LOCUS       pPink-LC.        7732 bp DNA     circular SYN 01-JAN-1980
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
REFERENCE   2  (bases 1 to 7732)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Linearize within PpTRP2 for integration.
FEATURES             Location/Qualifiers
     source          1..7732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     misc_feature    941..987
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      1013..1262
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     promoter        1282..1363
                     /label=PpADE2 LC promoter
                     /note="strong 82-bp promoter fragment that requires only 
                     low-copy integration for efficient expression"
     CDS             1364..3052
                     /label=PpADE2
                     /note="phosphoribosylaminoimidazole carboxylase from Pichia
                     pastoris"
     promoter        3282..3431
                     /label=PpTRP2 promoter
                     /note="promoter for Pichia pastoris TRP2"
     CDS             3432..5045
                     /label=PpTRP2
                     /note="anthranilate synthase from Pichia pastoris"
     primer_bind     complement(5110..5126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5134..5150)
                     /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(5158..5188)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5203..5224)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5512..6100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6274..7131)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7132..7236)
                     /label=AmpR promoter
     primer_bind     7710..7726
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 acaactaatt attcgaaacg gaattccgga ccggcatgca
      961 ggcctggtac cggccggcca tttaaataca ggcccctttt cctttgtcga tatcatgtaa
     1021 ttagttatgt cacgcttaca ttcacgccct cctcccacat ccgctctaac cgaaaaggaa
     1081 ggagttagac aacctgaagt ctaggtccct atttattttt tttaatagtt atgttagtat
     1141 taagaacgtt atttatattt caaatttttc ttttttttct gtacaaacgc gtgtacgcat
     1201 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     1261 gcaagctgga tccgcggccg cgacttgact cttcactagc ctatgcaaat aaggttacct
     1321 tttccaagaa tcgtagaaac gattaaaaaa cttccaaact ctcatggatt ctcaggtaat
     1381 aggtattcta ggaggaggcc agctaggccg aatgattgtt gaggccgcta gcaggctcaa
     1441 tatcaagacc gtgattcttg atgatggttt ttcacctgct aagcacatta atgctgcgca
     1501 agaccacatc gacggatcat tcaaagatga ggaggctatc gccaagttag ctgccaaatg
     1561 tgatgttctc actgtagaga ttgagcatgt caacacagat gctctaaaga gagttcaaga
     1621 cagaactgga atcaagatat atcctttacc agagacaatc gaactaatca aggataagta
     1681 cttgcaaaag gaacatttga tcaagcacaa catttcggtg acaaagtctc agggtataga
     1741 atctaatgaa aaggcgctgc ttttgtttgg agaagagaat ggatttccat atctgttgaa
     1801 gtcccggact atggcttatg atggaagagg caattttgta gtggagtcta aagaggacat
     1861 cagtaaggca ttagagttct tgaaagatcg tccattgtat gccgagaagt ttgctccttt
     1921 tgttaaagaa ttagcggtaa tggttgtgag atcactggaa ggcgaagtat tctcctaccc
     1981 aaccgtagaa actgtgcaca aggacaatat ctgtcatatt gtgtatgctc cggccagagt
     2041 taatgacacc atccaaaaga aagctcaaat attagctgaa aacactgtga agactttccc
     2101 aggcgctgga atcttcggag ttgagatgtt cctattgtct gatggagaac ttcttgtaaa
     2161 tgagattgct ccaaggcccc acaattctgg tcactataca atcgatgcat gtgtaacatc
     2221 tcagttcgaa gcacatgtaa gagccataac tggtctgcca atgccactag atttcaccaa
     2281 actatctact tccaacacca acgctattat gctcaatgtt ttgggtgctg aaaaatctca
     2341 cggggaatta gagttttgta gaagagcctt agaaacaccc ggtgcttctg tatatctgta
     2401 cggaaagacc acccgattgg ctcgtaagat gggtcatatc aacataatag gatcttccat
     2461 gttggaagca gaacaaaagt tagagtacat tctagaagaa tcaacccact taccatccag
     2521 tactgtatca gctgacacta aaccgttggt tggagttatc atgggttcag actctgatct
     2581 acctgtgatt tcgaaaggtt gcgatatttt aaaacagttt ggtgttccat tcgaagttac
     2641 tattgtctct gctcatagaa caccacagag aatgaccaga tatgcctttg aagccgctag
     2701 tagaggtatc aaggctatca ttgcaggtgc tggtggtgct gctcatcttc caggaatggt
     2761 tgctgccatg actccgttgc cagtcattgg tgttcctgtc aagggctcta cgttggatgg
     2821 tgtagactcg ctacactcga ttgtccaaat gcctagaggt gttcctgtgg ctacggttgc
     2881 tatcaacaac gccaccaatg ccgctctgtt ggccatcagg attttaggta caattgacca
     2941 caaatggcaa aaggaaatgt ccaagtatat gaatgcaatg gagaccgaag tgttggggaa
     3001 ggcatccaac ttggaatctg aagggtatga atcctatttg aagaatcgtc tttgaattta
