LOCUS       40924_45973        5344 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pVIK165, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5344)
  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  JOURNAL   Unpublished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 5344)
  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 5344)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5344)
  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            "Unpublished"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..5344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    73..78
                     /label=synthetic RBS
                     /note="synthetic RBS"
     CDS             86..799
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1424..2215
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3777..4145)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(4178..4287)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      4931..5287
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     protein_bind    5322..5338
                     /label=lac operator
                     /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)."
ORIGIN
        1 gaattcccgg gagagctcga tatcgcatgc ggtacctcta gaagaagctt gggatccgtc
       61 gacctcgaat tcggaggaaa caaagatgag taaaggagaa gaacttttca ctggagttgt
      121 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
      181 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
      241 actacctgtt ccatggccaa cacttgtcac tactttcact tatggtgttc aatgcttttc
      301 aagataccca gatcatatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta
      361 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
      421 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
      481 agatggaaac attcttggac acaaattgga atacaactat aactcacaca atgtatacat
      541 catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga
      601 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
      661 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
      721 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
      781 catggatgaa ctatacaaat aaatgtccag acttccaatt cccgggccta tcaagcttat
      841 cgataccgtc gacgttactc tttccgttac gggacaccct gtacaccatg aattgagaaa
      901 aaacggcgag atgtactctt ttccgataac gattggcaat aacgtctgga tcggaagtca
      961 tgtggttatt aatccaggcg tcaccatcgg ggataattct gttattggcg cgggtagtat
     1021 cgtcacaaaa gacattccac caaacgtcgt ggcggctggc gttccttgtc gggttattcg
     1081 ccgcaagcac tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc
     1141 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa
     1201 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg
     1261 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt
     1321 tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag
     1381 gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     1441 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     1501 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     1561 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     1621 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     1681 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     1741 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     1801 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     1861 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     1921 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     1981 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     2041 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     2101 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     2161 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     2221 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     2281 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     2341 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc cgcaatggca
     2401 acaacgcgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     2461 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     2521 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     2581 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     2641 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     2701 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2761 tttaattttg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata
     2821 cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc
     2881 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2941 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca
     3001 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     3061 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     3121 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     3181 ggatccagcc gaccaggctt tccacgcccg cgtgccgctc catgtcgttc gcgcggttct
     3241 cggaaacgcg ctgccgcgtt tcgtgattgt cacgctcaag cccgtagtcc cgttcgagcg
     3301 tcgcgcagag gtcagcgagg gcgcggtagg cccgatacgg ctcatggatg gtgtttcggg
     3361 tcgggtgaat cttgttgatg gcgatatgga tgtgcaggtt gtcggtgtcg tgatgcacgg
     3421 cactgacgcg ctgatgctcg gcgaagccaa gcccagcgca gatgcggtcc tcaatcgcgc
     3481 gcaacgtctc cgcgtcgggc ttctctcccg cgcggaagct aaccagcagg tgataggtct
     3541 tgtcggcctc ggaacgggtg ttgccgtgct gggtcgccat cacctcggcc atgacagcgg
     3601 gcagggtgtt tgcctcgcag ttcgtgacgc gcacgtgacc caggcgctcg gtcttgcctt
     3661 gctcgtcggt gatgtacttc accagctccg cgaagtcgct cttcttgatg gagcgcatgg
     3721 ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa aagtcatggc
     3781 tctgccctcg ggcggaccac gcccatcatg accttgccaa gctcgtcctg cttctcttcg
     3841 atcttcgcca gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg
     3901 gtgagccaga gtttcagcag gccgcccagg cggcccaggt cgccattgat gcgggccagc
     3961 tcgcggacgt gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc
     4021 aggtaggccg acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg
     4081 tctggaaggc agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca
     4141 gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga
     4201 ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg
     4261 ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct
     4321 acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa
     4381 atcgctataa tgaccccgaa gcagggttat gcagcggaaa agcgctgctt ccctgctgtt
     4441 ttgtggaata tctaccgact ggaaacaggc aaatgcagga aattactgaa ctgaggggac
     4501 aggcgagaga cgatgccaaa gagctacacc gacgagctgg ccgagtgggt tgaatcccgc
     4561 gcggccaaga agcgccggcg tgatgaggct gcggttgcgt tcctggcggt gagggcggat
     4621 gtcgaggcgg cgttagcgtc cggctatgcg ctcgtcacca tttgggagca catgcgggaa
     4681 acggggaagg tcaagttctc ctacgagacg ttccgctcgc acgccaggcg gcacatcaag
     4741 gccaagcccg ccgatgtgcc cgcaccgcag gccaaggctg cggaacccgc gccggcaccc
     4801 aagacgccgg agccacggcg gccgaagcag gggggcaagg ctgaaaagcc ggcccccgct
     4861 gcggccccga ccggcttcac cttcaaccca acaccggaca aaaaggatcc agcagttcaa
     4921 cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
     4981 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatggc
     5041 taacgtacta agctctcatg tttcacgtac taagctctca tgtttgaaca ataaaattaa
     5101 tataaatcag caacttaaat agcctctaag gttttaagtt ttataagaaa aaaaagaata
     5161 tataaggctt ttaaagcttt taaggtttaa cggttgtgga caacaagcca gggatgtaac
     5221 gcactgagaa gcccttagag cctctcaaag caattttcag tgacacagga acacttaacg
     5281 gctgacatgg gaattccaca tgtggaattc cacatgtgga attgtgagcg gataacaatt
     5341 tgtg
//
