LOCUS       sequence-43433-e        5273 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 5273)
  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 5273)
  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: pIB1
FEATURES             Location/Qualifiers
     source          1..5273
                     /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"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(797..1385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(877..896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1556..2347,2348..2416))
                     /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     2179..2198
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             2516..5050
                     /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     5246..5264
                     /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 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggat
      421 ccactagtct cgagctgcag gcatgcaagc ttcttagaca tgactgttcc tcagttcaag
      481 ttgggcactt acgagaagac cggtcttgct agattctaat caagaggatg tcagaatgcc
      541 atttgcctga gagatgcagg cttcattttt gatacttttt tatttgtaac ctatatagta
      601 taggattttt tttgtcattt tgtttcttct cgtacgagct tgctcctgat cagcctatct
      661 cgcagctgat gaatatcttg tggtaggggt ttgggaaaat cattcgagtt tgatgttttt
      721 cttggtattt cccacacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      781 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      841 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      901 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      961 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1021 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1081 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1141 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1201 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1261 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1321 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1381 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1441 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1501 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1561 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1621 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1681 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1741 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1801 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1861 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1921 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1981 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2041 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     2101 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     2161 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     2221 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     2281 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2341 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2401 aatgttgaat actcatactc ttcctttttc aatgatctcc tgatgactga ctcactgata
     2461 ataaaaatac ggcttcagaa tttctcaaga ctacactcac tgtccgactt caagtatgac
     2521 atttcccttg ctacctgcat acgcaagtgt tgcagagttt gataattcct tgagtttggt
     2581 aggaaaagcc gtgtttccct atgctgctga ccagctgcac aacctgatca agttcactca
     2641 atcgactgag cttcaagtta atgtgcaagt tgagtcatcc gttacagagg accaatttga
     2701 ggagctgatc gacaacttgc tcaagttgta caataatggt atcaatgaag tgattttgga
     2761 cctagatttg gcagaaagag ttgtccaaag gatgatccca ggcgctaggg ttatctatag
     2821 gaccctggtt gataaagttg catccttgcc cgctaatgct agtatcgctg tgcctttttc
     2881 ttctccactg ggcgatttga aaagtttcac taatggcggt agtagaactg tttatgcttt
     2941 ttctgagacc gcaaagttgg tagatgtgac ttccactgtt gcttctggta taatccccat
     3001 tattgatgct cggcaattga ctactgaata cgaactttct gaagatgtca aaaagttccc
     3061 tgtcagtgaa attttgttgg cgtctttgac tactgaccgc cccgatggtc tattcactac
     3121 tttggtggct gactcttcta attactcgtt gggcctggtg tactcgtcca aaaagtctat
     3181 tccggaggct ataaggacac aaactggagt ctaccaatct cgtcgtcacg gtttgtggta
     3241 taaaggtgct acatctggag caactcaaaa gttgctgggt atcgaattgg attgtgatgg
     3301 agactgcttg aaatttgtgg ttgaacaaac aggtgttggt ttctgtcact tggaacgcac
     3361 ttcctgtttt ggccaatcaa agggtcttag agccatggaa gccaccttgt gggatcgtaa
     3421 gagcaatgct ccagaaggtt cttataccaa acggttattt gacgacgaag ttttgttgaa
     3481 cgctaaaatt agggaggaag ctgatgaact tgcagaagct aaatccaagg aagatatagc
     3541 ctgggaatgt gctgacttat tttattttgc attagttaga tgtgccaagt acggtgtgac
     3601 gttggacgag gtggagagaa acctggatat gaagtcccta aaggtcacta gaaggaaagg
     3661 agatgccaag ccaggataca ccaaggaaca acctaaagaa gaatccaaac ctaaagaagt
     3721 cccttctgaa ggtcgtattg aattgtgcaa aattgacgtt tctaaggcct cctcacaaga
     3781 aattgaagat gcccttcgtc gtcctatcca gaaaacggaa cagattatgg aattagtcaa
     3841 accaattgtc gacaatgttc gtcaaaatgg tgacaaagcc cttttagaac taactgccaa
     3901 gtttgatgga gtcgctttga agacacctgt gttagaagct cctttcccag aggaacttat
     3961 gcaattgcca gataacgtta agagagccat tgatctctct atagataacg tcaggaaatt
     4021 ccatgaagct caactaacgg agacgttgca agttgagact tgccctggtg tagtctgctc
     4081 tcgttttgca agacctattg agaaagttgg cctctatatt cctggtggaa ccgcaattct
     4141 gccttccact tccctgatgc tgggtgttcc tgccaaagtt gctggtcgca aagaaattgt
     4201 ttttgcatct ccacctaaga aggatggcac ccttacccca gaagtcatct acgttgccca
     4261 caaggttggt gctaagtgta tcgtgctagc aggaggcgcc caggcagtag ctgctatggc
     4321 ttacggaaca gaaactgttc ctaagtgtga caaaatattt ggtccaggaa accagttcgt
     4381 tactgctgcc aagatgatgg ttcaaaatga cacatcagcc ctgtgtagta ttgacatgcc
     4441 tgctgggcct tctgaagttc tagttattgc tgataaatac gctgatccag atttcgttgc
     4501 ctcagacctt ctgtctcaag ctgaacatgg tattgattcc caggtgattc tgttggctgt
     4561 cgatatgaca gacaaggagc ttgccagaat tgaagatgct gttcacaacc aagctgtgca
     4621 gttgccaagg gttgaaattg tacgcaagtg tattgcacac tctacaaccc tatcggttgc
     4681 aacctacgag caggctttgg aaatgtccaa tcagtacgct cctgaacact tgatcctgca
     4741 aatcgagaat gcttcttctt atgttgatca agtacaacac gctggatctg tgtttgttgg
     4801 tgcctactct ccagagagtt gtggagatta ctcctccggc accaaccaca ctttgccaac
     4861 gtacggatat gcccgtcaat acagcggagt taacactgca accttccaga agttcatcac
     4921 ttcacaagac gtaactcctg agggactgaa acatattggc caagcagtga tggatctggc
     4981 tgctgttgaa ggtctagatg ctcaccgcaa tgctgttaag gttcgtatgg agaaactggg
     5041 acttatttaa ttatttagag attttaactt acatttagat tcgatagatc attattgaag
     5101 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5161 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     5221 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtc
//
