LOCUS       sequence-238921-        7274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-HIS3 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 7274)
  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 7274)
  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 ADE2-HIS3
FEATURES             Location/Qualifiers
     source          1..7274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..30
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(68..90)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     190..209
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        400..775
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             776..2491
                     /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(2696..3151)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2833..2854)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3045..3064
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3277..3299
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3291..3308
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3292..3308
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3318..3337
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3318..3336
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3361..3377
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3362..3378
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3412..3428)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3412..3428)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    3459..3962
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(4017..4037)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4017..4035)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4056..4072)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4056..4072)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4069..4091)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4080..4096
                     /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(4104..4110,4111..4128,4129..4134))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4149..4170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4287..4304)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4458..5046)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4538..4557)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5217..6008,6009..6077))
                     /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     5840..5859
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6078..6182)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6264..6451
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             6452..7114
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
ORIGIN
        1 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
       61 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
      121 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
      181 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
      241 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
      301 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
      361 caaaatccct tataaatcaa aagaatagat ctgaattaat tcttgaataa tacataactt
      421 ttcttaaaag aatcaaagac agataaaatt taagagatat taaatattag tgagaagccg
      481 agaattttgt aacaccaaca taacactgac atctttaaca acttttaatt atgatacatt
      541 tcttacgtca tgattgatta ttacagctat gctgacaaat gactcttgtt gcatggctac
      601 gaaccgggta atactaagtg attgactctt gctgaccttt tattaagaac taaatggaca
      661 atattatgga gcatttcatg tataaattgg tgcgtaaaat cgttggatct ctcttctaag
      721 tacatcctac tataacaatc aagaaaaaca agaaaatcgg acaaaacaat caagtatgga
      781 ttctagaaca gttggtatat taggaggggg acaattggga cgtatgattg ttgaggcagc
      841 aaacaggctc aacattaaga cggtaatact agatgctgaa aattctcctg ccaaacaaat
      901 aagcaactcc aatgaccacg ttaatggctc cttttccaat cctcttgata tcgaaaaact
      961 agctgaaaaa tgtgatgtgc taacgattga gattgagcat gttgatgttc ctacactaaa
     1021 gaatcttcaa gtaaaacatc ccaaattaaa aatttaccct tctccagaaa caatcagatt
     1081 gatacaagac aaatatattc aaaaagagca tttaatcaaa aatggtatag cagttaccca
     1141 aagtgttcct gtggaacaag ccagtgagac gtccctattg aatgttggaa gagatttggg
     1201 ttttccattc gtcttgaagt cgaggacttt ggcatacgat ggaagaggta acttcgttgt
     1261 aaagaataag gaaatgattc cggaagcttt ggaagtactg aaggatcgtc ctttgtacgc
     1321 cgaaaaatgg gcaccattta ctaaagaatt agcagtcatg attgtgaggt ctgttaacgg
     1381 tttagtgttt tcttacccaa ttgtagagac tatccacaag gacaatattt gtgacttatg
     1441 ttatgcgcct gctagagttc cggactccgt tcaacttaag gcgaagttgt tggcagaaaa
     1501 tgcaatcaaa tcttttcccg gttgtggtat atttggtgtg gaaatgttct atttagaaac
     1561 aggggaattg cttattaacg aaattgcccc aaggcctcac aactctggac attataccat
     1621 tgatgcttgc gtcacttctc aatttgaagc tcatttgaga tcaatattgg atttgccaat
     1681 gccaaagaat ttcacatctt tctccaccat tacaacgaac gccattatgc taaatgttct
     1741 tggagacaaa catacaaaag ataaagagct agaaacttgc gaaagagcat tggcgactcc
     1801 aggttcctca gtgtacttat atggaaaaga gtctagacct aacagaaaag taggtcacat
     1861 aaatattatt gcctccagta tggcggaatg tgaacaaagg ctgaactaca ttacaggtag
     1921 aactgatatt ccaatcaaaa tctctgtcgc tcaaaagttg gacttggaag caatggtcaa
     1981 accattggtt ggaatcatca tgggatcaga ctctgacttg ccggtaatgt ctgccgcatg
     2041 tgcggtttta aaagattttg gcgttccatt tgaagtgaca atagtctctg ctcatagaac
     2101 tccacatagg atgtcagcat atgctatttc cgcaagcaag cgtggaatta aaacaattat
     2161 cgctggagct ggtggggctg ctcacttgcc aggtatggtg gctgcaatga caccacttcc
