LOCUS       40924_26854        9107 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pKLAC2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9107)
  AUTHORS   Colussi PA, Read JD, Taron CH.
  TITLE     An integrative expression vector for the yeast Kluyveromyces lactis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9107)
  AUTHORS   Colussi PA, Read JD, Taron CH.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-OCT-2007) Gene Expression Division, New England 
            Biolabs, 240 County Road, Ipswich, MA 01938, USA
REFERENCE   3  (bases 1 to 9107)
  AUTHORS   Colussi PA, Read JD, Taron CH.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-AUG-2008) Gene Expression Division, New England 
            Biolabs, 240 County Road, Ipswich, MA 01938, USA
REFERENCE   4  (bases 1 to 9107)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 9107)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-OCT-2007) Gene Expression Division, New England Biolabs, 240 
            County Road, Ipswich, MA 01938, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (14-AUG-2008) Gene Expression Division, New England Biolabs, 240 
            County Road, Ipswich, MA 01938, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     On Aug 14, 2008 this sequence version replaced EU196354.1.
FEATURES             Location/Qualifiers
     source          1..9107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..349
                     /note="Kluyveromyces lactis alpha mating factor pre-pro 
                     secretion leader"
     misc_feature    257..354
                     /label=multiple cloning site
                     /note="multiple cloning site"
     terminator      371..989
                     /label=LAC4 terminator
                     /note="transcription terminator for Kluyveromyces lactis
                     LAC4"
     promoter        990..1692
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1713..3356
                     /label=amdS
                     /note="acetamidase from Aspergillus nidulans"
     promoter        3995..4651
                     /label=LAC4 promoter (5')
                     /note="5' portion of the Kluyveromyces lactis LAC4
                     promoter"
     promoter        complement(4668..4686)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4700..4716)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4724..4740)
                     /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)."
     promoter        complement(4748..4778)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4793..4814)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5102..5690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5864..6721)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6722..6826)
                     /label=AmpR promoter
     rep_origin      6853..7308
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     7449..7465
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7479..9007
                     /label=LAC4 promoter (3')
                     /note="3' portion of a modified Kluyveromyces lactis LAC4 
                     promoter"
     protein_bind    9013..9029
                     /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 aagcttgaaa aaaatgaaat tctctactat attagccgca tctactgctt taatttccgt
       61 tgttatggct gctccagttt ctaccgaaac tgacatcgac gatcttccaa tatcggttcc
      121 agaagaagcc ttgattggat tcattgactt aaccggggat gaagtttcct tgttgcctgt
      181 taataacgga acccacactg gtattctatt cttaaacacc accatcgctg aagctgcttt
      241 cgctgacaag gatgatctcg agaaaagaga ggctgaagct agaagagctc atatgtccat
      301 gggcggccgc gatatcgtcg acggatccga attccctgca ggtaattaaa taaaggcctt
      361 gaatcgagaa tttatactta gataagtatg tacttacagg tatatttcta tgagatactg
      421 atgtatacat gcatgataat atttaaacgg ttattagtgc cgattgtctt gtgcgataat
      481 gacgttccta tcaaagcaat acacttacca cctattacat gggccaagaa aatattttcg
      541 aacttgttta gaatattagc acagagtata tgatgttatc cgttagatta tgcatgattc
      601 attcctacaa ctttttcgta gcataaggat taattacttg gatgccaata aaaaaaaaaa
      661 acatcgagaa aatttcagca tgctcagaaa caattgcagt gtatcaaagt aaaaaaaaga
      721 ttttcactac atgttccttt tgaagaaaga aaatcatgga acattagatt tacaaaaatt
      781 taaccaccgc tgattaacga ttagaccgtt aagcgcacaa caggttatta gtacagagaa
      841 agcattctgt ggtgttgccc cggactttct tttgcgacat aggtaaatcg aataccatca
      901 tactatcttt tccaatgact ccctaaagaa agactcttct tcgatgttgt atacgttgga
      961 gcatagggca agaattgtgg cttgagatca tccttttgtt gtttccgggt gtacaatatg
     1021 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg
