LOCUS       Exported                7732 bp ds-DNA    circular SYN 04-2-2013
DEFINITION  Pichia pastoris integrating vector for low-copy expression in the 
            PichiaPink(TM) system.
ACCESSION   .
VERSION     .
KEYWORDS    pPink-LC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7732)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
COMMENT     Linearize within PpTRP2 for integration.
FEATURES             Location/Qualifiers
     source          1..7732
                     /organism="synthetic DNA construct"
                     /lab_host="Pichia pastoris"
                     /mol_type="other DNA"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /note="AOX1 promoter"
                     /note="inducible promoter, regulated by methanol"
     misc_feature    941..987
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      1013..1262
                     /gene="S. cerevisiae CYC1"
                     /note="CYC1 terminator"
                     /note="transcription terminator for CYC1"
     promoter        1282..1363
                     /gene="Pichia pastoris ADE2"
                     /note="PpADE2 LC promoter"
                     /note="strong 82-bp promoter fragment that requires only 
                     low-copy integration for efficient expression"
     CDS             1364..3055
                     /codon_start=1
                     /gene="Pichia pastoris ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase from 
                     Pichia pastoris"
                     /note="PpADE2"
                     /note="essential for the biosynthesis of purine 
                     nucleotides, including adenine"
                     /translation="MDSQVIGILGGGQLGRMIVEAASRLNIKTVILDDGFSPAKHINAA
                     QDHIDGSFKDEEAIAKLAAKCDVLTVEIEHVNTDALKRVQDRTGIKIYPLPETIELIKD
                     KYLQKEHLIKHNISVTKSQGIESNEKALLLFGEENGFPYLLKSRTMAYDGRGNFVVESK
                     EDISKALEFLKDRPLYAEKFAPFVKELAVMVVRSLEGEVFSYPTVETVHKDNICHIVYA
                     PARVNDTIQKKAQILAENTVKTFPGAGIFGVEMFLLSDGELLVNEIAPRPHNSGHYTID
                     ACVTSQFEAHVRAITGLPMPLDFTKLSTSNTNAIMLNVLGAEKSHGELEFCRRALETPG
                     ASVYLYGKTTRLARKMGHINIIGSSMLEAEQKLEYILEESTHLPSSTVSADTKPLVGVI
                     MGSDSDLPVISKGCDILKQFGVPFEVTIVSAHRTPQRMTRYAFEAASRGIKAIIAGAGG
                     AAHLPGMVAAMTPLPVIGVPVKGSTLDGVDSLHSIVQMPRGVPVATVAINNATNAALLA
                     IRILGTIDHKWQKEMSKYMNAMETEVLGKASNLESEGYESYLKNRL"
     promoter        3282..3431
                     /gene="Pichia pastoris TRP2"
                     /note="PpTRP2 promoter"
                     /note="promoter for Pichia pastoris TRP2"
     CDS             3432..5048
                     /codon_start=1
                     /gene="Pichia pastoris TRP2"
                     /product="anthranilate synthase from Pichia pastoris"
                     /note="PpTRP2"
                     /note="catalyzes the initial step of tryptophan 
                     biosynthesis"
                     /translation="MSVSDSHLATDYFGTQLTKQIHPSAESFLDIVKNDRQSHSPETVP
                     NLYPIYKFIQSDFLTPHLAYIKLTNSSETPSPSFLLESAKNGDTVDRYTFMGHSPRKII
                     KTGPLEGAEVDPLVLLEKELKGTRQAQLPGIPRLSGGAIGYISYDCIKYFEPKTERKLK
                     DVLQLPEAALMLFDTIVAFDNVYQRFQVIGNVSLSVDDSDEAILEKYYKTREEVEKISK
                     VVFDNKTVPYYEQKDIIQGQTFTSNIGQEGYENHVRKLKEHILKGDIFQAVPSQRVARP
                     TSLHPFNIYRHLRTVNPSPYMFYIDYLDFQVVGASPELLVKSDNNNKIITHPIAGTLPR
                     GKTIEEDDNYAKQLKSSLKDRAEHVMLVDLARNDINRVCEPTSTTVDRLLTVERFSHVM
                     HLVSEVSGTLRPNKTRFDAFRSIFPAGTVSGAPKVRAMQLIGELEGEKRGVYAGAVGHW
                     SYDGKSMDTCIALRTMVVKDGVAYLQAGGGIVYDSDPYDEYIETMNKMRSNNNTILEAE
                     KIWTDRLARDENQSESEENDQ"
     rep_origin      complement(5512..6100)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6271..7131)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7132..7236)
                     /gene="bla"
                     /note="AmpR promoter"