     2221 tgtcatcggt gtgcccgtaa aaggttcttg tctagatgga gtagattctt tacattcaat
     2281 tgtgcaaatg cctagaggtg ttccagtagc taccgtcgct attaataata gtacgaacgc
     2341 tgcgctgttg gctgtcagac tgcttggcgc ttatgattca agttatacaa cgaaaatgga
     2401 acagttttta ttaaagcaag aagaagaagt tcttgtcaaa gcacaaaagt tagaaactgt
     2461 cggttacgaa gcttatctag aaaacaagta atatataagt ttattgatat acttgtacag
     2521 caaataatta taaaatgata tacctatttt ttaggctttg ttatgattac atcaaatgtg
     2581 gacttcatac atagaaatca acgcttacag gtgtcctttt ttaagaattt catacataag
     2641 atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     2701 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2761 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2821 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2881 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2941 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3001 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3061 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3121 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     3181 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     3241 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     3301 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc
     3361 tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta
     3421 gttctagagc ggccgccctc ttagcgcttc ttaggacgga tcgcttgcct gtaacttaca
     3481 cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt
     3541 ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga
     3601 agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata
     3661 tttattagac aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg
     3721 taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat
     3781 taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga
     3841 gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct
     3901 atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg
     3961 atgaaaagga cccaggtggc gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     4021 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4081 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4141 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4201 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4261 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4321 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4381 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4441 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4501 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4561 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4621 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4681 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4741 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4801 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4861 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4921 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4981 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5041 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5101 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5161 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5221 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5281 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5341 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5401 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5461 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5521 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5581 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5641 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5701 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5761 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5821 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5881 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5941 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6001 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6061 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6121 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6181 cgcacatttc cccgaaaagt gccacctgac gtctaacaca gtcctttccc gcaattttct
     6241 ttttctatta ctcttggcct cctctagtac actctatatt tttttatgcc tcggtaatga
     6301 ttttcatttt tttttttcca cctagcggat gactcttttt ttttcttagc gattggcatt
     6361 atcacataat gaattataca ttatataaag taatgtgatt tcttcgaaga atatactaaa
     6421 aaatgagcag gcaagataaa cgaaggcaaa gatgacagag cagaaagccc tagtaaagcg
     6481 tattacaaat gaaaccaaga ttcagattgc gatctcttta aagggtggtc ccctagcgat
     6541 agagcactcg atcttcccag aaaaagaggc agaagcagta gcagaacagg ccacacaatc
     6601 gcaagtgatt aacgtccaca caggtatagg gtttctggac catatgatac atgctctggc
     6661 caagcattcc ggctggtcgc taatcgttga gtgcattggt gacttacaca tagacgacca
     6721 tcacaccact gaagactgcg ggattgctct cggtcaagct tttaaagagg ccctaggggc
     6781 cgtgcgtgga gtaaaaaggt ttggatcagg atttgcgcct ttggatgagg cactttccag
     6841 agcggtggta gatctttcga acaggccgta cgcagttgtc gaacttggtt tgcaaaggga
     6901 gaaagtagga gatctctctt gcgagatgat cccgcatttt cttgaaagct ttgcagaggc
     6961 tagcagaatt accctccacg ttgattgtct gcgaggcaag aatgatcatc accgtagtga
     7021 gagtgcgttc aaggctcttg cggttgccat aagagaagcc acctcgccca atggtaccaa
     7081 cgatgttccc tccaccaaag gtgttcttat gtagtgacac cgattattta aagctgcagc
     7141 atacgatata tatacatgtg tatatatgta tacctatgaa tgtcagtaag tatgtatacg
     7201 aacagtatga tactgaagat gacaaggtaa tgcatcattc tatacgtgtc attctgaacg
     7261 aggcatcatg acat
//
