LOCUS       40924_26844        9589 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pKLAC1-Gluc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9589)
  AUTHORS   Colussi PA, Ganatra MB, Read JD, Taron CH.
  TITLE     A vector for expression of Gaussia princeps luciferase tagged 
            proteins in the yeast Kluyveromyces lactis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9589)
  AUTHORS   Colussi PA, Ganatra MB, Read JD, Taron CH.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-AUG-2008) Gene Expression Division, New England 
            Biolabs, 240 County Road, Ipswich, MA 01938, USA
REFERENCE   3  (bases 1 to 9589)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9589)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-AUG-2008) Gene Expression Division, New England Biolabs, 240 
            County Road, Ipswich, MA 01938, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    14..331
                     /note="Kluyveromyces lactis alpha mating factor pre-pro 
                     secretion leader"
     misc_feature    290..331
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             332..838
                     /codon_start=1
                     /gene="Gluc"
                     /product="luciferase"
                     /label=Gluc
                     /note="derived from Gaussia princeps"
                     /protein_id="ACI46139.1"
                     /translation="KPTENNEDFNIVAVASNFATTDLDADRGKLPGKKLPLEVLKEMEA
                     NARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESAQGGIGEAIVDIPEIPGF
                     KDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQRCATFASKIQGQVDKIK
                     GAGGD"
     gene            332..838
                     /gene="Gluc"
                     /label=Gluc
     terminator      853..1471
                     /label=LAC4 terminator
                     /note="transcription terminator for Kluyveromyces lactis
                     LAC4"
     promoter        1472..2174
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             2195..3838
                     /label=amdS
                     /note="acetamidase from Aspergillus nidulans"
     promoter        4477..5133
                     /label=LAC4 promoter (5')
                     /note="5' portion of the Kluyveromyces lactis LAC4
                     promoter"
     promoter        complement(5150..5168)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5182..5198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5206..5222)
                     /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(5230..5260)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5275..5296)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5584..6172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6346..7203)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7204..7308)
                     /label=AmpR promoter
     rep_origin      7335..7790
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     7931..7947
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7961..9489
                     /label=LAC4 promoter (3')
                     /note="3' portion of a modified Kluyveromyces lactis LAC4 
                     promoter"
     protein_bind    9495..9511
                     /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 agaagagcta gatctcctag
      301 gggtaccgtc gacggcgcgc ctgcggccgc taagcccacc gagaacaacg aagacttcaa
      361 catcgtggcc gtggccagca acttcgcgac cacggatctc gatgctgatc gcgggaagtt
      421 gcccggcaag aagctgccgc tggaggtgct caaagagatg gaagccaatg cccggaaagc
      481 tggctgcacc aggggctgtc tgatctgcct gtcccacatc aagtgcacgc ccaagatgaa
      541 gaagttcatc ccaggacgct gccacaccta cgaaggcgac aaagagtccg cacagggcgg
      601 cataggcgag gcgatcgtcg acattcctga gattcctggg ttcaaggact tggagcccat
      661 ggagcagttc atcgcacagg tcgatctgtg tgtggactgc acaactggct gcctcaaagg
      721 gcttgccaac gtgcagtgtt ctgacctgct caagaagtgg ctgccgcaac gctgtgcgac
      781 ctttgccagc aagatccagg gccaggtgga caagatcaag ggggccggtg gtgactaacc
      841 ttgaatcgag aatttatact tagataagta tgtacttaca ggtatatttc tatgagatac
      901 tgatgtatac atgcatgata atatttaaac ggttattagt gccgattgtc ttgtgcgata
      961 atgacgttcc tatcaaagca atacacttac cacctattac atgggccaag aaaatatttt
     1021 cgaacttgtt tagaatatta gcacagagta tatgatgata tccgttagat tatgcatgat
     1081 tcattcctac aactttttcg tagcataagg attaattact tggatgccaa taaaaaaaaa
     1141 aaacatcgag aaaatttcag catgctcaga aacaattgca gtgtatcaaa gtaaaaaaaa
     1201 gattttcact acatgttcct tttgaagaaa gaaaatcatg gaacattaga tttacaaaaa
     1261 tttaaccacc gctgattaac gattagaccg ttaagcgcac aacaggttat tagtacagag
     1321 aaagcattct gtggtgttgc cccggacttt cttttgcgac ataggtaaat cgaataccat
