LOCUS       sequence-238923-        7812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-MET17 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 7812)
  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 7812)
  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-MET17
FEATURES             Location/Qualifiers
     source          1..7812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..50
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(88..110)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     210..229
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        420..795
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             796..2511
                     /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(2716..3171)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2853..2874)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3065..3084
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3297..3319
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3311..3328
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3312..3328
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3338..3357
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3338..3356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3381..3397
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3382..3398
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3432..3448)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3432..3448)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    3479..3982
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(4037..4057)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4037..4055)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4076..4092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4076..4092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4089..4111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4100..4116
                     /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(4124..4130,4131..4148,4149..4154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4169..4190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4307..4324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4478..5066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4558..4577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5237..6028,6029..6097))
                     /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     5860..5879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6098..6202)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(6253..7587)
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
ORIGIN
        1 gaggatggct atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
       61 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
      121 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
      181 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
      241 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
      301 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
      361 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagat ctgaattaat
      421 tcttgaataa tacataactt ttcttaaaag aatcaaagac agataaaatt taagagatat
      481 taaatattag tgagaagccg agaattttgt aacaccaaca taacactgac atctttaaca
      541 acttttaatt atgatacatt tcttacgtca tgattgatta ttacagctat gctgacaaat
      601 gactcttgtt gcatggctac gaaccgggta atactaagtg attgactctt gctgaccttt
      661 tattaagaac taaatggaca atattatgga gcatttcatg tataaattgg tgcgtaaaat
      721 cgttggatct ctcttctaag tacatcctac tataacaatc aagaaaaaca agaaaatcgg
      781 acaaaacaat caagtatgga ttctagaaca gttggtatat taggaggggg acaattggga
      841 cgtatgattg ttgaggcagc aaacaggctc aacattaaga cggtaatact agatgctgaa
      901 aattctcctg ccaaacaaat aagcaactcc aatgaccacg ttaatggctc cttttccaat
      961 cctcttgata tcgaaaaact agctgaaaaa tgtgatgtgc taacgattga gattgagcat
     1021 gttgatgttc ctacactaaa gaatcttcaa gtaaaacatc ccaaattaaa aatttaccct
     1081 tctccagaaa caatcagatt gatacaagac aaatatattc aaaaagagca tttaatcaaa
     1141 aatggtatag cagttaccca aagtgttcct gtggaacaag ccagtgagac gtccctattg
     1201 aatgttggaa gagatttggg ttttccattc gtcttgaagt cgaggacttt ggcatacgat
     1261 ggaagaggta acttcgttgt aaagaataag gaaatgattc cggaagcttt ggaagtactg
     1321 aaggatcgtc ctttgtacgc cgaaaaatgg gcaccattta ctaaagaatt agcagtcatg
     1381 attgtgaggt ctgttaacgg tttagtgttt tcttacccaa ttgtagagac tatccacaag
     1441 gacaatattt gtgacttatg ttatgcgcct gctagagttc cggactccgt tcaacttaag
     1501 gcgaagttgt tggcagaaaa tgcaatcaaa tcttttcccg gttgtggtat atttggtgtg
     1561 gaaatgttct atttagaaac aggggaattg cttattaacg aaattgcccc aaggcctcac
     1621 aactctggac attataccat tgatgcttgc gtcacttctc aatttgaagc tcatttgaga
     1681 tcaatattgg atttgccaat gccaaagaat ttcacatctt tctccaccat tacaacgaac
     1741 gccattatgc taaatgttct tggagacaaa catacaaaag ataaagagct agaaacttgc
     1801 gaaagagcat tggcgactcc aggttcctca gtgtacttat atggaaaaga gtctagacct
     1861 aacagaaaag taggtcacat aaatattatt gcctccagta tggcggaatg tgaacaaagg
     1921 ctgaactaca ttacaggtag aactgatatt ccaatcaaaa tctctgtcgc tcaaaagttg
     1981 gacttggaag caatggtcaa accattggtt ggaatcatca tgggatcaga ctctgacttg
     2041 ccggtaatgt ctgccgcatg tgcggtttta aaagattttg gcgttccatt tgaagtgaca
     2101 atagtctctg ctcatagaac tccacatagg atgtcagcat atgctatttc cgcaagcaag
     2161 cgtggaatta aaacaattat cgctggagct ggtggggctg ctcacttgcc aggtatggtg
     2221 gctgcaatga caccacttcc tgtcatcggt gtgcccgtaa aaggttcttg tctagatgga
     2281 gtagattctt tacattcaat tgtgcaaatg cctagaggtg ttccagtagc taccgtcgct
     2341 attaataata gtacgaacgc tgcgctgttg gctgtcagac tgcttggcgc ttatgattca
     2401 agttatacaa cgaaaatgga acagttttta ttaaagcaag aagaagaagt tcttgtcaaa
     2461 gcacaaaagt tagaaactgt cggttacgaa gcttatctag aaaacaagta atatataagt
     2521 ttattgatat acttgtacag caaataatta taaaatgata tacctatttt ttaggctttg
