LOCUS       sequence-238922-        8126 bp    DNA     circular SYN 15-APR-2025
DEFINITION  ADE2-LEU2 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 8126)
  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 8126)
  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-LEU2
FEATURES             Location/Qualifiers
     source          1..8126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..90
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(128..150)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     250..269
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        460..835
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             836..2551
                     /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(2756..3211)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2893..2914)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3105..3124
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3337..3359
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3351..3368
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3352..3368
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3378..3397
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3378..3396
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3421..3437
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3422..3438
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3472..3488)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3472..3488)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    3519..4022
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(4077..4097)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4077..4095)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4116..4132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4116..4132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4129..4151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4140..4156
                     /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(4164..4170,4171..4188,4189..4194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4209..4230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4347..4364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4518..5106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4598..4617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5277..6068,6069..6137))
                     /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     5900..5919
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6138..6242)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6410..6817
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6818..7912
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
        1 attgcacttt aacattaata ttgacaagga ggagggcacc acacaaaaag atcatgacat
       61 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
      121 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
      181 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
      241 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
      301 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
      361 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
      421 caaaatccct tataaatcaa aagaatagat ctgaattaat tcttgaataa tacataactt
      481 ttcttaaaag aatcaaagac agataaaatt taagagatat taaatattag tgagaagccg
      541 agaattttgt aacaccaaca taacactgac atctttaaca acttttaatt atgatacatt
      601 tcttacgtca tgattgatta ttacagctat gctgacaaat gactcttgtt gcatggctac
      661 gaaccgggta atactaagtg attgactctt gctgaccttt tattaagaac taaatggaca
      721 atattatgga gcatttcatg tataaattgg tgcgtaaaat cgttggatct ctcttctaag
      781 tacatcctac tataacaatc aagaaaaaca agaaaatcgg acaaaacaat caagtatgga
      841 ttctagaaca gttggtatat taggaggggg acaattggga cgtatgattg ttgaggcagc
      901 aaacaggctc aacattaaga cggtaatact agatgctgaa aattctcctg ccaaacaaat
      961 aagcaactcc aatgaccacg ttaatggctc cttttccaat cctcttgata tcgaaaaact
     1021 agctgaaaaa tgtgatgtgc taacgattga gattgagcat gttgatgttc ctacactaaa
     1081 gaatcttcaa gtaaaacatc ccaaattaaa aatttaccct tctccagaaa caatcagatt
     1141 gatacaagac aaatatattc aaaaagagca tttaatcaaa aatggtatag cagttaccca
     1201 aagtgttcct gtggaacaag ccagtgagac gtccctattg aatgttggaa gagatttggg
     1261 ttttccattc gtcttgaagt cgaggacttt ggcatacgat ggaagaggta acttcgttgt
     1321 aaagaataag gaaatgattc cggaagcttt ggaagtactg aaggatcgtc ctttgtacgc
     1381 cgaaaaatgg gcaccattta ctaaagaatt agcagtcatg attgtgaggt ctgttaacgg
     1441 tttagtgttt tcttacccaa ttgtagagac tatccacaag gacaatattt gtgacttatg
     1501 ttatgcgcct gctagagttc cggactccgt tcaacttaag gcgaagttgt tggcagaaaa
     1561 tgcaatcaaa tcttttcccg gttgtggtat atttggtgtg gaaatgttct atttagaaac
     1621 aggggaattg cttattaacg aaattgcccc aaggcctcac aactctggac attataccat
     1681 tgatgcttgc gtcacttctc aatttgaagc tcatttgaga tcaatattgg atttgccaat
     1741 gccaaagaat ttcacatctt tctccaccat tacaacgaac gccattatgc taaatgttct
     1801 tggagacaaa catacaaaag ataaagagct agaaacttgc gaaagagcat tggcgactcc
     1861 aggttcctca gtgtacttat atggaaaaga gtctagacct aacagaaaag taggtcacat
     1921 aaatattatt gcctccagta tggcggaatg tgaacaaagg ctgaactaca ttacaggtag
     1981 aactgatatt ccaatcaaaa tctctgtcgc tcaaaagttg gacttggaag caatggtcaa
     2041 accattggtt ggaatcatca tgggatcaga ctctgacttg ccggtaatgt ctgccgcatg
     2101 tgcggtttta aaagattttg gcgttccatt tgaagtgaca atagtctctg ctcatagaac
     2161 tccacatagg atgtcagcat atgctatttc cgcaagcaag cgtggaatta aaacaattat
     2221 cgctggagct ggtggggctg ctcacttgcc aggtatggtg gctgcaatga caccacttcc
     2281 tgtcatcggt gtgcccgtaa aaggttcttg tctagatgga gtagattctt tacattcaat
     2341 tgtgcaaatg cctagaggtg ttccagtagc taccgtcgct attaataata gtacgaacgc
     2401 tgcgctgttg gctgtcagac tgcttggcgc ttatgattca agttatacaa cgaaaatgga
     2461 acagttttta ttaaagcaag aagaagaagt tcttgtcaaa gcacaaaagt tagaaactgt
     2521 cggttacgaa gcttatctag aaaacaagta atatataagt ttattgatat acttgtacag
     2581 caaataatta taaaatgata tacctatttt ttaggctttg ttatgattac atcaaatgtg
     2641 gacttcatac atagaaatca acgcttacag gtgtcctttt ttaagaattt catacataag
