LOCUS       sequence-238615-        6238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2 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 6238)
  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 6238)
  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
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        126..501
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             502..2217
                     /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(2422..2877)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2559..2580)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2771..2790
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3003..3025
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3017..3034
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3018..3034
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3044..3063
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3044..3062
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3087..3103
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3088..3104
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3138..3154)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3138..3154)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    3185..3688
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(3743..3763)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3743..3761)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3782..3798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3782..3798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3795..3817)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3806..3822
                     /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(3830..3836,3837..3854,3855..3860))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3875..3896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4013..4030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4184..4772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4264..4283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4943..5734,5735..5803))
                     /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     5566..5585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5804..5908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5976..5994
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6032..6054)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6154..6173
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
       61 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagatctga
      121 attaattctt gaataataca taacttttct taaaagaatc aaagacagat aaaatttaag
      181 agatattaaa tattagtgag aagccgagaa ttttgtaaca ccaacataac actgacatct
      241 ttaacaactt ttaattatga tacatttctt acgtcatgat tgattattac agctatgctg
      301 acaaatgact cttgttgcat ggctacgaac cgggtaatac taagtgattg actcttgctg
      361 accttttatt aagaactaaa tggacaatat tatggagcat ttcatgtata aattggtgcg
      421 taaaatcgtt ggatctctct tctaagtaca tcctactata acaatcaaga aaaacaagaa
      481 aatcggacaa aacaatcaag tatggattct agaacagttg gtatattagg agggggacaa
      541 ttgggacgta tgattgttga ggcagcaaac aggctcaaca ttaagacggt aatactagat
      601 gctgaaaatt ctcctgccaa acaaataagc aactccaatg accacgttaa tggctccttt
      661 tccaatcctc ttgatatcga aaaactagct gaaaaatgtg atgtgctaac gattgagatt
      721 gagcatgttg atgttcctac actaaagaat cttcaagtaa aacatcccaa attaaaaatt
      781 tacccttctc cagaaacaat cagattgata caagacaaat atattcaaaa agagcattta
      841 atcaaaaatg gtatagcagt tacccaaagt gttcctgtgg aacaagccag tgagacgtcc
      901 ctattgaatg ttggaagaga tttgggtttt ccattcgtct tgaagtcgag gactttggca
      961 tacgatggaa gaggtaactt cgttgtaaag aataaggaaa tgattccgga agctttggaa
     1021 gtactgaagg atcgtccttt gtacgccgaa aaatgggcac catttactaa agaattagca
     1081 gtcatgattg tgaggtctgt taacggttta gtgttttctt acccaattgt agagactatc
     1141 cacaaggaca atatttgtga cttatgttat gcgcctgcta gagttccgga ctccgttcaa
     1201 cttaaggcga agttgttggc agaaaatgca atcaaatctt ttcccggttg tggtatattt
     1261 ggtgtggaaa tgttctattt agaaacaggg gaattgctta ttaacgaaat tgccccaagg
     1321 cctcacaact ctggacatta taccattgat gcttgcgtca cttctcaatt tgaagctcat
     1381 ttgagatcaa tattggattt gccaatgcca aagaatttca catctttctc caccattaca
     1441 acgaacgcca ttatgctaaa tgttcttgga gacaaacata caaaagataa agagctagaa
     1501 acttgcgaaa gagcattggc gactccaggt tcctcagtgt acttatatgg aaaagagtct
     1561 agacctaaca gaaaagtagg tcacataaat attattgcct ccagtatggc ggaatgtgaa
     1621 caaaggctga actacattac aggtagaact gatattccaa tcaaaatctc tgtcgctcaa
     1681 aagttggact tggaagcaat ggtcaaacca ttggttggaa tcatcatggg atcagactct
     1741 gacttgccgg taatgtctgc cgcatgtgcg gttttaaaag attttggcgt tccatttgaa
     1801 gtgacaatag tctctgctca tagaactcca cataggatgt cagcatatgc tatttccgca
     1861 agcaagcgtg gaattaaaac aattatcgct ggagctggtg gggctgctca cttgccaggt
     1921 atggtggctg caatgacacc acttcctgtc atcggtgtgc ccgtaaaagg ttcttgtcta
     1981 gatggagtag attctttaca ttcaattgtg caaatgccta gaggtgttcc agtagctacc
     2041 gtcgctatta ataatagtac gaacgctgcg ctgttggctg tcagactgct tggcgcttat
     2101 gattcaagtt atacaacgaa aatggaacag tttttattaa agcaagaaga agaagttctt
     2161 gtcaaagcac aaaagttaga aactgtcggt tacgaagctt atctagaaaa caagtaatat
     2221 ataagtttat tgatatactt gtacagcaaa taattataaa atgatatacc tattttttag
     2281 gctttgttat gattacatca aatgtggact tcatacatag aaatcaacgc ttacaggtgt
     2341 ccttttttaa gaatttcata cataagatct atgcggtgtg aaataccgca cagatgcgta
     2401 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2461 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2521 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2581 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2641 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2701 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2761 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2821 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     2881 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     2941 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     3001 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     3061 gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga
     3121 attcctgcag cccgggggat ccactagttc tagagcggcc gccctcttag cgcttcttag
     3181 gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg gaataatttg
     3241 ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag aaagtaaata
     3301 aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt aaaaaatttc
     3361 aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt aaatagatat
     3421 acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt ggtcttctac
     3481 acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca agataaaagg
     3541 tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa aaactatttt
     3601 ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt aaaaaattta
     3661 aattataatt atttttatag cacgtgatga aaaggaccca ggtggcgcgg ccgccaccgc
     3721 ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     3781 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3841 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3901 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3961 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4021 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4081 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4141 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4201 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4261 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4321 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4381 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4441 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4501 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4561 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4621 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4681 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4741 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4801 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4861 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4921 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4981 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5041 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5101 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5161 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5221 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5281 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5341 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5401 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5461 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5521 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5581 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5641 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5701 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5761 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5821 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5881 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5941 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     6001 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     6061 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6121 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6181 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcag
//
