LOCUS       Exported                5843 bp ds-DNA     circular SYN 14-AUG-2020
DEFINITION  Pichia pastoris vector for methanol-inducible expression of a 
            secreted protein.
ACCESSION   .
VERSION     .
KEYWORDS    pHpgRNA13
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5843)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 1, 2020 from SnapGene Viewer 5.1.5
            https://www.snapgene.com
COMMENT     Linearize with SacI, PmeI, or BstXI for integration at the AOX1 
            locus.
FEATURES             Location/Qualifiers
     source          1..5843
                     /organism="synthetic DNA construct"
                     /lab_host="Pichia pastoris"
                     /mol_type="other DNA"
     misc_feature    31..135
                     /label=Amp promoter
     CDS             136..996
                     /codon_start=1
                     /label=Amp
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     misc_feature    1154..1765
                     /label=pOpTEF1
     misc_feature    1766..1771
                     /label=N6 spacer
     misc_feature    1772..1814
                     /label=HH
     misc_feature    1815..1834
                     /label=gRNA-ADE12
     misc_feature    1835..1914
                     /label=stgRNA2
     misc_feature    1915..1982
                     /label=HDV
     misc_feature    1983..2323
                     /label=OpAMOtt
     misc_feature    2347..2798
                     /label=panARS
     misc_feature    2805..3452
                     /label=ScLeu2 promoter
     CDS             3453..4547
                     /codon_start=1
                     /label=ScLeu2
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     misc_feature    4548..5024
                     /label=ScLeu2 terminator
     rep_origin      complement(5175..5763)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
       61 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
      121 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
      181 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
      241 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
      301 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
      361 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
      421 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
      481 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
      541 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
      601 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
      661 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
      721 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
      781 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
      841 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
      901 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
      961 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     1021 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     1081 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     1141 cccgtagaaa agatagccct aaagacaccg ttgccataat tagggataaa tctatttagg
     1201 ctccttattt ttcactcggc gcgaaaaaaa atgggcacac ggcgatcgac gagaggtcga
     1261 tcgtctctac gttccagcta catggcagtt ctaagacggg aagtaagatg acactagtag
     1321 atgttttgca aattaggatc acaggcctgc tccagaaaac tttttatcct tcagcggaag
     1381 gtctgtccag gggcacaaat ctcaaacaga gcaaaggcac ccttaccctc ttatctcctt
     1441 gatgaatttt tttttctttt aaattctttt ttagaaggcc gggtaacaac tttaagaacc
     1501 tgtttcagct gtccccagga agctccattg gcagttccag tacgttgtag atgtgtatag
     1561 tgtgctgaaa agcgttgtcc ctggtttttt caagcaaaat cttcgtctcg gagctggata
     1621 gcccaccaag gtattgttcc tgtgcgtaat ttttggcacg cagacgactc gaataagttt
     1681 ggcaataaaa aaattttttt cactatataa agaggagaca ttcccacatg agattttttc
     1741 tgatctttta attagtacat tcgtaatctg actgatgagt ccgtgaggac gaaacgagta
     1801 agctcgtctc agatgcttga aaccccacac gcgtgtttta gagctagaaa tagcaagtta
     1861 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttggccgg
     1921 catggtccca gcctcctcgc tggcgccggc tgggcaacat gcttcggcat ggcgaatggg
     1981 acgtatatag tacacgacaa tctagtaatc tccactattg acgagctcgt cgaactgcga
     2041 aaataggttt tccatctggt ctgtaggcat cagcccggcg tcatcctcct gcgcaggagc
     2101 agcgggctca gggccggcct gggcgggctg atccagaaag tcgaggttca gatctgtgag
     2161 cgagtccatc gcagacggct cgctgttatc tgctgtcttc atctcaaaga tatctttcat
