LOCUS       sequence-238934-        7260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-MET17 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 7260)
  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 7260)
  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-MET17
FEATURES             Location/Qualifiers
     source          1..7260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             258..1592
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
     promoter        1643..1747
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1748..1816,1817..2608)
                     /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     complement(1966..1985)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2779..3367
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3268..3287
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3521..3538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3655..3676
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3691..3696,3697..3714,3715..3721)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3729..3745
                     /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)."
     primer_bind     3734..3756
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3753..3769
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3753..3769
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3765..4145
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3788..3808
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3790..3808
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    3821..3928
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3845..3861
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3845..3861
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3895..3911)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3896..3912)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3936..3955)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3937..3955)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3965..3982)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3965..3981)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3974..3996)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4122..4577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4209..4228)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4419..4440
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4782..6497)
                     /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"
     promoter        complement(6498..6873)
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     primer_bind     complement(7064..7083)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7183..7205
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(join(7243..7260,1))
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttttataggt taatgtcatg atagccatcc tcatgaaaac tgtgtaacat aataaccgaa
       61 gtgtcgaaaa ggtggcacct tgtccaattg aacacgctcg atgaaaaaaa taagatatat
      121 ataaggttaa gtaaagcgtc tgttagaaag gaagtttttc ctttttcttg ctctcttgtc
      181 ttttcatcta ctatttcctt cgtgtaatac agggtcgtca gatacataga tacaattcta
      241 ttacccccat ccatacaatg ccatctcatt tcgatactgt tcaactacac gccggccaag
      301 agaaccctgg tgacaatgct cacagatcca gagctgtacc aatttacgcc accacttctt
      361 atgttttcga aaactctaag catggttcgc aattgtttgg tctagaagtt ccaggttacg
      421 tctattcccg tttccaaaac ccaaccagta atgttttgga agaaagaatt gctgctttag
      481 aaggtggtgc tgctgctttg gctgtttcct ccggtcaagc cgctcaaacc cttgccatcc
      541 aaggtttggc acacactggt gacaacatcg tttccacttc ttacttatac ggtggtactt
      601 ataaccagtt caaaatctcg ttcaaaagat ttggtatcga ggctagattt gttgaaggtg
      661 acaatccaga agaattcgaa aaggtctttg atgaaagaac caaggctgtt tatttggaaa
      721 ccattggtaa tccaaagtac aatgttccgg attttgaaaa aattgttgca attgctcaca
      781 aacacggtat tccagttgtc gttgacaaca catttggtgc cggtggttac ttctgtcagc
      841 caattaaata cggtgctgat attgtaacac attctgctac caaatggatt ggtggtcatg
      901 gtactactat cggtggtatt attgttgact ctggtaagtt cccatggaag gactacccag
      961 aaaagttccc tcaattctct caacctgccg aaggatatca cggtactatc tacaatgaag
     1021 cctacggtaa cttggcatac atcgttcatg ttagaactga actattaaga gatttgggtc
     1081 cattgatgaa cccatttgcc tctttcttgc tactacaagg tgttgaaaca ttatctttga
     1141 gagctgaaag acacggtgaa aatgcattga agttagccaa atggttagaa caatccccat
     1201 acgtatcttg ggtttcatac cctggtttag catctcattc tcatcatgaa aatgctaaga
     1261 agtatctatc taacggtttc ggtggtgtct tatctttcgg tgtaaaagac ttaccaaatg
     1321 ccgacaagga aactgaccca ttcaaacttt ctggtgctca agttgttgac aatttaaagc
     1381 ttgcctctaa cttggccaat gttggtgatg ccaagacctt agtcattgct ccatacttca
     1441 ctacccacaa acaattaaat gacaaagaaa agttggcatc tggtgttacc aaggacttaa
     1501 ttcgtgtctc tgttggtatc gaatttattg atgacattat tgcagacttc cagcaatctt
     1561 ttgaaactgt tttcgctggc caaaaaccat gagtgtgcgt aatgagttgt agacgtcagg
