LOCUS       sequence-43434-e        5550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5550)
  AUTHORS   Sears IB, O'Connor J, Rossanese OW, Glick BS
  TITLE     A versatile set of vectors for constitutive and regulated gene 
            expression in Pichia pastoris.
  JOURNAL   Yeast. 1998 Jun 15;14(8):783-90.
  PUBMED    9675822
REFERENCE   2  (bases 1 to 5550)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Yeast."; 
            date: "1998-06-15"; volume: "14(8)"; pages: "783-90"
COMMENT     Sequence Label: pIB2
FEATURES             Location/Qualifiers
     source          1..5550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        193..669
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     rep_origin      complement(1074..1662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1154..1173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1833..2624,2625..2693))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2456..2475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             2793..5327
                     /codon_start=1
                     /gene="Pichia pastoris HIS4"
                     /product="multifunctional enzyme, required for histidine 
                     biosynthesis"
                     /label=PpHIS4
                     /note="auxotrophic marker for Pichia pastoris"
                     /translation="MTFPLLPAYASVAEFDNSLSLVGKAVFPYAADQLHNLIKFTQSTE
                     LQVNVQVESSVTEDQFEELIDNLLKLYNNGINEVILDLDLAERVVQRMIPGARVIYRTL
                     VDKVASLPANASIAVPFSSPLGDLKSFTNGGSRTVYAFSETAKLVDVTSTVASGIIPII
                     DARQLTTEYELSEDVKKFPVSEILLASLTTDRPDGLFTTLVADSSNYSLGLVYSSKKSI
                     PEAIRTQTGVYQSRRHGLWYKGATSGATQKLLGIELDCDGDCLKFVVEQTGVGFCHLER
                     TSCFGQSKGLRAMEATLWDRKSNAPEGSYTKRLFDDEVLLNAKIREEADELAEAKSKED
                     IAWECADLFYFALVRCAKYGVTLDEVERNLDMKSLKVTRRKGDAKPGYTKEQPKEESKP
                     KEVPSEGRIELCKIDVSKASSQEIEDALRRPIQKTEQIMELVKPIVDNVRQNGDKALLE
                     LTAKFDGVALKTPVLEAPFPEELMQLPDNVKRAIDLSIDNVRKFHEAQLTETLQVETCP
                     GVVCSRFARPIEKVGLYIPGGTAILPSTSLMLGVPAKVAGRKEIVFASPPKKDGTLTPE
                     VIYVAHKVGAKCIVLAGGAQAVAAMAYGTETVPKCDKIFGPGNQFVTAAKMMVQNDTSA
                     LCSIDMPAGPSEVLVIADKYADPDFVASDLLSQAEHGIDSQVILLAVDMTDKELARIED
                     AVHNQAVQLPRVEIVRKCIAHSTTLSVATYEQALEMSNQYAPEHLILQIENASSYVDQV
                     QHAGSVFVGAYSPESCGDYSSGTNHTLPTYGYARQYSGVNTATFQKFITSQDVTPEGLK
                     HIGQAVMDLAAVEGLDAHRNAVKVRMEKLGLI"
     primer_bind     5523..5541
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatagatc cttttttgta gaaatgtctt ggtgtcctcg tccaatcagg tagccatctc
      241 tgaaatatct ggctccgttg caactccgaa cgacctgctg gcaacgtaaa attctccggg
      301 gtaaaactta aatgtggagt aatggaacca gaaacgtctc ttcccttctc tctccttcca
      361 ccgcccgtta ccgtccctag gaaattttac tctgctggag agcttcttct acggccccct
      421 tgcagcaatg ctcttcccag cattacgttg cgggtaaaac ggaagtcgtg tacccgacct
      481 agcagcccag ggatggaaaa gtcccggccg tcgctggcaa taatagcggg cggacgcatg
      541 tcatgagatt attggaaacc accagaatcg aatataaaag gcgaacacct ttcccaattt
      601 tggtttctcc tgacccaaag actttaaatt taatttattt gtccctattt caatcaattg
      661 aacaactatc aagaattcga gctcggtacc cggggatcca ctagtctcga gctgcaggca
      721 tgcaagcttc ttagacatga ctgttcctca gttcaagttg ggcacttacg agaagaccgg
      781 tcttgctaga ttctaatcaa gaggatgtca gaatgccatt tgcctgagag atgcaggctt
      841 catttttgat acttttttat ttgtaaccta tatagtatag gatttttttt gtcattttgt
      901 ttcttctcgt acgagcttgc tcctgatcag cctatctcgc agctgatgaa tatcttgtgg
      961 taggggtttg ggaaaatcat tcgagtttga tgtttttctt ggtatttccc acacatgtga
     1021 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1081 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1141 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1201 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1261 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1321 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1381 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     1441 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     1501 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     1561 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1621 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     1681 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     1741 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     1801 