     2581 ttatgattac atcaaatgtg gacttcatac atagaaatca acgcttacag gtgtcctttt
     2641 ttaagaattt catacataag atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2701 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2761 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2821 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2881 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2941 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3001 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3061 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3121 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
     3181 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     3241 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     3301 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     3361 attgggtacc gggccccccc tcgaggtcga cggtatcgat aagcttgata tcgaattcct
     3421 gcagcccggg ggatccacta gttctagagc ggccgccctc ttagcgcttc ttaggacgga
     3481 tcgcttgcct gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt
     3541 tactctgtgt ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag
     3601 gtagaagagt tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa
     3661 aagcgtactt tacatatata tttattagac aagaaaagca gattaaatag atatacattc
     3721 gattaacgat aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac
     3781 aagatgaaac aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat
     3841 ttgttggcga tccccctaga gtcttttaca tcttcggaaa acaaaaacta ttttttcttt
     3901 aatttctttt tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat
     3961 aattattttt atagcacgtg atgaaaagga cccaggtggc gcggccgcca ccgcggtgga
     4021 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     4081 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4141 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4201 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4261 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4321 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4381 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4441 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4501 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4561 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4621 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4681 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4741 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4801 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4861 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4921 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4981 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5041 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5101 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5161 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5221 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5281 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5341 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5401 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5461 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5521 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5581 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5641 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5701 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5761 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5821 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5881 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5941 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6001 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6061 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6121 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6181 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctacaact
     6241 cattacgcac actcatggtt tttggccagc gaaaacagtt tcaaaagatt gctggaagtc
     6301 tgcaataatg tcatcaataa attcgatacc aacagagaca cgaattaagt ccttggtaac
     6361 accagatgcc aacttttctt tgtcatttaa ttgtttgtgg gtagtgaagt atggagcaat
     6421 gactaaggtc ttggcatcac caacattggc caagttagag gcaagcttta aattgtcaac
     6481 aacttgagca ccagaaagtt tgaatgggtc agtttccttg tcggcatttg gtaagtcttt
     6541 tacaccgaaa gataagacac caccgaaacc gttagataga tacttcttag cattttcatg
     6601 atgagaatga gatgctaaac cagggtatga aacccaagat acgtatgggg attgttctaa
     6661 ccatttggct aacttcaatg cattttcacc gtgtctttca gctctcaaag ataatgtttc
     6721 aacaccttgt agtagcaaga aagaggcaaa tgggttcatc aatggaccca aatctcttaa
     6781 tagttcagtt ctaacatgaa cgatgtatgc caagttaccg taggcttcat tgtagatagt
     6841 accgtgatat ccttcggcag gttgagagaa ttgagggaac ttttctgggt agtccttcca
     6901 tgggaactta ccagagtcaa caataatacc accgatagta gtaccatgac caccaatcca
     6961 tttggtagca gaatgtgtta caatatcagc accgtattta attggctgac agaagtaacc
     7021 accggcacca aatgtgttgt caacgacaac tggaataccg tgtttgtgag caattgcaac
     7081 aattttttca aaatccggaa cattgtactt tggattacca atggtttcca aataaacagc
     7141 cttggttctt tcatcaaaga ccttttcgaa ttcttctgga ttgtcacctt caacaaatct
     7201 agcctcgata ccaaatcttt tgaacgagat tttgaactgg ttataagtac caccgtataa
     7261 gtaagaagtg gaaacgatgt tgtcaccagt gtgtgccaaa ccttggatgg caagggtttg
     7321 agcggcttga ccggaggaaa cagccaaagc agcagcacca ccttctaaag cagcaattct
     7381 ttcttccaaa acattactgg ttgggttttg gaaacgggaa tagacgtaac ctggaacttc
     7441 tagaccaaac aattgcgaac catgcttaga gttttcgaaa acataagaag tggtggcgta
     7501 aattggtaca gctctggatc tgtgagcatt gtcaccaggg ttctcttggc cggcgtgtag
     7561 ttgaacagta tcgaaatgag atggcattgt atggatgggg gtaatagaat tgtatctatg
     7621 tatctgacga ccctgtatta cacgaaggaa atagtagatg aaaagacaag agagcaagaa
     7681 aaaggaaaaa cttcctttct aacagacgct ttacttaacc ttatatatat cttatttttt
     7741 tcatcgagcg tgttcaattg gacaaggtgc caccttttcg acacttcggt tattatgtta
     7801 cacagttttc at
//
