LOCUS       sequence-238930-        5924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-URA3 ICE plasmid used to facilitate yeast recombination cloning
            and stable genomic Integration after CEN Excision..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5924)
  AUTHORS   Flagg MP, Kao A, Hampton RY
  TITLE     ICE plasmids: combining the ease of yeast recombination cloning with
            the stability of genomic integration.
  JOURNAL   Yeast. 2019 May 10. doi: 10.1002/yea.3400.
  PUBMED    31074531
REFERENCE   2  (bases 1 to 5924)
  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. 2019
            May 10. doi: 10.1002/yea.3400."
COMMENT     Sequence Label: pICE TRP1-URA3
FEATURES             Location/Qualifiers
     source          1..5924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(7..29)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     129..148
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        172..387
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             388..1191
                     /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"
     rep_origin      complement(1322..1777)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1459..1480)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1671..1690
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1903..1925
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1917..1934
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1918..1934
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1944..1963
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1944..1962
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1987..2003
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1988..2004
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2038..2054)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2038..2054)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    2085..2588
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(2643..2663)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2643..2661)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2682..2698)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2682..2698)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2695..2717)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2706..2722
                     /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)."
     promoter        complement(join(2730..2736,2737..2754,2755..2760))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2775..2796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2913..2930)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3084..3672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3164..3183)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3843..4634,4635..4703))
                     /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     4466..4485
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4704..4808)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4849..5130
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5131..5805
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     5875..5893
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
       61 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
      121 taactatgcg gcatcagagc agattgtact gagagtgcac cataccacct tttcaattca
      181 tcattttttt tttattcttt tttttgattt cggtttcctt gaaatttttt tgattcggta
      241 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
      301 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
      361 aaaacctgca ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct
      421 actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac
      481 ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta
      541 ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag
      601 ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaagac
      661 agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga
      721 atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc
      781 ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca
      841 gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt
      901 gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga
      961 gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagac
     1021 gcattgggtc aacagtatag aaccgtggat gatgtggtct ctacaggatc tgacattatt
     1081 attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     1141 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     1201 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     1261 ttattaccct atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     1321 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     1381 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     1441 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     1501 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     1561 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     1621 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     1681 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     1741 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca
     1801 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     1861 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     1921 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccgggc
     1981 cccccctcga ggtcgacggt atcgataagc ttgatatcga attcctgcag cccgggggat
     2041 ccactagttc tagagcggcc gccctcttag cgcttcttag gacggatcgc ttgcctgtaa
     2101 cttacacgcg cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat
     2161 ttatttttat gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg
     2221 gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca
     2281 tatatattta ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt
     2341 aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt
     2401 cggcattaat acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc
     2461 cctagagtct tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta
     2521 ctttctattt ttaatttata tatttatatt aaaaaattta aattataatt atttttatag
     2581 cacgtgatga aaaggaccca ggtggcgcgg ccgccaccgc ggtggagctc cagcttttgt
     2641 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     2701 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2761 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2821 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2881 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2941 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3001 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3061 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3121 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3181 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3241 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3301 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3361 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3421 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3481 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3541 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3601 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3661 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3721 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3781 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3841 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3901 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3961 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4021 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4081 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4141 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4201 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4261 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4321 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4381 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4441 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4501 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4561 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4621 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4681 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4741 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4801 gttccgcgca catttccccg aaaagtgcca cctgacgtct gcaccataaa cgacattact
     4861 atatatataa tataggaagc atttaataga acagcatcgt aatatatgtg tactttgcag
     4921 ttatgacgcc agatggcagt agtggaagat attctttatt gaaaaatagc ttgtcacctt
     4981 acgtacaatc ttgatccgga gcttttcttt ttttgccgat taagaattaa ttcggtcgaa
     5041 aaaagaaaag gagagggcca agagggaggg cattggtgac tattgagcac gtgagtatac
     5101 gtgattaagc acacaaaggc agcttggagt atgtctgtta ttaatttcac aggtagttct
     5161 ggtccattgg tgaaagtttg cggcttgcag agcacagagg ccgcagaatg tgctctagat
     5221 tccgatgctg acttgctggg tattatatgt gtgcccaata gaaagagaac aattgacccg
     5281 gttattgcaa ggaaaatttc aagtcttgta aaagcatata aaaatagttc aggcactccg
     5341 aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg atgttttggc tctggtcaat
     5401 gattacggca ttgatatcgt ccaactgcat ggagatgagt cgtggcaaga ataccaagag
     5461 ttcctcggtt tgccagttat taaaagactc gtatttccaa aagactgcaa catactactc
     5521 agtgcagctt cacagaaacc tcattcgttt attcccttgt ttgattcaga agcaggtggg
     5581 acaggtgaac ttttggattg gaactcgatt tctgactggg ttggaaggca agagagcccc
     5641 gaaagcttac attttatgtt agctggtgga ctgacgccag aaaatgttgg tgatgcgctt
     5701 agattaaatg gcgttattgg tgttgatgta agcggaggtg tggagacaaa tggtgtaaaa
     5761 gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga aataggttat tactgagtag
     5821 tatttattta agtattgttt gtgcacttgc ctaatcatga cattaaccta taaaaatagg
     5881 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acgg
//
