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6181 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6241 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6301 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6361 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6421 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6481 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6541 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6601 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6661 gtattaccgc
//
