LOCUS       sequence-238939-        6762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-URA3 integrating (YIp) vector for stable genomic integration in
            the BY4741 background..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6762)
  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 6762)
  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: YIp ADE2-URA3
FEATURES             Location/Qualifiers
     source          1..6762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     4..23
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        214..589
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             590..2305
                     /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(2510..2965)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2647..2668)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2859..2878
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2942..3322)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3091..3113
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3105..3122
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3106..3122
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3132..3151
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3132..3150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3159..3266
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3175..3191
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3176..3192
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3226..3242)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3226..3242)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3279..3299)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3279..3297)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3318..3334)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3318..3334)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3331..3353)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3342..3358
                     /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(3366..3372,3373..3390,3391..3396))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3411..3432
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3549..3566)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3720..4308)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3800..3819)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4479..5270,5271..5339))
                     /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     5102..5121
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5340..5444)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(5530..6333)
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        complement(6334..6549)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     6588..6606
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6644..6666)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
       61 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
      121 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
      181 cccttataaa tcaaaagaat agatctgaat taattcttga ataatacata acttttctta
      241 aaagaatcaa agacagataa aatttaagag atattaaata ttagtgagaa gccgagaatt
      301 ttgtaacacc aacataacac tgacatcttt aacaactttt aattatgata catttcttac
      361 gtcatgattg attattacag ctatgctgac aaatgactct tgttgcatgg ctacgaaccg
      421 ggtaatacta agtgattgac tcttgctgac cttttattaa gaactaaatg gacaatatta
      481 tggagcattt catgtataaa ttggtgcgta aaatcgttgg atctctcttc taagtacatc
      541 ctactataac aatcaagaaa aacaagaaaa tcggacaaaa caatcaagta tggattctag
      601 aacagttggt atattaggag ggggacaatt gggacgtatg attgttgagg cagcaaacag
      661 gctcaacatt aagacggtaa tactagatgc tgaaaattct cctgccaaac aaataagcaa
      721 ctccaatgac cacgttaatg gctccttttc caatcctctt gatatcgaaa aactagctga
      781 aaaatgtgat gtgctaacga ttgagattga gcatgttgat gttcctacac taaagaatct
      841 tcaagtaaaa catcccaaat taaaaattta cccttctcca gaaacaatca gattgataca
      901 agacaaatat attcaaaaag agcatttaat caaaaatggt atagcagtta cccaaagtgt
      961 tcctgtggaa caagccagtg agacgtccct attgaatgtt ggaagagatt tgggttttcc
     1021 attcgtcttg aagtcgagga ctttggcata cgatggaaga ggtaacttcg ttgtaaagaa
     1081 taaggaaatg attccggaag ctttggaagt actgaaggat cgtcctttgt acgccgaaaa
     1141 atgggcacca tttactaaag aattagcagt catgattgtg aggtctgtta acggtttagt
     1201 gttttcttac ccaattgtag agactatcca caaggacaat atttgtgact tatgttatgc
     1261 gcctgctaga gttccggact ccgttcaact taaggcgaag ttgttggcag aaaatgcaat
     1321 caaatctttt cccggttgtg gtatatttgg tgtggaaatg ttctatttag aaacagggga
     1381 attgcttatt aacgaaattg ccccaaggcc tcacaactct ggacattata ccattgatgc
     1441 ttgcgtcact tctcaatttg aagctcattt gagatcaata ttggatttgc caatgccaaa
     1501 gaatttcaca tctttctcca ccattacaac gaacgccatt atgctaaatg ttcttggaga
     1561 caaacataca aaagataaag agctagaaac ttgcgaaaga gcattggcga ctccaggttc
     1621 ctcagtgtac ttatatggaa aagagtctag acctaacaga aaagtaggtc acataaatat
