LOCUS       sequence-43435-e        5568 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 5568)
  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 5568)
  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: pIB3
FEATURES             Location/Qualifiers
     source          1..5568
                     /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(1092..1680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1172..1191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1851..2642,2643..2711))
                     /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     2474..2493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             2811..5345
                     /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     5541..5559
                     /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 accatatgat gagtcacaat ctgcttccac agacgagtac aaggacaggc aaaaggaatt
      241 ggaagaagtt gctaacccaa taatgagcaa gttctatgga gctgctggtg gagctcctgg
      301 tggagctcct ggtggcttcc ctggaggttt ccctggcgga gctggcgcag ctggcggtgc
      361 cccaggtggt gctgccccag gcggagacag cggaccaacc gtggaagaag tcgattaagc
      421 aattcaacgg ataaattctg gttaatatat ataacgtgaa taggaaatta aggaaatttt
      481 ggatctaata atgtgctgta tgccgacatc gggcatcgta gattgtatag tatcgctgac
      541 actataataa gccagccaaa acccctaaac cagttgccct ccactaatta gtgtactacc
      601 caatcttgcc tcttcgggtg tcttttataa ggacagattc acaagctctt gttgcccaat
      661 acacacatac acacagagat aatagcagtc gaattcgagc tcggtacccg gggatccact
      721 agtctcgagc tgcaggcatg caagcttctt agacatgact gttcctcagt tcaagttggg
      781 cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg
      841 cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga
      901 ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag
      961 ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg
     1021 tatttcccac acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     1081 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     1141 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     1201 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     1261 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     1321 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     1381 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     1441 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     1501 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     1561 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     1621 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     1681 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     1741 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     1801 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     1861 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1921 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1981 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     2041 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     2101 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     2161 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     2221 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     2281 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     2341 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     2401 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     2461 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2521 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2581 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2641 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2701 tgaatactca tactcttcct ttttcaatga tctcctgatg actgactcac tgataataaa
     2761 aatacggctt cagaatttct caagactaca ctcactgtcc gacttcaagt atgacatttc
     2821 ccttgctacc tgcatacgca agtgttgcag agtttgataa ttccttgagt ttggtaggaa
     2881 aagccgtgtt tccctatgct gctgaccagc tgcacaacct gatcaagttc actcaatcga
     2941 ctgagcttca agttaatgtg caagttgagt catccgttac agaggaccaa tttgaggagc
     3001 tgatcgacaa cttgctcaag ttgtacaata atggtatcaa tgaagtgatt ttggacctag
     3061 atttggcaga aagagttgtc caaaggatga tcccaggcgc tagggttatc tataggaccc
     3121 tggttgataa agttgcatcc ttgcccgcta atgctagtat cgctgtgcct ttttcttctc
     3181 cactgggcga tttgaaaagt ttcactaatg gcggtagtag aactgtttat gctttttctg
     3241 agaccgcaaa gttggtagat gtgacttcca ctgttgcttc tggtataatc cccattattg
     3301 atgctcggca attgactact gaatacgaac tttctgaaga tgtcaaaaag ttccctgtca
     3361 gtgaaatttt gttggcgtct ttgactactg accgccccga tggtctattc actactttgg
     3421 tggctgactc ttctaattac tcgttgggcc tggtgtactc gtccaaaaag tctattccgg
     3481 aggctataag gacacaaact ggagtctacc aatctcgtcg tcacggtttg tggtataaag
     3541 gtgctacatc tggagcaact caaaagttgc tgggtatcga attggattgt gatggagact
     3601 gcttgaaatt tgtggttgaa caaacaggtg ttggtttctg tcacttggaa cgcacttcct
     3661 gttttggcca atcaaagggt cttagagcca tggaagccac cttgtgggat cgtaagagca
     3721 atgctccaga aggttcttat accaaacggt tatttgacga cgaagttttg ttgaacgcta
     3781 aaattaggga ggaagctgat gaacttgcag aagctaaatc caaggaagat atagcctggg
     3841 aatgtgctga cttattttat tttgcattag ttagatgtgc caagtacggt gtgacgttgg
     3901 acgaggtgga gagaaacctg gatatgaagt ccctaaaggt cactagaagg aaaggagatg
     3961 ccaagccagg atacaccaag gaacaaccta aagaagaatc caaacctaaa gaagtccctt
     4021 ctgaaggtcg tattgaattg tgcaaaattg acgtttctaa ggcctcctca caagaaattg
     4081 aagatgccct tcgtcgtcct atccagaaaa cggaacagat tatggaatta gtcaaaccaa
     4141 ttgtcgacaa tgttcgtcaa aatggtgaca aagccctttt agaactaact gccaagtttg
     4201 atggagtcgc tttgaagaca cctgtgttag aagctccttt cccagaggaa cttatgcaat
     4261 tgccagataa cgttaagaga gccattgatc tctctataga taacgtcagg aaattccatg
     4321 aagctcaact aacggagacg ttgcaagttg agacttgccc tggtgtagtc tgctctcgtt
     4381 ttgcaagacc tattgagaaa gttggcctct atattcctgg tggaaccgca attctgcctt
     4441 ccacttccct gatgctgggt gttcctgcca aagttgctgg tcgcaaagaa attgtttttg
     4501 catctccacc taagaaggat ggcaccctta ccccagaagt catctacgtt gcccacaagg
     4561 ttggtgctaa gtgtatcgtg ctagcaggag gcgcccaggc agtagctgct atggcttacg
     4621 gaacagaaac tgttcctaag tgtgacaaaa tatttggtcc aggaaaccag ttcgttactg
     4681 ctgccaagat gatggttcaa aatgacacat cagccctgtg tagtattgac atgcctgctg
     4741 ggccttctga agttctagtt attgctgata aatacgctga tccagatttc gttgcctcag
     4801 accttctgtc tcaagctgaa catggtattg attcccaggt gattctgttg gctgtcgata
     4861 tgacagacaa ggagcttgcc agaattgaag atgctgttca caaccaagct gtgcagttgc
     4921 caagggttga aattgtacgc aagtgtattg cacactctac aaccctatcg gttgcaacct
     4981 acgagcaggc tttggaaatg tccaatcagt acgctcctga acacttgatc ctgcaaatcg
     5041 agaatgcttc ttcttatgtt gatcaagtac aacacgctgg atctgtgttt gttggtgcct
     5101 actctccaga gagttgtgga gattactcct ccggcaccaa ccacactttg ccaacgtacg
     5161 gatatgcccg tcaatacagc ggagttaaca ctgcaacctt ccagaagttc atcacttcac
     5221 aagacgtaac tcctgaggga ctgaaacata ttggccaagc agtgatggat ctggctgctg
     5281 ttgaaggtct agatgctcac cgcaatgctg ttaaggttcg tatggagaaa ctgggactta
     5341 tttaattatt tagagatttt aacttacatt tagattcgat agatcattat tgaagcattt
     5401 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5461 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     5521 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
