LOCUS       sequence-238962-        7574 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-LEU2 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 7574)
  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 7574)
  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-LEU2
FEATURES             Location/Qualifiers
     source          1..7574
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..56
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(94..116)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     216..235
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        426..801
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             802..2517
                     /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(2722..3177)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2859..2880)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3071..3090
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(3154..3534)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3303..3325
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3317..3334
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3318..3334
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3344..3363
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3344..3362
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3371..3478
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3387..3403
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3388..3404
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3438..3454)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3438..3454)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3491..3511)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3491..3509)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3530..3546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3530..3546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3543..3565)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3554..3570
                     /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(3578..3584,3585..3602,3603..3608))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3623..3644
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3761..3778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3932..4520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4012..4031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4691..5482,5483..5551))
                     /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     5314..5333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5552..5656)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5824..6231
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6232..7326
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
ORIGIN
        1 ggcaccacac aaaaagatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
       61 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
      121 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
      181 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
      241 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
      301 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
      361 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagatctga
      421 attaattctt gaataataca taacttttct taaaagaatc aaagacagat aaaatttaag
      481 agatattaaa tattagtgag aagccgagaa ttttgtaaca ccaacataac actgacatct
      541 ttaacaactt ttaattatga tacatttctt acgtcatgat tgattattac agctatgctg
      601 acaaatgact cttgttgcat ggctacgaac cgggtaatac taagtgattg actcttgctg
      661 accttttatt aagaactaaa tggacaatat tatggagcat ttcatgtata aattggtgcg
      721 taaaatcgtt ggatctctct tctaagtaca tcctactata acaatcaaga aaaacaagaa
      781 aatcggacaa aacaatcaag tatggattct agaacagttg gtatattagg agggggacaa
      841 ttgggacgta tgattgttga ggcagcaaac aggctcaaca ttaagacggt aatactagat
      901 gctgaaaatt ctcctgccaa acaaataagc aactccaatg accacgttaa tggctccttt
      961 tccaatcctc ttgatatcga aaaactagct gaaaaatgtg atgtgctaac gattgagatt
     1021 gagcatgttg atgttcctac actaaagaat cttcaagtaa aacatcccaa attaaaaatt
     1081 tacccttctc cagaaacaat cagattgata caagacaaat atattcaaaa agagcattta
     1141 atcaaaaatg gtatagcagt tacccaaagt gttcctgtgg aacaagccag tgagacgtcc
     1201 ctattgaatg ttggaagaga tttgggtttt ccattcgtct tgaagtcgag gactttggca
     1261 tacgatggaa gaggtaactt cgttgtaaag aataaggaaa tgattccgga agctttggaa
     1321 gtactgaagg atcgtccttt gtacgccgaa aaatgggcac catttactaa agaattagca
     1381 gtcatgattg tgaggtctgt taacggttta gtgttttctt acccaattgt agagactatc
     1441 cacaaggaca atatttgtga cttatgttat gcgcctgcta gagttccgga ctccgttcaa
     1501 cttaaggcga agttgttggc agaaaatgca atcaaatctt ttcccggttg tggtatattt
     1561 ggtgtggaaa tgttctattt agaaacaggg gaattgctta ttaacgaaat tgccccaagg
     1621 cctcacaact ctggacatta taccattgat gcttgcgtca cttctcaatt tgaagctcat
     1681 ttgagatcaa tattggattt gccaatgcca aagaatttca catctttctc caccattaca
     1741 acgaacgcca ttatgctaaa tgttcttgga gacaaacata caaaagataa agagctagaa
     1801 acttgcgaaa gagcattggc gactccaggt tcctcagtgt acttatatgg aaaagagtct
     1861 agacctaaca gaaaagtagg tcacataaat attattgcct ccagtatggc ggaatgtgaa
     1921 caaaggctga actacattac aggtagaact gatattccaa tcaaaatctc tgtcgctcaa
     1981 aagttggact tggaagcaat ggtcaaacca ttggttggaa tcatcatggg atcagactct
     2041 gacttgccgg taatgtctgc cgcatgtgcg gttttaaaag attttggcgt tccatttgaa
     2101 gtgacaatag tctctgctca tagaactcca cataggatgt cagcatatgc tatttccgca
     2161 agcaagcgtg gaattaaaac aattatcgct ggagctggtg gggctgctca cttgccaggt
     2221 atggtggctg caatgacacc acttcctgtc atcggtgtgc ccgtaaaagg ttcttgtcta
     2281 gatggagtag attctttaca ttcaattgtg caaatgccta gaggtgttcc agtagctacc
     2341 gtcgctatta ataatagtac gaacgctgcg ctgttggctg tcagactgct tggcgcttat