     1681 tattgcctcc agtatggcgg aatgtgaaca aaggctgaac tacattacag gtagaactga
     1741 tattccaatc aaaatctctg tcgctcaaaa gttggacttg gaagcaatgg tcaaaccatt
     1801 ggttggaatc atcatgggat cagactctga cttgccggta atgtctgccg catgtgcggt
     1861 tttaaaagat tttggcgttc catttgaagt gacaatagtc tctgctcata gaactccaca
     1921 taggatgtca gcatatgcta tttccgcaag caagcgtgga attaaaacaa ttatcgctgg
     1981 agctggtggg gctgctcact tgccaggtat ggtggctgca atgacaccac ttcctgtcat
     2041 cggtgtgccc gtaaaaggtt cttgtctaga tggagtagat tctttacatt caattgtgca
     2101 aatgcctaga ggtgttccag tagctaccgt cgctattaat aatagtacga acgctgcgct
     2161 gttggctgtc agactgcttg gcgcttatga ttcaagttat acaacgaaaa tggaacagtt
     2221 tttattaaag caagaagaag aagttcttgt caaagcacaa aagttagaaa ctgtcggtta
     2281 cgaagcttat ctagaaaaca agtaatatat aagtttattg atatacttgt acagcaaata
     2341 attataaaat gatataccta ttttttaggc tttgttatga ttacatcaaa tgtggacttc
     2401 atacatagaa atcaacgctt acaggtgtcc ttttttaaga atttcataca taagatctat
     2461 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     2521 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     2581 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     2641 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     2701 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     2761 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     2821 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     2881 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     2941 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     3001 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3061 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3121 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg
     3181 tcgacggtat cgataagctt gatatcgaat tcctgcagcc cgggggatcc actagttcta
     3241 gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg
     3301 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3481 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4141 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4261 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4321 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4381 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4441 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4501 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4561 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4621 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4681 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4741 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4801 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4861 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4921 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4981 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5041 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5101 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5161 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5221 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5281 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5341 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5401 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5461 gtgccacctg acgtctgaag ctctaatttg tgagtttagt atacatgcat ttacttataa
     5521 tacagttttt tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgcttttct
     5581 gtaacgttca ccctctacct tagcatccct tccctttgca aatagtcctc ttccaacaat
     5641 aataatgtca gatcctgtag agaccacatc atccacggtt ctatactgtt gacccaatgc
     5701 gtctcccttg tcatctaaac ccacaccggg tgtcataatc aaccaatcgt aaccttcatc
     5761 tcttccaccc atgtctcttt gagcaataaa gccgataaca aaatctttgt cgctcttcgc
     5821 aatgtcaaca gtacccttag tatattctcc agtagatagg gagcccttgc atgacaattc
     5881 tgctaacatc aaaaggcctc taggttcctt tgttacttct tctgccgcct gcttcaaacc
     5941 gctaacaata cctgggccca ccacaccgtg tgcattcgta atgtctgccc attctgctat
     6001 tctgtataca cccgcagagt actgcaattt gactgtatta ccaatgtcag caaattttct
     6061 gtcttcgaag agtaaaaaat tgtacttggc ggataatgcc tttagcggct taactgtgcc
     6121 ctccatggaa aaatcagtca agatatccac atgtgttttt agtaaacaaa ttttgggacc
     6181 taatgcttca actaactcca gtaattcctt ggtggtacga acatccaatg aagcacacaa
     6241 gtttgtttgc ttttcgtgca tgatattaaa tagcttggca gcaacaggac taggatgagt
     6301 agcagcacgt tccttatatg tagctttcga catgatttat cttcgtttcc tgcaggtttt
     6361 tgttctgtgc agttgggtta agaatactgg gcaatttcat gtttcttcaa cactacatat
     6421 gcgtatatat accaatctaa gtctgtgctc cttccttcgt tcttccttct gttcggagat
     6481 taccgaatca aaaaaatttc aaggaaaccg aaatcaaaaa aaagaataaa aaaaaaatga
     6541 tgaattgaaa agctgtggta tggtgcatca tgacattaac ctataaaaat aggcgtatca
     6601 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     6661 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     6721 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ct
//