     2701 atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     2761 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2821 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2881 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2941 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     3001 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     3061 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     3121 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     3181 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     3241 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     3301 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     3361 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc
     3421 tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta
     3481 gttctagagc ggccgccctc ttagcgcttc ttaggacgga tcgcttgcct gtaacttaca
     3541 cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt
     3601 ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga
     3661 agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata
     3721 tttattagac aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg
     3781 taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat
     3841 taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga
     3901 gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct
     3961 atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg
     4021 atgaaaagga cccaggtggc gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     4081 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4141 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4201 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4261 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4321 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4381 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4441 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4501 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4561 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4621 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4681 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4741 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4801 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4861 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4921 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4981 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5041 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5101 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5161 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5221 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5281 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5341 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5401 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5461 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5521 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5581 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5641 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5701 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5761 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5821 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5881 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5941 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6001 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6061 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6121 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6181 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6241 cgcacatttc cccgaaaagt gccacctgac gtctcctgta cttccttgtt catgtgtgtt
     6301 caaaaacgtt atatttatag gataattata ctctatttct caacaagtaa ttggttgttt
     6361 ggccgagcgg tctaaggcgc ctgattcaag aaatatcttg accgcagtta actgtgggaa
     6421 tactcaggta tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc
     6481 aacataacga gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt
     6541 ttttgttaat ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga
     6601 aaaaggaaag gtgagagcgc cggaaccggc ttttcatata gaatagagaa gcgttcatga
     6661 ctaaatgctt gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt
     6721 tccaataggt ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc
     6781 aatatatata tatatatttc aaggatatac cattctaatg tctgccccta agaagatcgt
     6841 cgttttgcca ggtgaccacg ttggtcaaga aatcacagcc gaagccatta aggttcttaa
     6901 agctatttct gatgttcgtt ccaatgtcaa gttcgatttc gaaaatcatt taattggtgg
     6961 tgctgctatc gatgctacag gtgttccact tccagatgag gcgctggaag cctccaagaa
     7021 ggctgatgcc gttttgttag gtgctgtggg tggtcctaaa tggggtaccg gtagtgttag
     7081 acctgaacaa ggtttactaa aaatccgtaa agaacttcaa ttgtacgcca acttaagacc
     7141 atgtaacttt gcatccgact ctcttttaga cttatctcca atcaagccac aatttgctaa
     7201 aggtactgac ttcgttgttg tcagagaatt agtgggaggt atttactttg gtaagagaaa
     7261 ggaagacgat ggtgatggtg tcgcttggga tagtgaacaa tacaccgttc cagaagtgca
     7321 aagaatcaca agaatggccg ctttcatggc cctacaacat gagccaccat tgcctatttg
     7381 gtccttggat aaagctaatg ttttggcctc ttcaagatta tggagaaaaa ctgtggagga
     7441 aaccatcaag aacgaattcc ctacattgaa ggttcaacat caattgattg attctgccgc
     7501 catgatccta gttaagaacc caacccacct aaatggtatt ataatcacca gcaacatgtt
     7561 tggtgatatc atctccgatg aagcctccgt tatcccaggt tccttgggtt tgttgccatc
     7621 tgcgtccttg gcctctttgc cagacaagaa caccgcattt ggtttgtacg aaccatgcca
     7681 cggttctgct ccagatttgc caaagaataa ggtcaaccct atcgccacta tcttgtctgc
     7741 tgcaatgatg ttgaaattgt cattgaactt gcctgaagaa ggtaaggcca ttgaagatgc
     7801 agttaaaaag gttttggatg caggtatcag aactggtgat ttaggtggtt ccaacagtac
     7861 caccgaagtc ggtgatgctg tcgccgaaga agttaagaaa atccttgctt aaaaagattc
     7921 tcttttttta tgatatttgt acataaactt tataaatgaa attcataata gaaacgacac
     7981 gaaattacaa aatggaatat gttcataggg tagacgaaac tatatacgca atctacatac
     8041 atttatcaag aaggagaaaa aggaggatgt aaaggaatac aggtaagcaa attgatacta
     8101 atggctcaac gtgataagga aaaaga
//