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     1861 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     1921 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     1981 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2041 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2101 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2161 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2221 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2281 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2341 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     2401 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     2461 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     2521 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     2581 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     2641 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     2701 ctttttcaat gatctcctga tgactgactc actgataata aaaatacggc ttcagaattt
     2761 ctcaagacta cactcactgt ccgacttcaa gtatgacatt tcccttgcta cctgcatacg
     2821 caagtgttgc agagtttgat aattccttga gtttggtagg aaaagccgtg tttccctatg
     2881 ctgctgacca gctgcacaac ctgatcaagt tcactcaatc gactgagctt caagttaatg
     2941 tgcaagttga gtcatccgtt acagaggacc aatttgagga gctgatcgac aacttgctca
     3001 agttgtacaa taatggtatc aatgaagtga ttttggacct agatttggca gaaagagttg
     3061 tccaaaggat gatcccaggc gctagggtta tctataggac cctggttgat aaagttgcat
     3121 ccttgcccgc taatgctagt atcgctgtgc ctttttcttc tccactgggc gatttgaaaa
     3181 gtttcactaa tggcggtagt agaactgttt atgctttttc tgagaccgca aagttggtag
     3241 atgtgacttc cactgttgct tctggtataa tccccattat tgatgctcgg caattgacta
     3301 ctgaatacga actttctgaa gatgtcaaaa agttccctgt cagtgaaatt ttgttggcgt
     3361 ctttgactac tgaccgcccc gatggtctat tcactacttt ggtggctgac tcttctaatt
     3421 actcgttggg cctggtgtac tcgtccaaaa agtctattcc ggaggctata aggacacaaa
     3481 ctggagtcta ccaatctcgt cgtcacggtt tgtggtataa aggtgctaca tctggagcaa
     3541 ctcaaaagtt gctgggtatc gaattggatt gtgatggaga ctgcttgaaa tttgtggttg
     3601 aacaaacagg tgttggtttc tgtcacttgg aacgcacttc ctgttttggc caatcaaagg
     3661 gtcttagagc catggaagcc accttgtggg atcgtaagag caatgctcca gaaggttctt
     3721 ataccaaacg gttatttgac gacgaagttt tgttgaacgc taaaattagg gaggaagctg
     3781 atgaacttgc agaagctaaa tccaaggaag atatagcctg ggaatgtgct gacttatttt
     3841 attttgcatt agttagatgt gccaagtacg gtgtgacgtt ggacgaggtg gagagaaacc
     3901 tggatatgaa gtccctaaag gtcactagaa ggaaaggaga tgccaagcca ggatacacca
     3961 aggaacaacc taaagaagaa tccaaaccta aagaagtccc ttctgaaggt cgtattgaat
     4021 tgtgcaaaat tgacgtttct aaggcctcct cacaagaaat tgaagatgcc cttcgtcgtc
     4081 ctatccagaa aacggaacag attatggaat tagtcaaacc aattgtcgac aatgttcgtc
     4141 aaaatggtga caaagccctt ttagaactaa ctgccaagtt tgatggagtc gctttgaaga
     4201 cacctgtgtt agaagctcct ttcccagagg aacttatgca attgccagat aacgttaaga
     4261 gagccattga tctctctata gataacgtca ggaaattcca tgaagctcaa ctaacggaga
     4321 cgttgcaagt tgagacttgc cctggtgtag tctgctctcg ttttgcaaga cctattgaga
     4381 aagttggcct ctatattcct ggtggaaccg caattctgcc ttccacttcc ctgatgctgg
     4441 gtgttcctgc caaagttgct ggtcgcaaag aaattgtttt tgcatctcca cctaagaagg
     4501 atggcaccct taccccagaa gtcatctacg ttgcccacaa ggttggtgct aagtgtatcg
     4561 tgctagcagg aggcgcccag gcagtagctg ctatggctta cggaacagaa actgttccta
     4621 agtgtgacaa aatatttggt ccaggaaacc agttcgttac tgctgccaag atgatggttc
     4681 aaaatgacac atcagccctg tgtagtattg acatgcctgc tgggccttct gaagttctag
     4741 ttattgctga taaatacgct gatccagatt tcgttgcctc agaccttctg tctcaagctg
     4801 aacatggtat tgattcccag gtgattctgt tggctgtcga tatgacagac aaggagcttg
     4861 ccagaattga agatgctgtt cacaaccaag ctgtgcagtt gccaagggtt gaaattgtac
     4921 gcaagtgtat tgcacactct acaaccctat cggttgcaac ctacgagcag gctttggaaa
     4981 tgtccaatca gtacgctcct gaacacttga tcctgcaaat cgagaatgct tcttcttatg
     5041 ttgatcaagt acaacacgct ggatctgtgt ttgttggtgc ctactctcca gagagttgtg
     5101 gagattactc ctccggcacc aaccacactt tgccaacgta cggatatgcc cgtcaataca
     5161 gcggagttaa cactgcaacc ttccagaagt tcatcacttc acaagacgta actcctgagg
     5221 gactgaaaca tattggccaa gcagtgatgg atctggctgc tgttgaaggt ctagatgctc
     5281 accgcaatgc tgttaaggtt cgtatggaga aactgggact tatttaatta tttagagatt
     5341 ttaacttaca tttagattcg atagatcatt attgaagcat ttatcagggt tattgtctca
     5401 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5461 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5521 aaaataggcg tatcacgagg ccctttcgtc
//
