LOCUS       sequence-345484-        7060 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 7060)
  AUTHORS   Boder ET, Wittrup KD
  TITLE     Yeast surface display for screening combinatorial polypeptide 
            libraries.
  JOURNAL   Nat Biotechnol. 1997 Jun;15(6):553-7.
  PUBMED    9181578
REFERENCE   2  (bases 1 to 7060)
  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: "Nat 
            Biotechnol."; date: "1997-06"; volume: "15(6)"; pages: "553-7"
COMMENT     Sequence Label: pIU211
FEATURES             Location/Qualifiers
     source          1..7060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..665
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    217..334
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     629..652
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(3211..3228)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3211..3227)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3220..3242)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3436..3455)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3555..3577
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3695..4498)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(4499..4714)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        4865..4969
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4970..5038,5039..5830)
                     /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     complement(5188..5207)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6001..6589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6490..6509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6743..6760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6877..6898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6913..6918,6919..6936,6937..6943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6951..6967
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6956..6978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6975..6991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6975..6991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tttcaaaaat tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca
       61 tggcattacc accatataca tatccatata catatccata tctaatctta cttatatgtt
      121 gtggaaatgt aaagagcccc attatcttag cctaaaaaaa ccttctcttt ggaactttca
      181 gtaatacgct taactgctca ttgctatatt gaagtacgga ttagaagccg ccgagcgggt
      241 gacagccctc cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct
      301 gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc
      361 ttttatggtt atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaatgaa
      421 cgaatcaaat taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg
      481 gtaattaatc agcgaagcga tgatttttga tctattaaca gatatataaa tgcaaaaact
      541 gcataaccac tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat
      601 gtaataaaag tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa
      661 aaaaccccgg atcgaattca gctaaaaaaa ccaaaaaatt tcagatgaca ttatctttcg
      721 ctcattttac ctacctgttc acaatattgt tgggattaac taatattgcc ttggcatctg
      781 atccagaaac gattctagtg acgataacca agacaaacga tgcaaatggg gttgttacaa
      841 ctacagtttc acccgcgcta gtctccacat ccactatcgt tcaagctggc actacgacat
      901 tgtatacgac ttggtgtcca ttgacggtat ccacttcatc tgctgccgaa ataagtcctt
      961 caatatcgta cgctactacc ctatccagat ttagtacttt gacattatct acagaagtct
     1021 gctcccatga ggcatgtcct tcgtcatcga cgttgccaac caccacctta tctgtgactt
     1081 ccaagttcac ttcatatatt tgccctactt gtcacacaac cgctatcagc tcattatccg
     1141 aagtaggaac tacaaccgtg gtatcatcca gcgccattga accatcaagt gcctctataa
     1201 tctcacctgt cacctctaca ctttcgagta caacatcgtc caatccaact actacctccc
     1261 taagttcgac atctacatct ccaagctcta catctacatc tccaagctct acatctacct
     1321 catcaagttc gacatctacc tcatcaagtt cgacatctac ctcatcaagt tcgacatcta
     1381 catctccaag ttcgacatcc acatcttcaa gtttgacatc cacatcttca agttctacat
     1441 ctacatccca aagttctaca tctacctcat caagttcgac atctacatct ccaagctcta
     1501 catctacctc atcaagttca acatctacat ctccaagttc taaatctact tctgcaagct
     1561 ccacttccac ttcttcatat tcaacatcta catccccaag tttgacttct tcatctccaa
     1621 ctttggcttc cacttctcca agttcaacat ctattagctc tacttttact gattcaactt
     1681 catcccttgg ctcctctata gcatcttcat caacgtctgt gtcattatac agcccatcca
     1741 cacctgttta ctccgtccct tcgacttcgt caaatgttgc aactccttct atgacttctt
     1801 caactgttga aacaactgtt agttcacaaa gttcgtctga atatatcacc aaatcctcaa
     1861 tttctactac tatcccatca ttttccatgt ctacatattt caccactgtt agtggagtca
     1921 ctacaatgta tacgacatgg tgtccttata gctctgaatc tgagactagc acattaacca
     1981 gtatgcatga aacggttaca acagacgcta cagtctgcac tcacgagtct tgcatgccct
     2041 cgcagacaac aagtttgatt acatcttcta taaaaatgtc cactaaaaac gtcgcaactt
     2101 ctgtaagcac ctcaacggtt gaatcctcat atgcatgctc cacatgtgct gaaacgtcac