     1081 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac
     1141 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga
     1201 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt
     1261 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct
     1321 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac
     1381 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga
     1441 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc
     1501 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt
     1561 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca
     1621 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa
     1681 gcatacaatc aagcaattcc agatctgcca ccatgcctca atcctgggaa gaactggccg
     1741 ctgataagcg cgcccgcctc gcaaaaacca tccctgatga atggaaagtc cagacgctgc
     1801 ctgcggaaga cagcgttatt gatttcccaa agaaatcggg catcctttca gaggccgaac
     1861 tgaagatcac agaggcttcc gctgcggatc ttgtgtccaa gctggcggcc ggagagttga
     1921 cctcggtgga agttacgcta gcattctgta aacgggcagc aatcgcccag cagttaacaa
     1981 actgcgccca cgagttcttc cctgacgccg ctctcgcgca ggcaagggaa ctcgatgaat
     2041 actacgcaaa gcacaagaga cccgttggtc cacttcatgg cctccccatc tctctcaaag
     2101 accagcttcg agtcaagggc tacgaaacat caatgggcta catctcatgg ctaaacaagt
     2161 acgacgaagg ggactcggtt ctgacaacca tgctccgcaa agccggtgcc gtcttctacg
     2221 tcaagacctc tgtcccgcag accctgatgg tctgcgagac agtcaacaac atcatcgggc
     2281 gcaccgtcaa cccacgcaac aagaactggt cgtgcggcgg cagttctggt ggtgagggtg
     2341 cgatcgttgg gattcgtggt ggcgtcatcg gtgtaggaac ggacatcggt ggctcgattc
     2401 gagtgccggc cgcgttcaac ttcctgtacg gtctaaggcc gagtcatggg cggctgccgt
     2461 atgcaaagat ggcgaacagc atggagggtc aggagacggt gcacagcgtt gtcgggccga
     2521 ttacgcactc tgttgaggac ctccgcctct tcaccaaatc cgtcctcggt caggagcctt
     2581 ggaaatacga ctccaaggtc atccccatgc cctggcgcca gtccgagtcg gacattattg
     2641 cctccaagat caagaacggc gggctcaata tcggctacta caacttcgac ggcaatgtcc
     2701 ttccacaccc tcctatcctg cgcggcgtgg aaactaccgt cgccgcactc gccaaagccg
     2761 gtcacaccgt gaccccgtgg acgccataca agcacgattt cggccacgat ctcatctccc
     2821 atatctacgc ggctgacggc agcgccgacg taatgcgcga catcagtgca tccggcgagc
     2881 cggcgattcc aaatatcaaa gacctactga acccgaacat caaagctgtt aacatgaacg
     2941 agctctggga cacgcatctc cagaagtgga attaccagat ggagtacctt gagaaatggc
     3001 gggaggctga agaaaaggcc gggaaggaac tggacgccat catcgcgccg attacgccta
     3061 ccgctgcggt acggcatgac cagttccggt actatgggta tgcctctgtg atcaacctgc
     3121 tggatttcac gagcgtggtt gttccggtta cctttgcgga taagaacatc gataagaaga
     3181 atgagagttt caaggcggtt agtgagcttg atgccctcgt gcaggaagag tatgatccgg
     3241 aggcgtatca tggggcaccg gttgcagtgc aggttatcgg acggagactc agtgaagaga
     3301 ggacgttggc gattgcagag gaagtgggga agttgctggg aaatgtggtg actccatagc
     3361 ccgggggggg ctcgatcccc tcgcgagttg gttcagctgc tgcctgaggc tggacgacct
     3421 cgcggagttc taccggcagt gcaaatccgt cggcatccag gaaaccagca gcggctatcc
     3481 gcgcatccat gcccccgaac tgcaggagtg gggaggcacg atggccgctt tggtcgatct
     3541 agattacgtg gaagaaaggt agtaaaagta gtagtataag tagtaaaaag aggtaaaaag
     3601 agaaaaccgg ctacatacta gagaagcacg tacacaaaaa ctcataggca cttcatcata
     3661 cgacagtttc ttgatgcatt ataatagtgt attagatatt ttcagaaata tgcatagaac
     3721 ctcctcttgc ctttactttt tatacataga acattggcag atttacttac actactttgt
     3781 ttctacgcca tttcttttgt tttcaacact tagacaagtt gttgagaacc ggactactaa
     3841 aaagcaatgt tcccactgaa aatcatgtac ctgcagcata ataaccccct aattctgcat
     3901 cgatccagta tgtttttttt tctctactca tttttacctg aagatagagc ttctaaaaca
     3961 aaaaaaatca gtgattacat gcatattgtg tgttctagta accaaaggaa aggaacagat
     4021 agataaaatt ccgagactgt caaattaggt ttttttcttt ttttttggcg ggagtcagtg
     4081 ggccgaaata tgttcttggc ctagaactta atctggtttg atcatgccaa tacttgcctg
     4141 agtgcccgac tttttgccca ccctcttgcc ttctgtcatc cttcaaaacc cacctgtttt
     4201 ccagccgtat cttcgctcgc atctacacat actgtgccat atcttgtgtg tagccggacg
     4261 tgactatgac caaaaacaaa caaggagaac tgttcgccga tttgtaacac tcctgcatcc
     4321 atccaagtgg gtatgcgcta tgcaatgtta agctaggtca ggtcagacca ggtccaagga
     4381 cagcaacttg actgtatgca acctttacca tctttgcaca gaacatactt gtagctagct
     4441 agttacactt atggaccgaa aaggcacccc accatgtctg tccggcttta gagtacggcc
     4501 gcagaccgct gatttgcctt gccaagcagt agtcacaatg catcgcatga gcacacgggc
     4561 acgggcacgg gcacaggaac cattggcaaa aataccagat acactatacc gacgtatatc
     4621 aagcccaagt ttaaaattcc taaatttccg cggctacttt tcaattccct atagtgagtc
