LOCUS       sequence-116873-        5771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSC101 based plasmid where pl-tetO expression drives synthesis of 
            construct expressing (N to C terminal) MBP (Maltose Binding 
            Protein), Ntag, and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5771)
  AUTHORS   Sekar K, Gentile AM, Bostick JW, Tyo KE
  TITLE     N-Terminal-Based Targeted, Inducible Protein Degradation in 
            Escherichia coli.
  JOURNAL   PLoS One. 2016 Feb 22;11(2):e0149746. doi: 
            10.1371/journal.pone.0149746. eCollection 2016.
  PUBMED    26900850
REFERENCE   2  (bases 1 to 5771)
  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: "PLoS One.";
            date: "2016-02-22"; volume: "11(2):e0149746. doi"; pages: " 
            10.1371/journal.pone.0149746. eCollection 2016"
COMMENT     Sequence Label: pKS011
FEATURES             Location/Qualifiers
     source          1..5771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      272..358
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      407..501
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      993..1215
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1263..2213
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(2426..2520)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2543..3202)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3117..3136
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3203..3305)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3351..3369
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    3390..3408
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        join(3415..3433,3434..3439,3440..3456,3457..3462,
                     3463..3487)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    3415..3433
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3440..3458
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             join(3497..3574,3575..4672)
                     /codon_start=1
                     /gene="malE"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This gene encodes a signal sequence that can direct 
                     MBP to the bacterial periplasm."
                     /translation="MKIKTGARILALSALTTMMFSASALAKIEEGKLVIWINGDKGYNG
                     LAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAE
                     ITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDK
                     ELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLV
                     DLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPS
                     KPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKD
                     PRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     4646..4669
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             5033..5056
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5057..5767
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5202..5220)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5470..5489
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 tatccgggta ggcgcaatca ctttcgtctc tactccgtta caaagcgagg ctgggtattt
       61 cccggccttt ctgttatccg aaatccactg aaagcacagc ggctggctga ggagataaat
      121 aataaacgag gggctgtatg cacaaagcat cttctgttga gttaagaacg agtatcgaga
      181 tggcacatag ccttgctcaa attggaatca ggtttgtgcc aataccagta gaaacagacg
      241 aaggatccca tggtacgcgt gctagaggca tcaaataaaa cgaaaggctc agtcgaaaga
      301 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
      361 gccgccctag acctaggctg tcagaccaag tttacgagct cgcttggact cctgttgata
      421 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
      481 gcgtttttta ttggtgagaa tccaagcact agggacagta agacgggtaa gcctgttgat
      541 gataccgctg ccttactggg tgcattagcc agtctgaatg acctgtcacg ggataatccg
      601 aagtggtcag actggaaaat cagagggcag gaactgctga acagcaaaaa gtcagatagc
      661 accacatagc agacccgcca taaaacgccc tgagaagccc gtgacgggct tttcttgtat
      721 tatgggtagt ttccttgcat gaatccataa aaggcgcctg tagtgccatt tacccccatt
      781 cactgccaga gccgtgagcg cagcgaactg aatgtcacga aaaagacagc gactcaggtg
      841 cctgatggtc ggagacaaaa ggaatattca gcgatttgcc cgagcttgcg agggtgctac
      901 ttaagccttt agggttttaa ggtctgtttt gtagaggagc aaacagcgtt tgcgacatcc
      961 ttttgtaata ctgcggaact gactaaagta gtgagttata cacagggctg ggatctattc
     1021 tttttatctt tttttattct ttctttattc tataaattat aaccacttga atataaacaa
     1081 aaaaaacaca caaaggtcta gcggaattta cagagggtct agcagaattt acaagttttc
     1141 cagcaaaggt ctagcagaat ttacagatac ccacaactca aaggaaaagg acatgtaatt
     1201 atcattgact agcccatctc aattggtata gtgattaaaa tcacctagac caattgagat
     1261 gtatgtctga attagttgtt ttcaaagcaa atgaactagc gattagtcgc tatgacttaa
     1321 cggagcatga aaccaagcta attttatgct gtgtggcact actcaacccc acgattgaaa
     1381 accctacaag gaaagaacgg acggtatcgt tcacttataa ccaatacgct cagatgatga
     1441 acatcagtag ggaaaatgct tatggtgtat tagctaaagc aaccagagag ctgatgacga
     1501 gaactgtgga aatcaggaat cctttggtta aaggctttga gattttccag tggacaaact
     1561 atgccaagtt ctcaagcgaa aaattagaat tagtttttag tgaagagata ttgccttatc
