LOCUS       40924_26919        6986 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pKNG101, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6986)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6986)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 6986)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6986)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        456..484
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             696..1499
                     /codon_start=1
                     /note="unnamed protein product; StrA; streptomycin
                     resistance"
                     /protein_id="SJL87110.1"
                     /translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
                     GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
                     MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
                     ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
                     NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
     CDS             1499..2335
                     /codon_start=1
                     /note="unnamed protein product; StrB; streptomycin
                     resistance"
                     /protein_id="SJL87111.1"
                     /translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
                     IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
                     AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
                     SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
                     QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
     rep_origin      2371..2759
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     oriT            3479..3588
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             4001..4504
                     /codon_start=1
                     /note="unnamed protein product; insB"
                     /protein_id="SJL87112.1"
                     /translation="MPGNSPHYGRWPQHDFTSLKKLRPQSVTSRIQPGSDVIVCAEMDE
                     QWGYVGAKSRQRWLFYAYDSLRKTVVAHVFGERTMATLGRLMSLLSPFDVVIWMTDGWP
                     LYESRLKGKLHVISKRYTQRIERHNLNLRQHLARLGRKSLSFSKSVELHDKVIGHYLNI
                     KHYQ"
     CDS             complement(4645..6063)
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(6064..6509)
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     misc_feature    6570..6969
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
ORIGIN
        1 gtcgacggat ccccgggtac cataagtaga agcagcaacc caagtagctt taccagcatc
       61 cggttcaacc agcatagtaa gaatcttact ggacatcggc agttcttcga acagtgcgcc
      121 aactaccagc tctttctgca gttcattcag ggcaccggag aacctgcgtg caatccatct
      181 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat ccatggatta cgcgttaacc
      241 cgggcccgcg gatgcatatg atcttaaggc ctaggtctag agtctttgtt ttgacgccat
      301 tagcgtacgt aacaatcctc gttaaaggac aaggacctga gcggaagtgt atcgtacagt
      361 agacggagta tactagtata gtctatagtc cgtggaatta ttatatttat ctccgacgat
      421 attctcatca gtgaaatcca ggggaattct catgtttgac agcttatcat cgataagctt
      481 taatgcggta gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct
      541 aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg
      601 gttatgccgg tactgccggg cctcttgcgg gataactgca cattcgggat atttctctat
      661 attcgcgctt catcagaaaa ctgaaggaac ctccattgaa tcgaactaat attttttttg
      721 gtgaatcgca ttctgactgg ttgcctgtca gaggcggaga atctggtgat tttgtttttc
      781 gacgtggtga cgggcatgcc ttcgcgaaaa tcgcacctgc ttcccgccgc ggtgagctcg
      841 ctggagagcg tgaccgcctc atttggctca aaggtcgagg tgtggcttgc cccgaggtca
      901 tcaactggca ggaggaacag gagggtgcat gcttggtgat aacggcaatt ccgggagtac
      961 cggcggctga tctgtctgga gcggatttgc tcaaagcgtg gccgtcaatg gggcagcaac