     4681 gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4741 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4801 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4861 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4921 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4981 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5041 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5101 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5161 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5221 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5281 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5341 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5401 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5461 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5521 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5581 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5641 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5701 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5761 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5821 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5881 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5941 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6001 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6061 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6121 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6181 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6241 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6301 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6361 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6421 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6481 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6541 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6601 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6661 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6721 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6781 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6841 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     6901 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6961 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     7021 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     7081 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     7141 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     7201 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     7261 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt
     7321 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     7381 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7441 acgacgttgt aaaacgacgg ccagtgccaa gctcccgcgg ggatcgactc ataaaatagt
     7501 aaccttctaa tgcgtatcta ttgactacca accattagtg tggttgcaga aggcggaatt
     7561 ttcccttctt cgaatttagc ttgctttttc attttttatt ttccattttt cagtttttgt
     7621 ttgtgtcgaa tttagccagt tgcttctcca agatgaaaaa aacccctgcg cagtttctgt
     7681 gctgcaagat cctaatcgac ttttccaccc cccacaaaag taaatgttct tttgttacat
     7741 tcgcgtgggt agctagctcc ccgaatcttc aaaggactta gggactgcac tacatcagag
     7801 tgtgttcacc tggtttgctg cctggtttga aagaaaagag cagggaactc gcgggttccc
     7861 ggcgaataat catgcgatag tcctttggcc ttccaagtcg catgtagagt agacaacaga
     7921 cagggagggc aggaaggatc tttcactgag atcctgtatc ttgttgggta agtcggatga
     7981 aaggggaatc gtatgagatt ggagaggatg cggaagaggt aacgcctttt gttaacttgt
     8041 ttaattatta tggggcaggc gagaggggga ggaatgtatg tgtgtgaggc gggcgagacg
     8101 gagccatcca ggccaggtag aaatagagaa agccgaatgt tagacaatat ggcagcgtag
     8161 tagagtaggt aggtaggcaa gtactgctag caaagaggag aagggtaagc tcactcttcg
     8221 cattccacac cgttagtgtg tcagtttgaa caaaaaaaca atcatcatac caattgatgg
     8281 actgtggact ggcttttgga acggcttttc ggactgcgat tattcgtgag gaatcaaggt
     8341 aggaatttgg tcatatttac ggacaacagt gggtgattcc catatcgagt aggaaaacga
     8401 gatcatggta tcctcagata tgttgcggaa atctgttcac cgcaaagttc agggtgctct
     8461 ggtgggtttc ggttggtctt tgctttgctt ctcccttgtc ttgcatgtta ataatagcct
     8521 agcctgtgag ccgaaactta gggtaggctt agtgttggaa cgtacatatc tatcacgttg
     8581 acttggttta accaggcgac ctggtagcca gccataccca cacacgtttt ttgtatcttc
     8641 agtatagttg tgaaaagtgt agcggaaatt tgtggtccga gcaacagcgt ctttttctag
     8701 tagtgcggtc ggttacttgg ttgacattgg tatttggact ttgttgctac accattcact
     8761 acttgaagtc gagtgtgaag ggtatgattt ctagtggtga acacctttag ttacgtaatg
     8821 ttttcattgc tgttttactt gagatttcga ttgagaaaaa ggtatttaat agctcgaatc
     8881 aatgtgagaa cagagagaag atgttcttcc ctaactcgaa aggtatatga ggcttgtgtt
     8941 tcttaggaga attattattc ttttgttatg ttgcgcttgt agttggaaaa ggtgaagaga
     9001 caaaagctgg aattgtgagc ggataacaag ctcaacactt gaaatttagg aaagagcaga
     9061 atttggcaaa aaaaataaaa aaaaaataaa cacacatact catcgag
//