     1621 ttttccagtt aaaaaaattc ataaaatata atctggaaca tgttaagtct tttgaaaaca
     1681 aatactctat gaggatttat gagtggttat taaaagaact aacacaaaag aaaactcaca
     1741 aggcaaatat agagattagc cttgatgaat ttaagttcat gttaatgctt gaaaataact
     1801 accatgagtt taaaaggctt aaccaatggg ttttgaaacc aataagtaaa gatttaaaca
     1861 cttacagcaa tatgaaattg gtggttgata agcgaggccg cccgactgat acgttgattt
     1921 tccaagttga actagataga caaatggatc tcgtaaccga acttgagaac aaccagataa
     1981 aaatgaatgg tgacaaaata ccaacaacca ttacatcaga ttcctaccta cgtaacggac
     2041 taagaaaaac actacacgat gctttaactg caaaaattca gctcaccagt tttgaggcaa
     2101 aatttttgag tgacatgcaa agtaagcatg atctcaatgg ttcgttctca tggctcacgc
     2161 aaaaacaacg aaccacacta gagaacatac tggctaaata cggaaggatc tgaggttctt
     2221 atggctcttg tatctatcag tgaagcatca agactaacaa acaaaagtag aacaactgtt
     2281 caccgttaga tatcaaaggg aaaactgtcc atatgcacag atgaaaacgg tgtaaaaaag
     2341 atagatacat cagagctttt acgagttttt ggtgcattta aagctgttca ccatgaacag
     2401 atcgacaatg taacctagtg cttggattct caccaataaa aaacgcccgg cggcaaccga
     2461 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     2521 caagcgagct cccttaaaaa aattacgccc cgccctgcca ctcatcgcag tactgttgta
     2581 attcattaag cattctgccg acatggaagc catcacagac ggcatgatga acctgaatcg
     2641 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     2701 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     2761 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     2821 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     2881 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     2941 cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc ggatgagcat
     3001 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     3061 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     3121 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     3181 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     3241 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     3301 gatcaacgga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3361 tcacgaggcc ctttcgtctt cacctcgagt ccctatcagt gatagagatt gacatcccta
     3421 tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacat cagcaggacg
     3481 cactgacagg taccgcatga aaataaaaac aggtgcacgc atcctcgcat tatccgcatt
     3541 aacgacgatg atgttttccg cctcggctct cgccaaaatc gaagaaggta aactggtaat
     3601 ctggattaac ggcgataaag gctataacgg tctcgctgaa gtcggtaaga aattcgagaa
     3661 agataccgga attaaagtca ccgttgagca tccggataaa ctggaagaga aattcccaca
     3721 ggttgcggca actggcgatg gccctgacat tatcttctgg gcacacgacc gctttggtgg
     3781 ctacgctcaa tctggcctgt tggctgaaat caccccggac aaagcgttcc aggacaagct
     3841 gtatccgttt acctgggatg ccgtacgtta caacggcaag ctgattgctt acccgatcgc
     3901 tgttgaagcg ttatcgctga tttataacaa agatctgctg ccgaacccgc caaaaacctg
     3961 ggaagagatc ccggcgctgg ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt
     4021 caacctgcaa gaaccgtact tcacctggcc gctgattgct gctgacgggg gttatgcgtt
     4081 caagtatgaa aacggcaagt acgacattaa agacgtgggc gtggataacg ctggcgcgaa
     4141 agcgggtctg accttcctgg ttgacctgat taaaaacaaa cacatgaatg cagacaccga
     4201 ttactccatc gcagaagctg cctttaataa aggcgaaaca gcgatgacca tcaacggccc
     4261 gtgggcatgg tccaacatcg acaccagcaa agtgaattat ggtgtaacgg tactgccgac
     4321 cttcaagggt caaccatcca aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc
     4381 cagtccgaac aaagagctgg caaaagagtt cctcgaaaac tatctgctga ctgatgaagg
     4441 tctggaagcg gttaataaag acaaaccgct gggtgccgta gcgctgaagt cttacgagga
     4501 agagttggtg aaagatccgc gtattgccgc cactatggaa aacgcccaga aaggtgaaat
     4561 catgccgaac atcccgcaga tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa
     4621 cgccgccagc ggtcgtcaga ctgtcgatga agccctgaaa gacgcgcaga ctaattcgag
     4681 ctcgaacaac aacaacaata acaataacaa caacctcggg tcgatcacgt cgctgtataa
     4741 gaaagccggt tcggaaaacc tgtattttca atttcataaa agcggtgcct ggaagctgcc
     4801 ggtgagtctg gtgaaacgtg gtatcgataa actggattat aaagaacagc ttcaagcatg
     4861 gcgctgggaa cgcgaaattg atgaacgcaa ccgtccgctg tcagatgagg aactcgatgc
     4921 aatgtttcct gaaggttaca aggttttacc accgccggcc ggctacgtac cgatccgcac
     4981 gcccgcacac atggatcgta tcccggcagc acgcaccact gaaaacagta gcgactacaa
     5041 agatgacgat gacaagatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
     5101 gttcatgcgc ttcaaggttc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
     5161 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     5221 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     5281 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     5341 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     5401 ctcctccttg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     5461 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     5521 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     5581 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     5641 cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat
     5701 cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta
     5761 caagtaataa a
//
