LOCUS       40924_26536        4250 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Intracelluar expression vector pKanB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4250)
  AUTHORS   Lin-Cereghino J, Hashimoto MD, Moy A, Castelo J, Orazem CC, Kuo P, 
            Xiong S, Gandhi V, Hatae CT, Chan A, Lin-Cereghino GP.
  TITLE     Direct selection of Pichia pastoris expression strains using new 
            G418 resistance vectors
  JOURNAL   Yeast 25 (4), 293-299 (2008)
  PUBMED    18327886
REFERENCE   2  (bases 1 to 4250)
  AUTHORS   Lin-Cereghino J, Lin-Cereghino G.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-NOV-2007) Biological Sciences, University of the 
            Pacific, 3601 Pacific Ave, Stockton, CA 95211, USA
REFERENCE   3  (bases 1 to 4250)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4250)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; 
            date: "2008"; volume: "25"; issue: "4"; pages: "293-299"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-NOV-2007) Biological Sciences, University of the Pacific, 3601 
            Pacific Ave, Stockton, CA 95211, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..922
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     misc_feature    921..953
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             971..1000
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             1016..1033
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      1113..1359
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             1918..2730
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2752..2781
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             2797..2814
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2893..3139
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     terminator      3260..3507
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(3582..4170)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa actgcagtct cggatcggta cctcgagccg cggcggccgc
      961 cagctttcta gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca
     1021 tcatcatcat cattgagttt gtagccttag acatgactgt tcctcagttc aagttgggca
     1081 cttacgagaa gaccggtctt gctagattct aatcaagagg atgtcagaat gccatttgcc
     1141 tgagagatgc aggcttcatt tttgatactt ttttatttgt aacctatata gtataggatt
     1201 ttttttgtca ttttgtttct tctcgtacga gcttgctcct gatcagccta tctcgcagct
     1261 gatgaatatc ttgtggtagg ggtttgggaa aatcattcga gtttgatgtt tttcttggta
     1321 tttcccactc ctcttcagag tacagaagat taagtgagac cttcgtttgt gcggatcctt
     1381 gaagctatgg tgtgtgggag ggctaactcg aggtactgat tggacgagga cacaggcctg
     1441 gtgtcctcgt ccaatcaggt agccatctct gaaatatctg gctccgttgc aactccgaac
     1501 gacctgctgg caacgtaaaa ttctccgggg taaaacttaa atgtggagta atggaaccag
     1561 aaacgtctct tcccttctct ctccttccac cgcccgttac cgtccctagg aaattttact
     1621 ctgctggaga gcttcttcta cggccccctt gcagcaatgc tcttcccagc attacgttgc
     1681 gggtaaaacg gaggtcgtgt acccgaccta gcagcccagg gatggaaaag tcccggccgt
     1741 cgctggcaat aatagcgggc ggacgcatgt catgagatta ttggaaacca ccagaatcga
     1801 atataaaagg cgaacacctt tcccaatttt ggtttctcct gacccaaaga ctttaaattt
     1861 aatttatttg tccctatttc aatcaattga acaactattt cgaaacgagg aattcttatg
     1921 agccatattc aacgggaaac gtcttgctcg aggccgcgat taaattccaa catggatgct
     1981 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
     2041 cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
     2101 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
     2161 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
     2221 cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
     2281 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
     2341 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
     2401 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
     2461 atgcataagc ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     2521 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     2581 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     2641 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     2701 cagtttcatt tgatgctcga tgagtttttc taagcggccg ccagctttct agaacaaaaa
     2761 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca tcattgagtt
     2821 ttagccttag acatgactgt tcctcagttc aagttgggca cttacgagaa gaccggtctt
     2881 gctagattct aatcaagagg atgtcagaat gccatttgcc tgagagatgc aggcttcatt
     2941 tttgatactt ttttatttgt aacctatata gtataggatt ttttttgtca ttttgtttct
     3001 tctcgtacga gcttgctcct gatcagccta tctcgcagct gatgaatatc ttgtggtagg
     3061 ggtttgggaa aatcattcga gtttgatgtt tttcttggta tttcccactc ctcttcagag
     3121 tacagaagat taagtgagac cttcgtttgt gcggatcctt gaagctatgg tgtgtgtggg
     3181 agggctaact cgaggtactg attggacgag gacacaggcc tggtcctcgg agatccgtcc
     3241 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctccc
     3301 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     3361 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     3421 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     3481 ttgggacgct cgaaggcttt aatttgcaag ctggagacca acatgtgagc aaaaggccag
     3541 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3601 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3661 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3721 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     3781 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3841 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3901 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3961 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4021 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4081 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4141 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4201 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatc
//