     1021 ttggcgctgt tcacagccta tcggttgatc aatgtccgtt tgagcgcagg ctgtcgcgaa
     1081 tgttcggacg cgccgttgat gtggtgtccc gcaatgccgt caatcccgac ttcttaccgg
     1141 acgaggacaa gagtacgccg ctgcacgatc ttttggctcg tgtcgaacga gagctaccgg
     1201 tgcggctcga ccaagagcgc accgatatgg ttgtttgcca tggtgatccc tgcatgccga
     1261 acttcatggt ggaccctaaa actcttcaat gcacgggtct gatcgacctt gggcggctcg
     1321 gaacagcaga tcgctatgcc gatttggcac tcatgattgc taacgccgaa gagaactggg
     1381 cagcgccaga tgaagcagag cgcgccttcg ctgtcctatt caatgtattg gggatcgaag
     1441 cccccgaccg cgaacgcctt gccttctatc tgcgattgga ccctctgact tggggttgat
     1501 gttcatgccg cctgtttttc ctgctcattg gcacgtttcg caacctgttc tcattgcgga
     1561 caccttttcc agcctcgttt ggaaagtttc attgccagac gggactcctg caatcgtcaa
     1621 gggattgaaa cctatagaag acattgctga tgaactgcgc ggggccgact atctggtatg
     1681 gcgcaatggg aggggagcag tccggttgct cggtcgtgag aacaatctga tgttgctcga
     1741 atatgccggg gagcgaatgc tctctcacat cgttgccgag cacggcgact accaggcgac
     1801 cgaaattgca gcggaactaa tggcgaagct gtatgccgca tctgaggaac ccctgccttc
     1861 tgcccttctc ccgatccggg atcgctttgc agctttgttt cagcgggcgc gcgatgatca
     1921 aaacgcaggt tgtcaaactg actacgtcca cgcggcgatt atagccgatc aaatgatgag
     1981 caatgcctcg gaactgcgtg ggctacatgg cgatctgcat catgaaaaca tcatgttctc
     2041 cagtcgcggc tggctggtga tagatcccgt cggtctggtc ggtgaagtgg gctttggcgc
     2101 cgccaatatg ttctacgatc cggctgacag agacgacctt tgtctcgatc ctagacgcat
     2161 tgcacagatg gcggacgcat tctctcgtgc gctggacgtc gatccgcgtc gcctgctcga
     2221 ccaggcgtac gcttatgggt gcctttccgc agcttggaac gcggatggag aagaggagca
     2281 acgcgatcta gctatcgcgg ccgcgatcaa gcaggtgcga cagacgtcat actagatatc
     2341 tagatccggc ccgatgcgtc cggcgtagag gatctgaaga tcagcagttc aacctgttga
     2401 tagtacgtac taagctctca tgtttcacgt actaagctct catgtttaac gtactaagct
     2461 ctcatgttta acgaactaaa ccctcatggc taacgtacta agctctcatg gctaacgtac
     2521 taagctctca tgtttcacgt actaagctct catgtttgaa caataaaatt aatataaatc
     2581 agcaacttaa atagcctcta aggttttaag ttttataaga aaaaaaagaa tatataaggc
     2641 ttttaaagct tttaaggttt aacggttgtg gacaacaagc cagggatgta acgcactgag
     2701 aagcccttag agcctctcaa agcaattttc agtgacacag gaacacttaa cggctgacat
     2761 gggaattcta ctgtttgggt gtatgagcta tttgcaaagg aaattggtga tgacaaagct
     2821 cggcgctatt tgaagaaaat cgactatggc aacgccgatc ctttttgtcc ggtgttgggt
     2881 tgaaggtgaa gccggtcggg gccgcagcgg gggccggctt ttcagccttg cccccctgct
     2941 tcggccgccg tggctccggc gtcttgggtg ccggcgcggg ttccgcagcc ttggcctgcg
     3001 gtgcgggcac atcggcgggc ttggccttga tgtgccgcct ggcgtgcgag cggaacgtct
     3061 cgtaggagaa cttgaccttc cccgtttccc gcatgtgctc ccaaatggtg acgagcgcat
     3121 agccggacgc taacgccgcc tcgacatccg ccctcaccgc caggaacgca accgcagcct
     3181 catcacgccg gcgcttcttg gccgcgcggg attcaaccca ctcggccagc tcgtcggtgt
     3241 agctctttgg catcgtctct cgcctgtccc ctcagttcag taatttcctg catttgcctg
     3301 tttccagtcg gtagatattc cacaaaacag cagggaagca gcgcttttcc gctgcataac
     3361 cctgcttcgg ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata
     3421 caggattttg ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg
     3481 gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt
     3541 cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg ctggccggct accgccggcg
     3601 taacagatga gggcaagcgg atggctgatg aaaccaagcc aaccaggaag ggcagcccac
     3661 ctatcaaggt gtactgcctt ccagacgaac gaagagcgat tgaggaaaag gcggcggcgg
     3721 ccggcatgag cctgtcggcc tacctgctgg ccgtcggcca gggctacaaa atcacgggcg
     3781 tcgtggacta tgagcacgtc cgcgagctgg cccgcatcaa tggcgacctg ggccgcctgg
     3841 gcggcctgct gaaactctgg ctcaccgacg acccgcgcac ggcgcggttc ggtgatgcca
     3901 cgatccgggc aacgttgttg ccattgctgc agttcactta caccgcttct caacccggta
     3961 cgcaccataa aatcattgat atggccatga atggcgttgg atgccgggca acagcccgca
