LOCUS       sequence-238924-        7314 bp    DNA     circular SYN 15-APR-2025
DEFINITION  ADE2-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 7314)
  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 7314)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 ADE2-URA3
FEATURES             Location/Qualifiers
     source          1..7314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     60..79
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        270..645
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             646..2361
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     rep_origin      complement(2566..3021)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2703..2724)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2915..2934
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3147..3169
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3161..3178
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3188..3207
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3231..3247
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3232..3248
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3282..3298)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3282..3298)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    3329..3832
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(3887..3907)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3887..3905)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3926..3942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3926..3942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3939..3961)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3950..3966
                     /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(3974..3980,3981..3998,3999..4004))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4019..4040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4157..4174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4328..4916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4408..4427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5087..5878,5879..5947))
                     /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     5710..5729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5948..6052)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(6138..6941)
                     /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(6942..7157)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     7196..7214
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7252..7274)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
       61 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
      121 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
      181 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
      241 tataaatcaa aagaatagat ctgaattaat tcttgaataa tacataactt ttcttaaaag
      301 aatcaaagac agataaaatt taagagatat taaatattag tgagaagccg agaattttgt
      361 aacaccaaca taacactgac atctttaaca acttttaatt atgatacatt tcttacgtca
      421 tgattgatta ttacagctat gctgacaaat gactcttgtt gcatggctac gaaccgggta
      481 atactaagtg attgactctt gctgaccttt tattaagaac taaatggaca atattatgga
      541 gcatttcatg tataaattgg tgcgtaaaat cgttggatct ctcttctaag tacatcctac
      601 tataacaatc aagaaaaaca agaaaatcgg acaaaacaat caagtatgga ttctagaaca
      661 gttggtatat taggaggggg acaattggga cgtatgattg ttgaggcagc aaacaggctc
      721 aacattaaga cggtaatact agatgctgaa aattctcctg ccaaacaaat aagcaactcc
      781 aatgaccacg ttaatggctc cttttccaat cctcttgata tcgaaaaact agctgaaaaa
      841 tgtgatgtgc taacgattga gattgagcat gttgatgttc ctacactaaa gaatcttcaa
      901 gtaaaacatc ccaaattaaa aatttaccct tctccagaaa caatcagatt gatacaagac
      961 aaatatattc aaaaagagca tttaatcaaa aatggtatag cagttaccca aagtgttcct
     1021 gtggaacaag ccagtgagac gtccctattg aatgttggaa gagatttggg ttttccattc
     1081 gtcttgaagt cgaggacttt ggcatacgat ggaagaggta acttcgttgt aaagaataag
     1141 gaaatgattc cggaagcttt ggaagtactg aaggatcgtc ctttgtacgc cgaaaaatgg
     1201 gcaccattta ctaaagaatt agcagtcatg attgtgaggt ctgttaacgg tttagtgttt
     1261 tcttacccaa ttgtagagac tatccacaag gacaatattt gtgacttatg ttatgcgcct
     1321 gctagagttc cggactccgt tcaacttaag gcgaagttgt tggcagaaaa tgcaatcaaa
     1381 tcttttcccg gttgtggtat atttggtgtg gaaatgttct atttagaaac aggggaattg
     1441 cttattaacg aaattgcccc aaggcctcac aactctggac attataccat tgatgcttgc
     1501 gtcacttctc aatttgaagc tcatttgaga tcaatattgg atttgccaat gccaaagaat
     1561 ttcacatctt tctccaccat tacaacgaac gccattatgc taaatgttct tggagacaaa
     1621 catacaaaag ataaagagct agaaacttgc gaaagagcat tggcgactcc aggttcctca
     1681 gtgtacttat atggaaaaga gtctagacct aacagaaaag taggtcacat aaatattatt
     1741 gcctccagta tggcggaatg tgaacaaagg ctgaactaca ttacaggtag aactgatatt
     1801 ccaatcaaaa tctctgtcgc tcaaaagttg gacttggaag caatggtcaa accattggtt
     1861 ggaatcatca tgggatcaga ctctgacttg ccggtaatgt ctgccgcatg tgcggtttta
     1921 aaagattttg gcgttccatt tgaagtgaca atagtctctg ctcatagaac tccacatagg
     1981 atgtcagcat atgctatttc cgcaagcaag cgtggaatta aaacaattat cgctggagct
     2041 ggtggggctg ctcacttgcc aggtatggtg gctgcaatga caccacttcc tgtcatcggt
     2101 gtgcccgtaa aaggttcttg tctagatgga gtagattctt tacattcaat tgtgcaaatg
     2161 cctagaggtg ttccagtagc taccgtcgct attaataata gtacgaacgc tgcgctgttg
     2221 gctgtcagac tgcttggcgc ttatgattca agttatacaa cgaaaatgga acagttttta
     2281 ttaaagcaag aagaagaagt tcttgtcaaa gcacaaaagt tagaaactgt cggttacgaa
     2341 gcttatctag aaaacaagta atatataagt ttattgatat acttgtacag caaataatta
     2401 taaaatgata tacctatttt ttaggctttg ttatgattac atcaaatgtg gacttcatac
     2461 atagaaatca acgcttacag gtgtcctttt ttaagaattt catacataag atctatgcgg
     2521 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta
     2581 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2641 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2701 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2761 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2821 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2881 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2941 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3001 atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3061 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3121 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3181 gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcga
     3241 cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta gttctagagc
     3301 ggccgccctc ttagcgcttc ttaggacgga tcgcttgcct gtaacttaca cgcgcctcgt
     3361 atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt ttatgttttg
     3421 tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga agaaaaaaaa
     3481 ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata tttattagac
     3541 aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg taaaatcaca
     3601 ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat taatacctga
     3661 gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga gtcttttaca
     3721 tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct atttttaatt
     3781 tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg atgaaaagga
     3841 cccaggtggc gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     3901 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3961 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4021 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4081 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4141 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4201 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4261 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4321 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4381 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4441 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4501 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4561 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4621 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4681 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4741 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4801 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4861 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4921 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4981 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5041 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5101 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5161 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5221 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5281 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5341 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5401 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5461 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5521 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5581 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5641 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5701 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5761 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5821 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5881 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5941 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6001 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6061 cccgaaaagt gccacctgac gtctgaagct ctaatttgtg agtttagtat acatgcattt
     6121 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
     6181 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
     6241 ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga
     6301 cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
     6361 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
     6421 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
     6481 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
     6541 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
     6601 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
     6661 aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
     6721 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
     6781 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
     6841 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
     6901 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
     6961 caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
     7021 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     7081 tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa agaataaaaa
     7141 aaaaatgatg aattgaaaag ctgtggtatg gtgcatcatg acattaacct ataaaaatag
     7201 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     7261 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcag
//