     1381 catactatct tttccaatga ctccctaaag aaagactctt cttcgatgtt gtatacgttg
     1441 gagcataggg caagaattgt ggcttgagat catccttttg ttgtttccgg gtgtacaata
     1501 tggacttcct cttttctggc aaccaaaccc atacatcggg attcctataa taccttcgtt
     1561 ggtctcccta acatgtaggt ggcggagggg agatatacaa tagaacagat accagacaag
     1621 acataatggg ctaaacaaga ctacaccaat tacactgcct cattgatggt ggtacataac
     1681 gaactaatac tgtagcccta gacttgatag ccatcatcat atcgaagttt cactaccctt
     1741 tttccatttg ccatctattg aagtaataat aggcgcatgc aacttctttt cttttttttt
     1801 cttttctctc tcccccgttg ttgtctcacc atatccgcaa tgacaaaaaa atgatggaag
     1861 acactaaagg aaaaaattaa cgacaaagac agcaccaaca gatgtcgttg ttccagagct
     1921 gatgaggggt atctcgaagc acacgaaact ttttccttcc ttcattcacg cacactactc
     1981 tctaatgagc aacggtatac ggccttcctt ccagttactt gaatttgaaa taaaaaaaag
     2041 tttgctgtct tgctatcaag tataaataga cctgcaatta ttaatctttt gtttcctcgt
     2101 cattgttctc gttccctttc ttccttgttt ctttttctgc acaatatttc aagctatacc
     2161 aagcatacaa tcaaggaatt ccggatccgc caccatgcct caatcctggg aagaactggc
     2221 cgctgataag cgcgcccgcc tcgcaaaaac catccctgat gaatggaaag tccagacgct
     2281 gcctgcggaa gacagcgtta ttgatttccc aaagaaatcg gggatccttt cagaggccga
     2341 actgaagatc acagaggctt ccgctgcgga tcttgtgtcc aagctggcgg ccggagagtt
     2401 gacctcggtg gaagttacgc tagcattctg taaacgggca gcaatcgccc agcagttaac
     2461 aaactgcgcc cacgagttct tccctgacgc cgctctcgcg caggcaaggg aactcgatga
     2521 atactacgca aagcacaaga gacccgttgg tccactccat ggcctcccca tctctctcaa
     2581 agaccagctt cgagtcaagg gctacgaaac atcaatgggc tacatctcat ggctaaacaa
     2641 gtacgacgaa ggggactcgg ttctgacaac catgctccgc aaagccggtg ccgtcttcta
     2701 cgtcaagacc tctgtcccgc agaccctgat ggtctgcgag acagtcaaca acatcatcgg
     2761 gcgcaccgtc aacccacgca acaagaactg gtcgtgcggc ggcagttctg gtggtgaggg
     2821 tgcgatcgtt gggattcgtg gtggcgtcat cggtgtagga acggatatcg gtggctcgat
     2881 tcgagtgccg gccgcgttca acttcctgta cggtctaagg ccgagtcatg ggcggctgcc
     2941 gtatgcaaag atggcgaaca gcatggaggg tcaggagacg gtgcacagcg ttgtcgggcc
     3001 gattacgcac tctgttgagg acctccgcct cttcaccaaa tccgtcctcg gtcaggagcc
     3061 atggaaatac gactccaagg tcatccccat gccctggcgc cagtccgagt cggacattat
     3121 tgcctccaag atcaagaacg gcgggctcaa tatcggctac tacaacttcg acggcaatgt
     3181 ccttccacac cctcctatcc tgcgcggcgt ggaaactacc gtcgccgcac tcgccaaagc
     3241 cggtcacacc gtgaccccgt ggacgccata caagcacgat ttcggccacg atctcatctc
     3301 ccatatctac gcggctgacg gcagcgccga cgtaatgcgc gatatcagtg catccggcga
     3361 gccggcgatt ccaaatatca aagacctact gaacccgaac atcaaagctg ttaacatgaa
     3421 cgagctctgg gacacgcatc tccagaagtg gaattaccag atggagtacc ttgagaaatg
     3481 gcgggaggct gaagaaaagg ccgggaagga actggacgcc atcatcgcgc cgattacgcc
     3541 taccgctgcg gtacggcatg accagttccg gtactatggg tatgcctctg tgatcaacct
     3601 gctggatttc acgagcgtgg ttgttccggt tacctttgcg gataagaaca tcgataagaa
     3661 gaatgagagt ttcaaggcgg ttagtgagct tgatgccctc gtgcaggaag agtatgatcc
     3721 ggaggcgtac catggggcac cggttgcagt gcaggttatc ggacggagac tcagtgaaga
     3781 gaggacgttg gcgattgcag aggaagtggg gaagttgctg ggaaatgtgg tgactccata
     3841 gcccgggggg ggctcgatcc cctcgcgagt tggttcagct gctgcctgag gctggacgac
     3901 ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc aggaaaccag cagcggctat
     3961 ccgcgcatcc atgcccccga actgcaggag tggggaggca cgatggccgc tttggtcgat
     4021 ctagattacg tggaagaaag gtagtaaaag tagtagtata agtagtaaaa agaggtaaaa
     4081 agagaaaacc ggctacatac tagagaagca cgtacacaaa aactcatagg cacttcatca
     4141 tacgacagtt tcttgatgca ttataatagt gtattagata ttttcagaaa tatgcataga
     4201 acctcctctt gcctttactt tttatacata gaacattggc agatttactt acactacttt
     4261 gtttctacgc catttctttt gttttcaaca cttagacaag ttgttgagaa ccggactact
     4321 aaaaagcaat gttcccactg aaaatcatgt acctgcagga taataacccc ctaattctgc
     4381 atcgatccag tatgtttttt tttctctact catttttacc tgaagataga gcttctaaaa
     4441 caaaaaaaat cagtgattac atgcatattg tgtgttctag taaccaaagg aaaggaacag
     4501 atagataaaa ttccgagact gtcaaattag gtttttttct ttttttttgg cgggagtcag
     4561 tgggccgaaa tatgttcttg gcctagaact taatctggtt tgatcatgcc aatacttgcc
     4621 tgagtgcccg actttttgcc caccctcttg ccttctgtca tccttcaaaa cccacctgtt
     4681 ttccagccgt atcttcgctc gcatctacac atactgtgcc atatcttgtg tgtagccgga
     4741 cgtgactatg accaaaaaca aacaaggaga actgttcgcc gatttgtaac actcctgcat
     4801 ccatccaagt gggtatgcgc tatgcaatgt taagctaggt caggtcagac caggtccaag
     4861 gacagcaact tgactgtatg caacctttac catctttgca cagaacatac ttgtagctag
     4921 ctagttacac ttatggaccg aaaaggcacc ccaccatgtc tgtccggctt tagagtacgg
     4981 ccgcagaccg ctgatttgcc ttgccaagca gtagtcacaa tgcatcgcat gagcacacgg
     5041 gcacgggcac gggcacagga accattggca aaaataccag atacactata ccgacgtata
