LOCUS       sequence-234407-        6080 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 6080)
  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 6080)
  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..6080
                     /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
     primer_bind     2802..2820
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2858..2880)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2980..2999
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3028..3403
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             3404..5119
                     /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(5324..5779)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5461..5482)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5673..5692
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5909..5931
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5923..5940
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5924..5940
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5950..5969
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5950..5968
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5993..6009
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5994..6010
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6044..6060)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6044..6060)
                     /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 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2821 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2881 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2941 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     3001 tgagagtgca ccatagatct gaattaattc ttgaataata cataactttt cttaaaagaa
     3061 tcaaagacag ataaaattta agagatatta aatattagtg agaagccgag aattttgtaa
     3121 caccaacata acactgacat ctttaacaac ttttaattat gatacatttc ttacgtcatg
     3181 attgattatt acagctatgc tgacaaatga ctcttgttgc atggctacga accgggtaat
     3241 actaagtgat tgactcttgc tgacctttta ttaagaacta aatggacaat attatggagc
     3301 atttcatgta taaattggtg cgtaaaatcg ttggatctct cttctaagta catcctacta
     3361 taacaatcaa gaaaaacaag aaaatcggac aaaacaatca agtatggatt ctagaacagt
     3421 tggtatatta ggagggggac aattgggacg tatgattgtt gaggcagcaa acaggctcaa
     3481 cattaagacg gtaatactag atgctgaaaa ttctcctgcc aaacaaataa gcaactccaa
     3541 tgaccacgtt aatggctcct tttccaatcc tcttgatatc gaaaaactag ctgaaaaatg
     3601 tgatgtgcta acgattgaga ttgagcatgt tgatgttcct acactaaaga atcttcaagt
     3661 aaaacatccc aaattaaaaa tttacccttc tccagaaaca atcagattga tacaagacaa
     3721 atatattcaa aaagagcatt taatcaaaaa tggtatagca gttacccaaa gtgttcctgt
     3781 ggaacaagcc agtgagacgt ccctattgaa tgttggaaga gatttgggtt ttccattcgt
     3841 cttgaagtcg aggactttgg catacgatgg aagaggtaac ttcgttgtaa agaataagga
     3901 aatgattccg gaagctttgg aagtactgaa ggatcgtcct ttgtacgccg aaaaatgggc
     3961 accatttact aaagaattag cagtcatgat tgtgaggtct gttaacggtt tagtgttttc
     4021 ttacccaatt gtagagacta tccacaagga caatatttgt gacttatgtt atgcgcctgc
     4081 tagagttccg gactccgttc aacttaaggc gaagttgttg gcagaaaatg caatcaaatc
     4141 ttttcccggt tgtggtatat ttggtgtgga aatgttctat ttagaaacag gggaattgct
     4201 tattaacgaa attgccccaa ggcctcacaa ctctggacat tataccattg atgcttgcgt
     4261 cacttctcaa tttgaagctc atttgagatc aatattggat ttgccaatgc caaagaattt
     4321 cacatctttc tccaccatta caacgaacgc cattatgcta aatgttcttg gagacaaaca
     4381 tacaaaagat aaagagctag aaacttgcga aagagcattg gcgactccag gttcctcagt
     4441 gtacttatat ggaaaagagt ctagacctaa cagaaaagta ggtcacataa atattattgc
     4501 ctccagtatg gcggaatgtg aacaaaggct gaactacatt acaggtagaa ctgatattcc
     4561 aatcaaaatc tctgtcgctc aaaagttgga cttggaagca atggtcaaac cattggttgg
     4621 aatcatcatg ggatcagact ctgacttgcc ggtaatgtct gccgcatgtg cggttttaaa
     4681 agattttggc gttccatttg aagtgacaat agtctctgct catagaactc cacataggat
     4741 gtcagcatat gctatttccg caagcaagcg tggaattaaa acaattatcg ctggagctgg
     4801 tggggctgct cacttgccag gtatggtggc tgcaatgaca ccacttcctg tcatcggtgt
     4861 gcccgtaaaa ggttcttgtc tagatggagt agattcttta cattcaattg tgcaaatgcc
     4921 tagaggtgtt ccagtagcta ccgtcgctat taataatagt acgaacgctg cgctgttggc
     4981 tgtcagactg cttggcgctt atgattcaag ttatacaacg aaaatggaac agtttttatt
     5041 aaagcaagaa gaagaagttc ttgtcaaagc acaaaagtta gaaactgtcg gttacgaagc
     5101 ttatctagaa aacaagtaat atataagttt attgatatac ttgtacagca aataattata
     5161 aaatgatata cctatttttt aggctttgtt atgattacat caaatgtgga cttcatacat
     5221 agaaatcaac gcttacaggt gtcctttttt aagaatttca tacataagat ctatgcggtg
     5281 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aaacgttaat
     5341 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5401 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5461 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5521 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5581 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5641 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5701 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5761 gcgccgctac agggcgcgtc gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5821 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     5881 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     5941 gagcgcgcgt aatacgactc actatagggc gaattgggta ccgggccccc cctcgaggtc
     6001 gacggtatcg ataagcttga tatcgaattc ctgcagcccg ggggatccac tagttctaga
     6061 gcggccgccc tcttagcgct
//