     4021 ttatgggcgt tggcctcaac acgattttac gtcacttaaa aaactcaggc cgcagtcggt
     4081 aacctcgcgc atacagccgg gcagtgacgt catcgtctgc gcggaaatgg acgaacagtg
     4141 gggctatgtc ggggctaaat cgcgccagcg ctggctgttt tacgcgtatg acagtctccg
     4201 gaagacggtt gttgcgcacg tattcggtga acgcactatg gcgacgctgg ggcgtcttat
     4261 gagcctgctg tcaccctttg acgtggtgat atggatgacg gatggctggc cgctgtatga
     4321 atcccgcctg aagggaaagc tgcacgtaat cagcaagcga tatacgcagc gaattgagcg
     4381 gcataacctg aatctgaggc agcacctggc acggctggga cggaagtcgc tgtcgttctc
     4441 aaaatcggtg gagctgcatg acaaagtcat cgggcattat ctgaacataa aacactatca
     4501 ataagttgga gtcattacca aaaggttagg aatacggtta gccatttgcc tgcttttata
     4561 tagttcatat gggattcacc tttatgttga taagaaataa aagaaaatgc caataggata
     4621 tcggcatttt cttttgcgtt tttatttgtt aactgttaat tgtccttgtt caaggatgct
     4681 gtctttgaca acagatgttt tcttgccttt gatgttcagc aggaagcttg gcgcaaacgt
     4741 tgattgtttg tctgcgtaga atcctctgtt tgtcatatag cttgtaatca cgacattgtt
     4801 tcctttcgct tgaggtacag cgaagtgtga gtaagtaaag gttacatcgt taggatcaag
     4861 atccattttt aacacaaggc cagttttgtt cagcggcttg tatgggccag ttaaagaatt
     4921 agaaacataa ccaagcatgt aaatatcgtt agacgtaatg ccgtcaatcg tcatttttga
     4981 tccgcgggag tcagtgaaca ggtaccattt gccgttcatt ttaaagacgt tcgcgcgttc
     5041 aatttcatct gttactgtgt tagatgcaat cagcggtttc atcacttttt tcagtgtgta
     5101 atcatcgttt agctcaatca taccgagagc gccgtttgct aactcagccg tgcgtttttt
     5161 atcgctttgc agaagttttt gactttcttg acggaagaat gatgtgcttt tgccatagta
     5221 tgctttgtta aataaagatt cttcgccttg gtagccatct tcagttccag tgtttgcttc
     5281 aaatactaag tatttgtggc ctttatcttc tacgtagtga ggatctctca gcgtatggtt
     5341 gtcgcctgag ctgtagttgc cttcatcgat gaactgctgt acattttgat acgtttttcc
     5401 gtcaccgtca aagattgatt tataatcctc tacaccgttg atgttcaaag agctgtctga
     5461 tgctgatacg ttaacttgtg cagttgtcag tgtttgtttg ccgtaatgtt taccggagaa
     5521 atcagtgtag aataaacgga tttttccgtc agatgtaaat gtggctgaac ctgaccattc
     5581 ttgtgtttgg tcttttagga tagaatcatt tgcatcgaat ttgtcgctgt ctttaaagac
     5641 gcggccagcg tttttccagc tgtcaataga agtttcgccg actttttgat agaacatgta
     5701 aatcgatgtg tcatccgcat ttttaggatc tccggctaat gcaaagacga tgtggtagcc
     5761 gtgatagttt gcgacagtgc cgtcagcgtt ttgtaatggc cagctgtccc aaacgtccag
     5821 gccttttgca gaagagatat ttttaattgt ggacgaatcg aattcaggaa cttgatattt
     5881 ttcatttttt tgctgttcag ggatttgcag catatcatgg cgtgtaatat gggaaatgcc
     5941 gtatgtttcc ttatatggct tttggttcgt ttctttcgca aacgcttgag ttgcgcctcc
     6001 tgccagcagt gcggtagtaa aggttaatac tgttgcttgt tttgcaaact ttttgatgtt
     6061 catcgttcat gtctcctttt ttatgtactg tgttagcggt ctgcttcttc cagccctcct
     6121 gtttgaagat ggcaagttag ttacgcacaa taaaaaaaga cctaaaatat gtaaggggtg
     6181 acgccaaagt atacactttg ccctttacac attttaggtc ttgcctgctt tatcagtaac
     6241 aaacccgcgc gatttacttt tcgacctcat tctattagac tctcgtttgg attgcaactg
     6301 gtctattttc ctcttttgtt tgatagaaaa tcataaaagg atttgcagac tacgggccta
     6361 aagaactaaa aaatctatct gtttcttttc attctctgta ttttttatag tttctgttgc
     6421 atgggcataa agttgccttt ttaatcacaa ttcagaaaat atcataatat ctcatttcac
     6481 taaataatag tgaacggcag gtatatgtga tgggttaaaa aggatcttaa ggcctaggtc
     6541 tagcttggct gcaggggggg gggggggact tccattgttc attccacgga caaaaacaga
     6601 gaaaggaaac gacagaggcc aaaaagcctc gctttcagca cctgtcgttt cctttctttt
     6661 cagagggtat tttaaataaa aacattaagt tatgacgaag aagaacggaa acgccttaaa
     6721 ccggaaaatt ttcataaata gcgaaaaccc gcgaggtcgc cgccccgtaa cctgtcggat
     6781 caccggaaag gacccgtaaa gtgataatga ttatcatcta catatcacaa cgtgcgtgga
     6841 ggccatcaaa ccacgtcaaa taatcaatta tgacgcaggt atcgtattaa ttgatctgca
     6901 tcaacttaac gtaaaaacaa cttcagacaa tacaaatcag cgacactgaa tacggggcaa
     6961 cctcatgtcc cccccccccc ctgcag
//
