LOCUS       Exported                9589 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pKLAC1-Gluc, complete sequence.
ACCESSION   FJ010198
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9589
                     /organism="Expression vector pKLAC1-Gluc"
                     /mol_type="other DNA"
                     /db_xref="taxon:568092"
     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"
     regulatory      853..1435
                     /regulatory_class="terminator"
                     /note="Kluyveromyces lactis LAC4 transcription terminator"
     regulatory      1492..2194
                     /regulatory_class="promoter"
                     /note="Saccharomyces cerevisiae ADH1 promoter"
     CDS             2195..3841
                     /codon_start=1
                     /gene="amdS"
                     /product="acetamidase"
                     /label=amdS
                     /note="derived from Aspergillus nidulans; confers ability 
                     to process acetamide for selection in Kluyveromyces lactis"
                     /protein_id="ACI46138.1"
                     /translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGIL
                     SEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALAQA
                     RELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTMLRK
                     AGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGVIGVG
                     TDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVEDLRLFT
                     KSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPPILRGVE
                     TTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAIPNIKDLL
                     NPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPTAAVRHDQF
                     RYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYDPEAYHGAPV
                     AVQVIGRRLSEERTLAIAEEVGKLLGNVVTP"
     gene            2195..3841
                     /gene="amdS"
                     /label=amdS
     promoter        4477..5133
                     /label=LAC4 promoter (5)
                     /note="5 portion of the Kluyveromyces lactis LAC4 
                     promoter"
     regulatory      4550..5130
                     /regulatory_class="promoter"
                     /note="modified Kluyveromyces lactis LAC4 promoter 5 end"
     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    5206..5222
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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    5275..5296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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(6343..7203)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="confers ampicillin resistance in Escherichia coli"
                     /protein_id="ACI46140.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6343..7203)
                     /gene="bla"
                     /label=bla
     promoter        complement(7204..7308)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7335..7790
                     /direction=RIGHT
                     /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"
     regulatory      7957..9589
                     /regulatory_class="promoter"
                     /note="modified Kluyveromyces lactis LAC4 promoter 3 end"
     promoter        7961..9489
                     /label=LAC4 promoter (3)
                     /note="3 portion of a modified Kluyveromyces lactis LAC4 
                     promoter"
     protein_bind    9495..9511
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."