     3061 gtattgtttt ttaatagatg tatatataat agtacacgta acttatctat tccattcata
     3121 attttatttt aaaggttcgg tagaaatttg tcctccaaaa agttggttag agcctggcag
     3181 ttttgatagg cattattata gattgggtaa tatttaccct gcacctggag gaactttgca
     3241 aagagcctca tgtgcggcgc gccaggccat aatggccaaa cggtttctca attactatat
     3301 actactaacc atttacctgt agcgtatttc ttttccctct tcgcgaaagc tcaagggcat
     3361 cttcttgact catgaaaaat atctggattt cttctgacag atcatcaccc ttgagcccaa
     3421 ctctctagcc tatgagtgta agtgatagtc atcttgcaac agattatttt ggaacgcaac
     3481 taacaaagca gatacaccct tcagcagaat cctttctgga tattgtgaag aatgatcgcc
     3541 aaagtcacag tcctgagaca gttcctaatc tttaccccat ttacaagttc atccaatcag
     3601 acttcttaac gcctcatctg gcttatatca agcttaccaa cagttcagaa actcccagtc
     3661 caagtttctt gcttgaaagt gcgaagaatg gtgacaccgt tgacaggtac acctttatgg
     3721 gacattcccc cagaaaaata atcaagactg ggcctttaga gggtgctgaa gttgacccct
     3781 tggtgcttct ggaaaaagaa ctgaagggca ccagacaagc gcaacttcct ggtattcctc
     3841 gtctaagtgg tggtgccata ggatacatct cgtacgattg tattaagtac tttgaaccaa
     3901 aaactgaaag aaaactgaaa gatgttttgc aacttccgga agcagctttg atgttgttcg
     3961 acacgatcgt ggcttttgac aatgtttatc aaagattcca ggtaattgga aacgtttctc
     4021 tatccgttga tgactcggac gaagctattc ttgagaaata ttataagaca agagaagaag
     4081 tggaaaagat cagtaaagtg gtatttgaca ataaaactgt tccctactat gaacagaaag
     4141 atattattca aggccaaacg ttcacctcta atattggtca ggaagggtat gaaaaccatg
     4201 ttcgcaagct gaaagaacat attctgaaag gagacatctt ccaagctgtt ccctctcaaa
     4261 gggtagccag gccgacctca ttgcaccctt tcaacatcta tcgtcatttg agaactgtca
     4321 atccttctcc atacatgttc tatattgact atctagactt ccaagttgtt ggtgcttcac
     4381 ctgaattact agttaaatcc gacaacaaca acaaaatcat cacacatcct attgctggaa
     4441 ctcttcccag aggtaaaact atcgaagagg acgacaatta tgctaagcaa ttgaagtcgt
     4501 ctttgaaaga cagggccgag cacgtcatgc tggtagattt ggccagaaat gatattaacc
     4561 gtgtgtgtga gcccaccagt accacggttg atcgtttatt gactgtggag agattttctc
     4621 atgtgatgca tcttgtgtca gaagtcagtg gaacattgag accaaacaag actcgcttcg
     4681 atgctttcag atccattttc ccagcaggaa ccgtctccgg tgctccgaag gtaagagcaa
     4741 tgcaactcat aggagaattg gaaggagaaa agagaggtgt ttatgcgggg gccgtaggac
     4801 actggtcgta cgatggaaaa tcgatggaca catgtattgc cttaagaaca atggtcgtca
     4861 aggacggtgt cgcttacctt caagccggag gtggaattgt ctacgattct gacccctatg
     4921 acgagtacat cgaaaccatg aacaaaatga gatccaacaa taacaccatc ttggaggctg
     4981 agaaaatctg gaccgatagg ttggccagag acgagaatca aagtgaatcc gaagaaaacg
     5041 atcaatgaac ggaggacgta agtaggaatt tatggtttgg ccataatggc ctagcttggc
     5101 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5161 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5221 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5941 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6061 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6181 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6241 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6301 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6361 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6421 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6481 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6541 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6601 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6661 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6721 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6781 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6841 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6901 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6961 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7021 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7081 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7141 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7201 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7261 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7321 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     7381 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     7441 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     7501 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     7561 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     7621 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7681 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tg
//
