LOCUS       sequence-234438-        7713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-HphMx 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 7713)
  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 7713)
  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-HphMx
FEATURES             Location/Qualifiers
     source          1..7713
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    11..514
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(569..589)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(569..587)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(608..624)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(608..624)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(621..643)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    632..648
                     /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(656..662,663..680,681..686))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(839..856)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1010..1598)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1090..1109)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1769..2560,2561..2629))
                     /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     2392..2411
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2630..2734)
                     /gene="bla"
                     /label=AmpR promoter
     gene            2807..4382
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        2807..3150
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             3151..4179
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             3151..4179
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      4185..4382
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     4435..4453
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4491..4513)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4613..4632
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4661..5036
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             5037..6752
                     /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(6957..7412)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7094..7115)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7306..7325
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7542..7564
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7556..7573
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7557..7573
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7583..7602
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7583..7601
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7626..7642
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7627..7643
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7677..7693)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7677..7693)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 tcttaggacg gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat
       61 aatttgggaa tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag
      121 taaataaaga aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa
      181 aatttcaaca aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat
      241 agatatacat tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc
      301 ttctacacag acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat
      361 aaaaggtagt atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac
      421 tattttttct ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa
      481 aatttaaatt ataattattt ttatagcacg tgatgaaaag gacccaggtg gcgcggccgc
      541 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
      601 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      661 taggagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagg taactcacat
      721 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      781 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      841 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      901 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      961 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1021 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1081 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1141 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1201 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1261 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1321 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1381 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1441 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1501 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1561 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1621 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1681 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1741 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1801 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1861 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1921 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1981 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2041 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2101 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2161 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2221 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2281 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2341 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2401 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2461 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2521 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2581 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2701 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2761 acgtcttgtt tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca
     2821 gaataccctc cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc
     2881 gtacatttag cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg
     2941 cacggcgcga agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc
     3001 cctcacagac gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta
     3061 ggatttgcca ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag
     3121 ttctcacatc acatccgaac ataaacaacc atgggtaaaa agcctgaact caccgcgacg
     3181 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     3241 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     3301 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     3361 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     3421 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     3481 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     3541 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     3601 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     3661 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     3721 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     3781 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     3841 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     3901 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     3961 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     4021 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     4081 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     4141 aaccgacgcc ccagcactcg tccgagggca aaggaataat cagtactgac aataaaaaga
     4201 ttcttgtttt caagaacttg tcatttgtat agttttttta tattgtagtt gttctatttt
     4261 aatcaaatgt tagcgtgatt tatatttttt ttcgcctcga catcatctgc ccagatgcga
     4321 agttaagtgc gcagaaagta atatcatgcg tcaatcgtat gtgaatgctg gtcgctatac
     4381 tgctgtcgat tcgatactaa cgccgccatc cagatcatga cattaaccta taaaaatagg
     4441 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4501 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4561 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4621 gagcagattg tactgagagt gcaccataga tctgaattaa ttcttgaata atacataact
     4681 tttcttaaaa gaatcaaaga cagataaaat ttaagagata ttaaatatta gtgagaagcc
     4741 gagaattttg taacaccaac ataacactga catctttaac aacttttaat tatgatacat
     4801 ttcttacgtc atgattgatt attacagcta tgctgacaaa tgactcttgt tgcatggcta
     4861 cgaaccgggt aatactaagt gattgactct tgctgacctt ttattaagaa ctaaatggac
     4921 aatattatgg agcatttcat gtataaattg gtgcgtaaaa tcgttggatc tctcttctaa
     4981 gtacatccta ctataacaat caagaaaaac aagaaaatcg gacaaaacaa tcaagtatgg
     5041 attctagaac agttggtata ttaggagggg gacaattggg acgtatgatt gttgaggcag
     5101 caaacaggct caacattaag acggtaatac tagatgctga aaattctcct gccaaacaaa
     5161 taagcaactc caatgaccac gttaatggct ccttttccaa tcctcttgat atcgaaaaac
     5221 tagctgaaaa atgtgatgtg ctaacgattg agattgagca tgttgatgtt cctacactaa
     5281 agaatcttca agtaaaacat cccaaattaa aaatttaccc ttctccagaa acaatcagat
     5341 tgatacaaga caaatatatt caaaaagagc atttaatcaa aaatggtata gcagttaccc
     5401 aaagtgttcc tgtggaacaa gccagtgaga cgtccctatt gaatgttgga agagatttgg
     5461 gttttccatt cgtcttgaag tcgaggactt tggcatacga tggaagaggt aacttcgttg
     5521 taaagaataa ggaaatgatt ccggaagctt tggaagtact gaaggatcgt cctttgtacg
     5581 ccgaaaaatg ggcaccattt actaaagaat tagcagtcat gattgtgagg tctgttaacg
     5641 gtttagtgtt ttcttaccca attgtagaga ctatccacaa ggacaatatt tgtgacttat
     5701 gttatgcgcc tgctagagtt ccggactccg ttcaacttaa ggcgaagttg ttggcagaaa
     5761 atgcaatcaa atcttttccc ggttgtggta tatttggtgt ggaaatgttc tatttagaaa
     5821 caggggaatt gcttattaac gaaattgccc caaggcctca caactctgga cattatacca
     5881 ttgatgcttg cgtcacttct caatttgaag ctcatttgag atcaatattg gatttgccaa
     5941 tgccaaagaa tttcacatct ttctccacca ttacaacgaa cgccattatg ctaaatgttc
     6001 ttggagacaa acatacaaaa gataaagagc tagaaacttg cgaaagagca ttggcgactc
     6061 caggttcctc agtgtactta tatggaaaag agtctagacc taacagaaaa gtaggtcaca
     6121 taaatattat tgcctccagt atggcggaat gtgaacaaag gctgaactac attacaggta
     6181 gaactgatat tccaatcaaa atctctgtcg ctcaaaagtt ggacttggaa gcaatggtca
     6241 aaccattggt tggaatcatc atgggatcag actctgactt gccggtaatg tctgccgcat
     6301 gtgcggtttt aaaagatttt ggcgttccat ttgaagtgac aatagtctct gctcatagaa
     6361 ctccacatag gatgtcagca tatgctattt ccgcaagcaa gcgtggaatt aaaacaatta
     6421 tcgctggagc tggtggggct gctcacttgc caggtatggt ggctgcaatg acaccacttc
     6481 ctgtcatcgg tgtgcccgta aaaggttctt gtctagatgg agtagattct ttacattcaa
     6541 ttgtgcaaat gcctagaggt gttccagtag ctaccgtcgc tattaataat agtacgaacg
     6601 ctgcgctgtt ggctgtcaga ctgcttggcg cttatgattc aagttataca acgaaaatgg
     6661 aacagttttt attaaagcaa gaagaagaag ttcttgtcaa agcacaaaag ttagaaactg
     6721 tcggttacga agcttatcta gaaaacaagt aatatataag tttattgata tacttgtaca
     6781 gcaaataatt ataaaatgat atacctattt tttaggcttt gttatgatta catcaaatgt
     6841 ggacttcata catagaaatc aacgcttaca ggtgtccttt tttaagaatt tcatacataa
     6901 gatctatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     6961 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     7021 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7081 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     7141 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     7201 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     7261 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     7321 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     7381 cgccgcgctt aatgcgccgc tacagggcgc gtcgcgccat tcgccattca ggctgcgcaa
     7441 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     7501 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     7561 aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc
     7621 ccccctcgag gtcgacggta tcgataagct tgatatcgaa ttcctgcagc ccgggggatc
     7681 cactagttct agagcggccg ccctcttagc gct
//