     2161 actcgtattc ttccgtgcaa acagcttcat caagttctgt aacacagcag accacatcca
     2221 caaagagttg ggtaagttca atgacaactt cggatgaaga tttcaataag cacgctaccg
     2281 gtaagtatca tgtaacatct tcaggtacct caaccatttc gactagtgta agtgaagcca
     2341 cgagtacatc aagcattgac tcagaatctc aagaacaatc atcacactta ttatcgacat
     2401 cggtcctttc atcctcctcc ttgtctgcta cattatcctc tgacagtact attttgctat
     2461 tcagttctgt atcatcacta agtgtcgaac agtcaccagt taccacactt caaatttctt
     2521 caacatcaga gattttacaa cccacttctt ccacagctat tgctacaata tctgcctcta
     2581 catcatcact ttccgcaaca tctatctcta caccatctac ctctgtggaa tcgactattg
     2641 aatcttcatc attgactccg acggtatctt ctattttcct ctcatcatca tctgctccct
     2701 cttctctaca aacatctgtt accactacag aagtttccac tacttcaatc tccatacaat
     2761 accaaacttc atcaatggta acaattagcc aatatatggg cagtggatcg caaacgcgtt
     2821 tgccattagg aaagttggtc ttcgccatca tggcagttgc ctgcaatgta attttcagtt
     2881 aatagctcga gatctgataa caacagtgta gatgtaacaa aatcgacttt gttcccactg
     2941 tacttttagc tcgtacaaaa tacaatatac ttttcatttc tccgtaaaca acatgttttc
     3001 ccatgtaata tccttttcta tttttcgttc cgttaccaac tttacacata ctttatatag
     3061 ctattcactt ctatacacta aaaaactaag acaattttaa ttttgctgcc tgccatattt
     3121 caatttgtta taaattccta taatttatcc tattagtagc taaaaaaaga tgaatgtgaa
     3181 tcgaatccta agagaattga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     3241 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3301 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3361 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3421 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     3481 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3541 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3601 cgcgagacga aagggcgggt aataactgat ataattaaat tgaagctcta atttgtgagt
     3661 ttagtataca tgcatttact tataatacag ttttttagtt ttgctggccg catcttctca
     3721 aatatgcttc ccagcctgct tttctgtaac gttcaccctc taccttagca tcccttccct
     3781 ttgcaaatag tcctcttcca acaataataa tgtcagatcc tgtagagacc acatcatcca
     3841 cggttctata ctgttgaccc aatgcgtctc ccttgtcatc taaacccaca ccgggtgtca
     3901 taatcaacca atcgtaacct tcatctcttc cacccatgtc tctttgagca ataaagccga
     3961 taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc cttagtatat tctccagtag
     4021 atagggagcc cttgcatgac aattctgcta acatcaaaag gcctctaggt tcctttgtta
     4081 cttcttctgc cgcctgcttc aaaccgctaa caatacctgg gcccaccaca ccgtgtgcat
     4141 tcgtaatgtc tgcccattct gctattctgt atacacccgc agagtactgc aatttgactg
     4201 tattaccaat gtcagcaaat tttctgtctt cgaagagtaa aaaattgtac ttggcggata
     4261 atgcctttag cggcttaact gtgccctcca tggaaaaatc agtcaagata tccacatgtg
     4321 tttttagtaa acaaattttg ggacctaatg cttcaactaa ctccagtaat tccttggtgg
     4381 tacgaacatc caatgaagca cacaagtttg tttgcttttc gtgcatgata ttaaatagct
     4441 tggcagcaac aggactagga tgagtagcag cacgttcctt atatgtagct ttcgacatga
     4501 tttatcttcg tttcctgcat gtttttgttc tgtgcagttg ggttaagaat actgggcaat
     4561 ttcatgtttc ttcaacacta catatgcgta tatataccaa tctaagtctg tgctccttcc
     4621 ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa atttcaagga aaccgaaatc
     4681 aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaagctg gcctcgtgat acgcctattt
     4741 ttataggtta atgtcatgat aataacctat aaaaataggc gtatcacgag gcctattttt
     4801 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4861 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4921 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4981 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5041 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5101 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5161 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5221 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5281 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5341 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     5401 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5461 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5521 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5581 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5641 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5701 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5761 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5821 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5881 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5941 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6001 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6061 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     6781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     6841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     6901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     6961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca
     7021 ggtcgactct agaggatccc cgggtaccaa ttccttgaat
//
