LOCUS       sequence-239159-        7652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-KanMx 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 7652)
  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 7652)
  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-KanMx
FEATURES             Location/Qualifiers
     source          1..7652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..117
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        308..683
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             684..2399
                     /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(2604..3059)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2741..2762)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2953..2972
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3185..3207
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3199..3216
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3200..3216
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3226..3245
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3226..3244
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3269..3285
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3270..3286
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3320..3336)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3320..3336)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    3367..3870
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(3925..3945)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3925..3943)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3964..3980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3964..3980)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3977..3999)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3988..4004
                     /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(4012..4018,4019..4036,4037..4042))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4057..4078
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4195..4212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4366..4954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4446..4465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5125..5916,5917..5985))
                     /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     5748..5767
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5986..6090)
                     /gene="bla"
                     /label=AmpR promoter
     gene            6163..7519
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        6163..6506
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             6507..7316
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(6574..6593)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      7322..7519
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     7572..7590
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7628..7650)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
       61 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      121 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      181 aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
      241 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagatct
      301 gaattaattc ttgaataata cataactttt cttaaaagaa tcaaagacag ataaaattta
      361 agagatatta aatattagtg agaagccgag aattttgtaa caccaacata acactgacat
      421 ctttaacaac ttttaattat gatacatttc ttacgtcatg attgattatt acagctatgc
      481 tgacaaatga ctcttgttgc atggctacga accgggtaat actaagtgat tgactcttgc
      541 tgacctttta ttaagaacta aatggacaat attatggagc atttcatgta taaattggtg
      601 cgtaaaatcg ttggatctct cttctaagta catcctacta taacaatcaa gaaaaacaag
      661 aaaatcggac aaaacaatca agtatggatt ctagaacagt tggtatatta ggagggggac
      721 aattgggacg tatgattgtt gaggcagcaa acaggctcaa cattaagacg gtaatactag
      781 atgctgaaaa ttctcctgcc aaacaaataa gcaactccaa tgaccacgtt aatggctcct
      841 tttccaatcc tcttgatatc gaaaaactag ctgaaaaatg tgatgtgcta acgattgaga
      901 ttgagcatgt tgatgttcct acactaaaga atcttcaagt aaaacatccc aaattaaaaa
      961 tttacccttc tccagaaaca atcagattga tacaagacaa atatattcaa aaagagcatt
     1021 taatcaaaaa tggtatagca gttacccaaa gtgttcctgt ggaacaagcc agtgagacgt
     1081 ccctattgaa tgttggaaga gatttgggtt ttccattcgt cttgaagtcg aggactttgg
     1141 catacgatgg aagaggtaac ttcgttgtaa agaataagga aatgattccg gaagctttgg
     1201 aagtactgaa ggatcgtcct ttgtacgccg aaaaatgggc accatttact aaagaattag
     1261 cagtcatgat tgtgaggtct gttaacggtt tagtgttttc ttacccaatt gtagagacta
     1321 tccacaagga caatatttgt gacttatgtt atgcgcctgc tagagttccg gactccgttc
     1381 aacttaaggc gaagttgttg gcagaaaatg caatcaaatc ttttcccggt tgtggtatat
     1441 ttggtgtgga aatgttctat ttagaaacag gggaattgct tattaacgaa attgccccaa
     1501 ggcctcacaa ctctggacat tataccattg atgcttgcgt cacttctcaa tttgaagctc
     1561 atttgagatc aatattggat ttgccaatgc caaagaattt cacatctttc tccaccatta
     1621 caacgaacgc cattatgcta aatgttcttg gagacaaaca tacaaaagat aaagagctag
     1681 aaacttgcga aagagcattg gcgactccag gttcctcagt gtacttatat ggaaaagagt
     1741 ctagacctaa cagaaaagta ggtcacataa atattattgc ctccagtatg gcggaatgtg
     1801 aacaaaggct gaactacatt acaggtagaa ctgatattcc aatcaaaatc tctgtcgctc
     1861 aaaagttgga cttggaagca atggtcaaac cattggttgg aatcatcatg ggatcagact
     1921 ctgacttgcc ggtaatgtct gccgcatgtg cggttttaaa agattttggc gttccatttg
     1981 aagtgacaat agtctctgct catagaactc cacataggat gtcagcatat gctatttccg
     2041 caagcaagcg tggaattaaa acaattatcg ctggagctgg tggggctgct cacttgccag
     2101 gtatggtggc tgcaatgaca ccacttcctg tcatcggtgt gcccgtaaaa ggttcttgtc
     2161 tagatggagt agattcttta cattcaattg tgcaaatgcc tagaggtgtt ccagtagcta
     2221 ccgtcgctat taataatagt acgaacgctg cgctgttggc tgtcagactg cttggcgctt
     2281 atgattcaag ttatacaacg aaaatggaac agtttttatt aaagcaagaa gaagaagttc
     2341 ttgtcaaagc acaaaagtta gaaactgtcg gttacgaagc ttatctagaa aacaagtaat
     2401 atataagttt attgatatac ttgtacagca aataattata aaatgatata cctatttttt
     2461 aggctttgtt atgattacat caaatgtgga cttcatacat agaaatcaac gcttacaggt
     2521 gtcctttttt aagaatttca tacataagat ctatgcggtg tgaaataccg cacagatgcg
     2581 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     2641 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2701 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2761 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2821 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2881 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2941 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     3001 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     3061 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     3121 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     3181 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     3241 tagggcgaat tgggtaccgg gccccccctc gaggtcgacg gtatcgataa gcttgatatc
     3301 gaattcctgc agcccggggg atccactagt tctagagcgg ccgccctctt agcgcttctt
     3361 aggacggatc gcttgcctgt aacttacacg cgcctcgtat cttttaatga tggaataatt
     3421 tgggaattta ctctgtgttt atttattttt atgttttgta tttggatttt agaaagtaaa
     3481 taaagaaggt agaagagtta cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt
     3541 tcaacaaaaa gcgtacttta catatatatt tattagacaa gaaaagcaga ttaaatagat
     3601 atacattcga ttaacgataa gtaaaatgta aaatcacagg attttcgtgt gtggtcttct
     3661 acacagacaa gatgaaacaa ttcggcatta atacctgaga gcaggaagag caagataaaa
     3721 ggtagtattt gttggcgatc cccctagagt cttttacatc ttcggaaaac aaaaactatt
     3781 ttttctttaa tttctttttt tactttctat ttttaattta tatatttata ttaaaaaatt
     3841 taaattataa ttatttttat agcacgtgat gaaaaggacc caggtggcgc ggccgccacc
     3901 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     3961 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4021 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4081 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4141 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4201 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4261 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4321 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4381 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4441 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4501 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4561 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4621 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4681 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4741 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4801 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4861 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4921 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4981 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5041 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5101 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5161 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5221 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5281 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5341 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5401 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5461 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5521 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5581 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5641 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5701 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5761 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5821 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5881 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5941 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6001 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6061 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6121 cttgtttagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat
     6181 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
     6241 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
     6301 gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc
     6361 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
     6421 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
     6481 cacatcacat ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc
     6541 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     6601 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     6661 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     6721 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     6781 catggttact caccactgcg atccccggca aaacagcatt ccaggtatta gaagaatatc
     6841 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     6901 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     6961 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     7021 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     7081 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     7141 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     7201 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     7261 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     7321 tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat
     7381 tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat
     7441 catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg
     7501 aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag atcatgacat
     7561 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
     7621 gtgaaaacct ctgacacatg cagctcccgg ag
//