     2221 catatcgtcg ttctgcggcc gctcgctggt ctcgccaaag tccaagaaat ctgtgcggaa
     2281 attgtccaaa tccaggttca tcaggtcgtc tgttgatcca aacgcggcgg ccgccagctt
     2341 tctagatagt gctgattatg atttgacgtt tatatacatg ttatgttaat atggtaaatt
     2401 ttagatattt ggtaggagtg tcggtccttg aaatatatgt aatataccca ttataccaag
     2461 gaaagattga tttaactata ttggattagc agatatctaa tttgtttttt tgttcttaac
     2521 ctctggtgag ttattatgat gtttttatat tttatctttt ttaattaaat gtcattatat
     2581 agaaacaaat attaaactaa ttattaaagt gtattagtat tcttaatgaa cgtcgggaag
     2641 aacaaaagtt taaagatatt gaagttaaac atcaatatta gtcataagtg cataagcaac
     2701 tgatattgat ggaaagaact aaagaagtgc aaagagctca ccaaaaaacg tactgcgtat
     2761 ctgttctttc tcattctgat attatccaaa gatgttgagg atcctcgagg agaacttcta
     2821 gtatatctac atacctaata ttattgcctt attaaaaatg gaatcccaac aattacatca
     2881 aaatccacat tctcttcaaa atcaattgtc ctgtacttcc ttgttcatgt gtgttcaaaa
     2941 acgttatatt tataggataa ttatactcta tttctcaaca agtaattggt tgtttggccg
     3001 agcggtctaa ggcgcctgat tcaagaaata tcttgaccgc agttaactgt gggaatactc
     3061 aggtatcgta agatgcaaga gttcgaatct cttagcaacc attatttttt tcctcaacat
     3121 aacgagaaca cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg aaattttttg
     3181 ttaatttcag aggtcgcctg acgcatatac ctttttcaac tgaaaaattg ggagaaaaag
     3241 gaaaggtgag agcgccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa
     3301 tgcttgcatc acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa
     3361 taggtggtta gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat
     3421 atatatatat atttcaagga tataccattc taatgtctgc ccctaagaag atcgtcgttt
     3481 tgccaggtga ccacgttggt caagaaatca cagccgaagc cattaaggtt cttaaagcta
     3541 tttctgatgt tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg
     3601 ctatcgatgc tacaggtgtt ccacttccag atgaggcgct ggaagcctcc aagaaggctg
     3661 atgccgtttt gttaggtgct gtgggtggtc ctaaatgggg taccggtagt gttagacctg
     3721 aacaaggttt actaaaaatc cgtaaagaac ttcaattgta cgccaactta agaccatgta
     3781 actttgcatc cgactctctt ttagacttat ctccaatcaa gccacaattt gctaaaggta
     3841 ctgacttcgt tgttgtcaga gaattagtgg gaggtattta ctttggtaag agaaaggaag
     3901 acgatggtga tggtgtcgct tgggatagtg aacaatacac cgttccagaa gtgcaaagaa
     3961 tcacaagaat ggccgctttc atggccctac aacatgagcc accattgcct atttggtcct
     4021 tggataaagc taatgttttg gcctcttcaa gattatggag aaaaactgtg gaggaaacca
     4081 tcaagaacga attccctaca ttgaaggttc aacatcaatt gattgattct gccgccatga
     4141 tcctagttaa gaacccaacc cacctaaatg gtattataat caccagcaac atgtttggtg
     4201 atatcatctc cgatgaagcc tccgttatcc caggttcctt gggtttgttg ccatctgcgt
     4261 ccttggcctc tttgccagac aagaacaccg catttggttt gtacgaacca tgccacggtt
     4321 ctgctccaga tttgccaaag aataaggtca accctatcgc cactatcttg tctgctgcaa
     4381 tgatgttgaa attgtcattg aacttgcctg aagaaggtaa ggccattgaa gatgcagtta
     4441 aaaaggtttt ggatgcaggt atcagaactg gtgatttagg tggttccaac agtaccaccg
     4501 aagtcggtga tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt
     4561 ttttatgata tttgtacata aactttataa atgaaattca taatagaaac gacacgaaat
     4621 tacaaaatgg aatatgttca tagggtagac gaaactatat acgcaatcta catacattta
     4681 tcaagaagga gaaaaaggag gatgtaaagg aatacaggta agcaaattga tactaatggc
     4741 tcaacgtgat aaggaaaaag aattgcactt taacattaat attgacaagg aggagggcac
     4801 cacacaaaaa gttaggtgta acagaaaatc atgaaactat gattcctaat ttatatattg
     4861 gaggattttc tctaaaaaaa aaaaaataca acaaataaaa aacactcaat gacctgacca
     4921 tttgatggag tttaagtcaa taccttcttg aaccatttcc cataatggtg aaagttccct
     4981 caagaatttt actctgtcag aaacggcctt aacgacgtag tcgaacgcgt gtacgcatgt
     5041 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
     5101 aagctggaga ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5161 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5221 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5281 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5341 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     5401 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5461 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5521 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5581 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5641 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5701 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5761 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5821 taagggattt tggtcatgag atc
//
