LOCUS       40924_26724        3555 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKILHIS-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3555)
  AUTHORS   Van Reeth T, Dreze PL, Szpirer J, Szpirer C, Gabant P.
  TITLE     Positive selection vectors to generate fused genes for the 
            expression of his-tagged proteins
  JOURNAL   BioTechniques 25 (5), 898-904 (1998)
  PUBMED    9821593
REFERENCE   2  (bases 1 to 3555)
  AUTHORS   Van Reeth T, Gabant P, Dreze P, Szpirer J, Szpirer C.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-OCT-1997) Departement de Biologie Moleculaire 
            Laboratoire de Biologie du Developpement, Universite Libre de 
            Bruxelles, 67, Rue des Chevaux, 1640 Rhode Saint-Genese, Belgium
REFERENCE   3  (bases 1 to 3555)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3555)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1998"; volume: "25"; issue: "5"; pages: 
            "898-904"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-OCT-1997) Departement de Biologie Moleculaire Laboratoire de 
            Biologie du Developpement, Universite Libre de Bruxelles, 67, Rue 
            des Chevaux, 1640 Rhode Saint-Genese, Belgium"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        290..308
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             338..360
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             380..397
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             401..433
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     misc_feature    458..470
                     /gene="ccdB"
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             626..925
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     terminator      1010..1057
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1153..1532
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1583..2374)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      2786..3374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcga ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat
      241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tcgcgaaatt aatacgactc
      301 actataggga gaccacaacg gtttccctct agataatttt gtttaacttt aagaaggaga
      361 tatacatatg cggggttctc atcatcatca tcatcatggt atggctagca tgactggtgg
      421 acagcaaatg ggtcgggatc tgtacgacga tgacgatatc ggatccgtcg acctggccgt
      481 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      541 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      601 acagttgcgc agcctatacg tacggcagtt taaggtttac acctataaaa gagagagccg
      661 ttatcgtctg tttgtggatg tacagagtga tattattgac acgccggggc gacggatggt
      721 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
      781 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
      841 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
      901 cattaacctg atgttctggg gaatataaat gtcagaattc gaagcttgat ccggctgcta
      961 acaaagcccg aaaggaagct gagttggctg ctgcaccgct gagcaataac tagcataacc
     1021 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1081 atctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     1141 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc gcgggtgtgg tggttacgcg
     1201 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1261 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1321 gttccgattt agagctttac ggcacctcga ccgcaaaaaa cttgatttgg gtgatggttc
     1381 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1441 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatcg cggtctattc
     1501 ttttgattta taagggatgt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1561 acaaaaattt taacaaaatt cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     1621 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     1681 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     1741 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     1801 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg
     1861 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     1921 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     1981 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     2041 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     2101 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     2161 tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt
     2221 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     2281 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     2341 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     2401 gatcagagct tgatcccctg cgccatcaga tccttggcgg cgagaaagcc atccagttta
     2461 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc ggttcgcttg
     2521 ctgtccataa aaccgcccag tctagctatc gccatgtaag cccactgcaa gctacctgct
     2581 ttctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     2641 gtcagcaccg tttctgcgga ctggctttct acgtgaaaag gatctaggtg aagatccttt
     2701 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2761 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2821 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2881 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2941 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3001 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3061 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3121 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3181 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3241 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3301 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3361 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctgggc ttttgctggc
     3421 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3481 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3541 gcgaggaagc ggaag
//
