LOCUS       sequence-43430-e        5798 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 5798)
  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 5798)
  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: pIB4
FEATURES             Location/Qualifiers
     source          1..5798
                     /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"
     rep_origin      complement(1322..1910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1402..1421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2081..2872,2873..2941))
                     /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     2704..2723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3041..5575
                     /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     5771..5789
                     /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 accatacgct cattccaatt ccttctatta ggctactaac accatgactt tattagcctg
      241 tctatcctgg cccccctggc gaggttcatg tttgtttatt tccgaatgca acaagctccg
      301 cattacaccc gaacatcact ccagatgagg gctttctgag tgtggggtca aatagtttca
      361 tgttccccaa atggcccaaa actgacagtt taaacgctgt cttggaacct aatatgacaa
      421 aagcgtgatc tcatccaaga tgaactaagt ttggttcgtt gaaatgctaa cggccagttg
      481 gtcaaaaaga aacttccaaa agtcggcata ccgtttgtct tgtttggtat tgattgacga
      541 atgctcaaaa ataatctcat taatgcttag cgcagtctct ctatcgcttc tgaaccccgg
      601 tgcacctgtg ccgaaacgca aatggggaaa cacccgcttt ttggatgatt atgcattgtc
      661 tccacattgt atgcttccaa gattctggtg ggaatactgc tgatagccta acgttcatga
      721 tcaaaattta actgttctaa cccctacttg acagcaatat ataaacagaa ggaagctgcc
      781 ctgtcttaaa cctttttttt tatcatcatt attagcttac tttcataatt gcgactggtt
      841 ccaattgaca agcttttgat tttaacgact tttaacgaca acttgagaag atcaaaaaac
      901 aactaattat tcgaaacgag gaattcgagc tcggtacccg gggatccact agtctcgagc
      961 tgcaggcatg caagcttctt agacatgact gttcctcagt tcaagttggg cacttacgag
     1021 aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg cctgagagat
     1081 gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga ttttttttgt
     1141 cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag ctgatgaata
     1201 tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg tatttcccac
     1261 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1321 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1381 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1441 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1501 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1561 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1621 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1681 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1741 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     1801 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2941 tactcttcct ttttcaatga tctcctgatg actgactcac tgataataaa aatacggctt
     3001 cagaatttct caagactaca ctcactgtcc gacttcaagt atgacatttc ccttgctacc
     3061 tgcatacgca agtgttgcag agtttgataa ttccttgagt ttggtaggaa aagccgtgtt
     3121 tccctatgct gctgaccagc tgcacaacct gatcaagttc actcaatcga ctgagcttca
     3181 agttaatgtg caagttgagt catccgttac agaggaccaa tttgaggagc tgatcgacaa
     3241 cttgctcaag ttgtacaata atggtatcaa tgaagtgatt ttggacctag atttggcaga
     3301 aagagttgtc caaaggatga tcccaggcgc tagggttatc tataggaccc tggttgataa
     3361 agttgcatcc ttgcccgcta atgctagtat cgctgtgcct ttttcttctc cactgggcga
     3421 tttgaaaagt ttcactaatg gcggtagtag aactgtttat gctttttctg agaccgcaaa
     3481 gttggtagat gtgacttcca ctgttgcttc tggtataatc cccattattg atgctcggca
     3541 attgactact gaatacgaac tttctgaaga tgtcaaaaag ttccctgtca gtgaaatttt
     3601 gttggcgtct ttgactactg accgccccga tggtctattc actactttgg tggctgactc
     3661 ttctaattac tcgttgggcc tggtgtactc gtccaaaaag tctattccgg aggctataag
     3721 gacacaaact ggagtctacc aatctcgtcg tcacggtttg tggtataaag gtgctacatc
     3781 tggagcaact caaaagttgc tgggtatcga attggattgt gatggagact gcttgaaatt
     3841 tgtggttgaa caaacaggtg ttggtttctg tcacttggaa cgcacttcct gttttggcca
     3901 atcaaagggt cttagagcca tggaagccac cttgtgggat cgtaagagca atgctccaga
     3961 aggttcttat accaaacggt tatttgacga cgaagttttg ttgaacgcta aaattaggga
     4021 ggaagctgat gaacttgcag aagctaaatc caaggaagat atagcctggg aatgtgctga
     4081 cttattttat tttgcattag ttagatgtgc caagtacggt gtgacgttgg acgaggtgga
     4141 gagaaacctg gatatgaagt ccctaaaggt cactagaagg aaaggagatg ccaagccagg
     4201 atacaccaag gaacaaccta aagaagaatc caaacctaaa gaagtccctt ctgaaggtcg
     4261 tattgaattg tgcaaaattg acgtttctaa ggcctcctca caagaaattg aagatgccct
     4321 tcgtcgtcct atccagaaaa cggaacagat tatggaatta gtcaaaccaa ttgtcgacaa
     4381 tgttcgtcaa aatggtgaca aagccctttt agaactaact gccaagtttg atggagtcgc
     4441 tttgaagaca cctgtgttag aagctccttt cccagaggaa cttatgcaat tgccagataa
     4501 cgttaagaga gccattgatc tctctataga taacgtcagg aaattccatg aagctcaact
     4561 aacggagacg ttgcaagttg agacttgccc tggtgtagtc tgctctcgtt ttgcaagacc
     4621 tattgagaaa gttggcctct atattcctgg tggaaccgca attctgcctt ccacttccct
     4681 gatgctgggt gttcctgcca aagttgctgg tcgcaaagaa attgtttttg catctccacc
     4741 taagaaggat ggcaccctta ccccagaagt catctacgtt gcccacaagg ttggtgctaa
     4801 gtgtatcgtg ctagcaggag gcgcccaggc agtagctgct atggcttacg gaacagaaac
     4861 tgttcctaag tgtgacaaaa tatttggtcc aggaaaccag ttcgttactg ctgccaagat
     4921 gatggttcaa aatgacacat cagccctgtg tagtattgac atgcctgctg ggccttctga
     4981 agttctagtt attgctgata aatacgctga tccagatttc gttgcctcag accttctgtc
     5041 tcaagctgaa catggtattg attcccaggt gattctgttg gctgtcgata tgacagacaa
     5101 ggagcttgcc agaattgaag atgctgttca caaccaagct gtgcagttgc caagggttga
     5161 aattgtacgc aagtgtattg cacactctac aaccctatcg gttgcaacct acgagcaggc
     5221 tttggaaatg tccaatcagt acgctcctga acacttgatc ctgcaaatcg agaatgcttc
     5281 ttcttatgtt gatcaagtac aacacgctgg atctgtgttt gttggtgcct actctccaga
     5341 gagttgtgga gattactcct ccggcaccaa ccacactttg ccaacgtacg gatatgcccg
     5401 tcaatacagc ggagttaaca ctgcaacctt ccagaagttc atcacttcac aagacgtaac
     5461 tcctgaggga ctgaaacata ttggccaagc agtgatggat ctggctgctg ttgaaggtct
     5521 agatgctcac cgcaatgctg ttaaggttcg tatggagaaa ctgggactta tttaattatt
     5581 tagagatttt aacttacatt tagattcgat agatcattat tgaagcattt atcagggtta
     5641 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5701 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5761 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