     1621 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1681 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1741 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1801 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1861 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1921 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1981 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2041 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2101 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2161 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2221 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2281 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2341 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2401 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2461 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2521 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2581 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2641 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2701 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2761 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2821 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2881 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2941 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3001 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3061 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3121 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3181 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3241 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3301 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3361 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3421 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3481 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3541 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3601 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     3661 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     3721 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     3781 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg
     3841 ccgctctaga actagtggat cccccgggct gcaggaattc gatatcaagc ttatcgatac
     3901 cgtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg tattacgcgc
     3961 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     4021 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     4081 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     4141 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4201 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4261 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4321 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4381 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4441 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4501 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4561 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4621 caccgcatag atcttatgta tgaaattctt aaaaaaggac acctgtaagc gttgatttct
     4681 atgtatgaag tccacatttg atgtaatcat aacaaagcct aaaaaatagg tatatcattt
     4741 tataattatt tgctgtacaa gtatatcaat aaacttatat attacttgtt ttctagataa
     4801 gcttcgtaac cgacagtttc taacttttgt gctttgacaa gaacttcttc ttcttgcttt
     4861 aataaaaact gttccatttt cgttgtataa cttgaatcat aagcgccaag cagtctgaca
     4921 gccaacagcg cagcgttcgt actattatta atagcgacgg tagctactgg aacacctcta
     4981 ggcatttgca caattgaatg taaagaatct actccatcta gacaagaacc ttttacgggc
     5041 acaccgatga caggaagtgg tgtcattgca gccaccatac ctggcaagtg agcagcccca
     5101 ccagctccag cgataattgt tttaattcca cgcttgcttg cggaaatagc atatgctgac
     5161 atcctatgtg gagttctatg agcagagact attgtcactt caaatggaac gccaaaatct
     5221 tttaaaaccg cacatgcggc agacattacc ggcaagtcag agtctgatcc catgatgatt
     5281 ccaaccaatg gtttgaccat tgcttccaag tccaactttt gagcgacaga gattttgatt
     5341 ggaatatcag ttctacctgt aatgtagttc agcctttgtt cacattccgc catactggag
     5401 gcaataatat ttatgtgacc tacttttctg ttaggtctag actcttttcc atataagtac
     5461 actgaggaac ctggagtcgc caatgctctt tcgcaagttt ctagctcttt atcttttgta
     5521 tgtttgtctc caagaacatt tagcataatg gcgttcgttg taatggtgga gaaagatgtg
     5581 aaattctttg gcattggcaa atccaatatt gatctcaaat gagcttcaaa ttgagaagtg
     5641 acgcaagcat caatggtata atgtccagag ttgtgaggcc ttggggcaat ttcgttaata
     5701 agcaattccc ctgtttctaa atagaacatt tccacaccaa atataccaca accgggaaaa
     5761 gatttgattg cattttctgc caacaacttc gccttaagtt gaacggagtc cggaactcta
     5821 gcaggcgcat aacataagtc acaaatattg tccttgtgga tagtctctac aattgggtaa
     5881 gaaaacacta aaccgttaac agacctcaca atcatgactg ctaattcttt agtaaatggt
     5941 gcccattttt cggcgtacaa aggacgatcc ttcagtactt ccaaagcttc cggaatcatt
     6001 tccttattct ttacaacgaa gttacctctt ccatcgtatg ccaaagtcct cgacttcaag
     6061 acgaatggaa aacccaaatc tcttccaaca ttcaataggg acgtctcact ggcttgttcc
     6121 acaggaacac tttgggtaac tgctatacca tttttgatta aatgctcttt ttgaatatat
     6181 ttgtcttgta tcaatctgat tgtttctgga gaagggtaaa tttttaattt gggatgtttt
     6241 acttgaagat tctttagtgt aggaacatca acatgctcaa tctcaatcgt tagcacatca
     6301 catttttcag ctagtttttc gatatcaaga ggattggaaa aggagccatt aacgtggtca
     6361 ttggagttgc ttatttgttt ggcaggagaa ttttcagcat ctagtattac cgtcttaatg
     6421 ttgagcctgt ttgctgcctc aacaatcata cgtcccaatt gtccccctcc taatatacca
     6481 actgttctag aatccatact tgattgtttt gtccgatttt cttgtttttc ttgattgtta
     6541 tagtaggatg tacttagaag agagatccaa cgattttacg caccaattta tacatgaaat
     6601 gctccataat attgtccatt tagttcttaa taaaaggtca gcaagagtca atcacttagt
     6661 attacccggt tcgtagccat gcaacaagag tcatttgtca gcatagctgt aataatcaat
     6721 catgacgtaa gaaatgtatc ataattaaaa gttgttaaag atgtcagtgt tatgttggtg
     6781 ttacaaaatt ctcggcttct cactaatatt taatatctct taaattttat ctgtctttga
     6841 ttcttttaag aaaagttatg tattattcaa gaattaattc agatctattc ttttgattta
     6901 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     6961 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     7021 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     7081 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     7141 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     7201 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
//