     5101 tcaagcccaa gtttaaaatt cctaaatttc cgcggctact tttcaattcc ctatagtgag
     5161 tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5221 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5281 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5341 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5401 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5461 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5521 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5581 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5761 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5941 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6001 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6061 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6121 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6181 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6241 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6301 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6361 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6421 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6481 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6541 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6601 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6661 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6721 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6781 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6841 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6901 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6961 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7021 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7081 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7141 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7201 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7261 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7321 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     7381 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     7441 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     7501 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     7561 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     7621 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     7681 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     7741 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca
     7801 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     7861 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     7921 tcacgacgtt gtaaaacgac ggccagtgcc aagctcccgc ggggatcgac tcataaaata
     7981 gtaaccttct aatgcgtatc tattgactac caaccattag tgtggttgca gaaggcggaa
     8041 ttctcccttc ttcgaattca gcttgctttt tcatttttta ttttccattt ttcagttttt
     8101 gtttgtgtcg aatttagcca gttgcttctc caagatgaaa aaaacccctg cgcagtttct
     8161 gtgctgcaag atcctaatcg acttttccac cccccacaaa agtaaatgtt cttttgttac
     8221 attcgcgtgg gtagctagct ccccgaatct tcaaaggact tagggactgc actacatcag
     8281 agtgtgttca cctggtttgc tgcctggttt gaaagaaaag agcagggaac tcgcgggttc
     8341 ccggcgaata atcatgcgat agtcctttgg ccttccaagt cgcatgtaga gtagacaaca
     8401 gacagggagg gcaggaagga tctttcactg agatcctgta tcttgttggg taagtcggat
     8461 gaaaggggaa tcgtatgaga ttggagagga tgcggaagag gtaacgcctt ttgttaactt
     8521 gtttaattat tatggggcag gcgagagggg gaggaatgta tgtgtgtgag gcgggcgaga
     8581 cggagccatc caggccaggt agaaatagag aaagccgaat gttagacaat atggcagcgt
     8641 agtagagtag gtaggtaggc aagtactgct agcaaagagg agaagggtaa gctcactctt
     8701 cgcattccac accgttagtg tgtcagtttg aacaaaaaaa caatcatcat accaattgat
     8761 ggactgtgga ctggcttttg gaacggcttt tcggactgcg attattcgtg aggaatcaag
     8821 gtaggaattt ggtcatattt acggacaaca gtgggtgatt cccatatgga gtaggaaaac
     8881 gagatcatgg tatcctcaga tatgttgcgg aattctgttc accgcaaagt tcagggtgct
     8941 ctggtgggtt tcggttggtc tttgctttgc ttctcccttg tcttgcatgt taataatagc
     9001 ctagcctgtg agccgaaact tagggtaggc ttagtgttgg aacgtacata tgtatcacgt
     9061 tgacttggtt taaccaggcg acctggtagc cagccatacc cacacacgtt ttttgtatct
     9121 tcagtatagt tgtgaaaagt gtagcggaaa tttgtggtcc gagcaacagc gtctttttct
     9181 agtagtgcgg tcggttactt ggttgacatt ggtatttgga ctttgttgct acaccattca
     9241 ctacttgaag tcgagtgtga agggtatgat ttctagtggt gaacaccttt agttacgtaa
     9301 tgttttcatt gctgttttac ttgagatttc gattgagaaa aaggtattta atagctcgaa
     9361 tcaatgtgag aacagagaga agatgttctt ccctaactcg aaaggtatat gaggcttgtg
     9421 tttcttagga gaattattat tcttttgtta tgttgcgctt gtagttggaa aaggtgaaga
     9481 gacaaaagct ggaattgtga gcggataaca agctcaacac ttgaaattta ggaaagagca
     9541 gaatttggca aaaaaaataa aaaaaaaata aacacacata ctcatcgag
//
