LOCUS       sequence-239163-        7100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-KanMx 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 7100)
  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 7100)
  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-KanMx
FEATURES             Location/Qualifiers
     source          1..7100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     67..86
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        277..652
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             653..2368
                     /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(2573..3028)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2710..2731)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2922..2941
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(3005..3385)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3154..3176
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3168..3185
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3169..3185
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3195..3214
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3195..3213
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3222..3329
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3238..3254
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3239..3255
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3289..3305)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3289..3305)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3342..3362)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3342..3360)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3381..3397)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3381..3397)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3394..3416)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3405..3421
                     /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(3429..3435,3436..3453,3454..3459))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3474..3495
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3612..3629)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3783..4371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3863..3882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4542..5333,5334..5402))
                     /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     5165..5184
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5403..5507)
                     /gene="bla"
                     /label=AmpR promoter
     gene            5580..6936
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        5580..5923
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5924..6733
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5991..6010)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      6739..6936
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     6989..7007
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7045..7067)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
       61 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
      121 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
      181 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
      241 aatcccttat aaatcaaaag aatagatctg aattaattct tgaataatac ataacttttc
      301 ttaaaagaat caaagacaga taaaatttaa gagatattaa atattagtga gaagccgaga
      361 attttgtaac accaacataa cactgacatc tttaacaact tttaattatg atacatttct
      421 tacgtcatga ttgattatta cagctatgct gacaaatgac tcttgttgca tggctacgaa
      481 ccgggtaata ctaagtgatt gactcttgct gaccttttat taagaactaa atggacaata
      541 ttatggagca tttcatgtat aaattggtgc gtaaaatcgt tggatctctc ttctaagtac
      601 atcctactat aacaatcaag aaaaacaaga aaatcggaca aaacaatcaa gtatggattc
      661 tagaacagtt ggtatattag gagggggaca attgggacgt atgattgttg aggcagcaaa
      721 caggctcaac attaagacgg taatactaga tgctgaaaat tctcctgcca aacaaataag
      781 caactccaat gaccacgtta atggctcctt ttccaatcct cttgatatcg aaaaactagc
      841 tgaaaaatgt gatgtgctaa cgattgagat tgagcatgtt gatgttccta cactaaagaa
      901 tcttcaagta aaacatccca aattaaaaat ttacccttct ccagaaacaa tcagattgat
      961 acaagacaaa tatattcaaa aagagcattt aatcaaaaat ggtatagcag ttacccaaag
     1021 tgttcctgtg gaacaagcca gtgagacgtc cctattgaat gttggaagag atttgggttt
     1081 tccattcgtc ttgaagtcga ggactttggc atacgatgga agaggtaact tcgttgtaaa
     1141 gaataaggaa atgattccgg aagctttgga agtactgaag gatcgtcctt tgtacgccga
     1201 aaaatgggca ccatttacta aagaattagc agtcatgatt gtgaggtctg ttaacggttt
     1261 agtgttttct tacccaattg tagagactat ccacaaggac aatatttgtg acttatgtta
     1321 tgcgcctgct agagttccgg actccgttca acttaaggcg aagttgttgg cagaaaatgc
     1381 aatcaaatct tttcccggtt gtggtatatt tggtgtggaa atgttctatt tagaaacagg
     1441 ggaattgctt attaacgaaa ttgccccaag gcctcacaac tctggacatt ataccattga
     1501 tgcttgcgtc acttctcaat ttgaagctca tttgagatca atattggatt tgccaatgcc
     1561 aaagaatttc acatctttct ccaccattac aacgaacgcc attatgctaa atgttcttgg
     1621 agacaaacat acaaaagata aagagctaga aacttgcgaa agagcattgg cgactccagg
     1681 ttcctcagtg tacttatatg gaaaagagtc tagacctaac agaaaagtag gtcacataaa
     1741 tattattgcc tccagtatgg cggaatgtga acaaaggctg aactacatta caggtagaac
     1801 tgatattcca atcaaaatct ctgtcgctca aaagttggac ttggaagcaa tggtcaaacc
     1861 attggttgga atcatcatgg gatcagactc tgacttgccg gtaatgtctg ccgcatgtgc
     1921 ggttttaaaa gattttggcg ttccatttga agtgacaata gtctctgctc atagaactcc
     1981 acataggatg tcagcatatg ctatttccgc aagcaagcgt ggaattaaaa caattatcgc
     2041 tggagctggt ggggctgctc acttgccagg tatggtggct gcaatgacac cacttcctgt
     2101 catcggtgtg cccgtaaaag gttcttgtct agatggagta gattctttac attcaattgt
     2161 gcaaatgcct agaggtgttc cagtagctac cgtcgctatt aataatagta cgaacgctgc
     2221 gctgttggct gtcagactgc ttggcgctta tgattcaagt tatacaacga aaatggaaca
     2281 gtttttatta aagcaagaag aagaagttct tgtcaaagca caaaagttag aaactgtcgg
     2341 ttacgaagct tatctagaaa acaagtaata tataagttta ttgatatact tgtacagcaa
     2401 ataattataa aatgatatac ctatttttta ggctttgtta tgattacatc aaatgtggac
     2461 ttcatacata gaaatcaacg cttacaggtg tcctttttta agaatttcat acataagatc
     2521 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta
     2581 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2641 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     2701 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2761 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     2821 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     2881 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     2941 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3001 gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg
     3061 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     3121 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     3181 ccagtgagcg cgcgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg
     3241 aggtcgacgg tatcgataag cttgatatcg aattcctgca gcccggggga tccactagtt
     3301 ctagagcggc cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt
     3361 gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3421 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3481 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3541 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3601 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3661 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3721 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3781 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3841 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3901 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3961 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4021 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4081 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4141 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4201 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4261 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4321 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4381 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4441 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4501 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4561 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4621 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4681 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4741 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4801 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4861 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4921 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4981 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5041 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5101 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5161 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5221 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5281 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5341 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5401 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5461 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5521 aaagtgccac ctgacgtctt gtttagcttg cctcgtcccc gccgggtcac ccggccagcg
     5581 acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca ggggcatgat
     5641 gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt gcatccatac
     5701 attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag acctgcgagc
     5761 agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc ctgtagagaa
     5821 atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct tttaaaatct
     5881 tgctaggata cagttctcac atcacatccg aacataaaca accatgggta aggaaaagac
     5941 tcacgtttcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     6001 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     6061 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     6121 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     6181 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggcaaaa cagcattcca
     6241 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     6301 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     6361 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     6421 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc ttttgccatt
     6481 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     6541 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     6601 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     6661 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     6721 tgagtttttc taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt
     6781 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     6841 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     6901 tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc
     6961 catccagatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     7021 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     7081 ttgtctgtaa gcggatgccg
//