     2401 gattcaagtt atacaacgaa aatggaacag tttttattaa agcaagaaga agaagttctt
     2461 gtcaaagcac aaaagttaga aactgtcggt tacgaagctt atctagaaaa caagtaatat
     2521 ataagtttat tgatatactt gtacagcaaa taattataaa atgatatacc tattttttag
     2581 gctttgttat gattacatca aatgtggact tcatacatag aaatcaacgc ttacaggtgt
     2641 ccttttttaa gaatttcata cataagatct atgcggtgtg aaataccgca cagatgcgta
     2701 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2761 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2821 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2881 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2941 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3001 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3061 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3121 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     3181 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     3241 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     3301 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     3361 gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga
     3421 attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     3481 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     3541 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3601 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3661 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3721 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3781 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3841 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3901 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3961 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4021 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4081 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4141 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4201 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4261 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4321 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4381 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4441 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4501 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4561 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4621 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4681 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4741 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4801 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4861 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4921 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4981 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5041 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5101 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5161 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5221 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5281 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5341 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5401 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5461 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5521 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5581 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5641 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctcc tgtacttcct
     5701 tgttcatgtg tgttcaaaaa cgttatattt ataggataat tatactctat ttctcaacaa
     5761 gtaattggtt gtttggccga gcggtctaag gcgcctgatt caagaaatat cttgaccgca
     5821 gttaactgtg ggaatactca ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca
     5881 ttattttttt cctcaacata acgagaacac acaggggcgc tatcgcacag aatcaaattc
     5941 gatgactgga aattttttgt taatttcaga ggtcgcctga cgcatatacc tttttcaact
     6001 gaaaaattgg gagaaaaagg aaaggtgaga gcgccggaac cggcttttca tatagaatag
     6061 agaagcgttc atgactaaat gcttgcatca caatacttga agttgacaat attatttaag
     6121 gacctattgt tttttccaat aggtggttag caatcgtctt actttctaac ttttcttacc
     6181 ttttacattt cagcaatata tatatatata tttcaaggat ataccattct aatgtctgcc
     6241 cctaagaaga tcgtcgtttt gccaggtgac cacgttggtc aagaaatcac agccgaagcc
     6301 attaaggttc ttaaagctat ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat
     6361 catttaattg gtggtgctgc tatcgatgct acaggtgttc cacttccaga tgaggcgctg
     6421 gaagcctcca agaaggctga tgccgttttg ttaggtgctg tgggtggtcc taaatggggt
     6481 accggtagtg ttagacctga acaaggttta ctaaaaatcc gtaaagaact tcaattgtac
     6541 gccaacttaa gaccatgtaa ctttgcatcc gactctcttt tagacttatc tccaatcaag
     6601 ccacaatttg ctaaaggtac tgacttcgtt gttgtcagag aattagtggg aggtatttac
     6661 tttggtaaga gaaaggaaga cgatggtgat ggtgtcgctt gggatagtga acaatacacc
     6721 gttccagaag tgcaaagaat cacaagaatg gccgctttca tggccctaca acatgagcca
     6781 ccattgccta tttggtcctt ggataaagct aatgttttgg cctcttcaag attatggaga
     6841 aaaactgtgg aggaaaccat caagaacgaa ttccctacat tgaaggttca acatcaattg
     6901 attgattctg ccgccatgat cctagttaag aacccaaccc acctaaatgg tattataatc
     6961 accagcaaca tgtttggtga tatcatctcc gatgaagcct ccgttatccc aggttccttg
     7021 ggtttgttgc catctgcgtc cttggcctct ttgccagaca agaacaccgc atttggtttg
     7081 tacgaaccat gccacggttc tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc
     7141 actatcttgt ctgctgcaat gatgttgaaa ttgtcattga acttgcctga agaaggtaag
     7201 gccattgaag atgcagttaa aaaggttttg gatgcaggta tcagaactgg tgatttaggt
     7261 ggttccaaca gtaccaccga agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt
     7321 gcttaaaaag attctctttt tttatgatat ttgtacataa actttataaa tgaaattcat
     7381 aatagaaacg acacgaaatt acaaaatgga atatgttcat agggtagacg aaactatata
     7441 cgcaatctac atacatttat caagaaggag aaaaaggagg atgtaaagga atacaggtaa
     7501 gcaaattgat actaatggct caacgtgata aggaaaaaga attgcacttt aacattaata
     7561 ttgacaagga ggag
//
