LOCUS       sequence-239186-        6863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-NatMx 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 6863)
  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 6863)
  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-NatMx
FEATURES             Location/Qualifiers
     source          1..6863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        134..509
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             510..2225
                     /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(2430..2885)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2567..2588)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2779..2798
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2862..3242)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3011..3033
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3025..3042
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3026..3042
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3052..3071
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3052..3070
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3079..3186
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3095..3111
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3096..3112
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3146..3162)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3146..3162)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3199..3219)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3199..3217)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3238..3254)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3238..3254)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3251..3273)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3262..3278
                     /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(3286..3292,3293..3310,3311..3316))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3331..3352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3469..3486)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3640..4228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3720..3739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4399..5190,5191..5259))
                     /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     5022..5041
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5260..5364)
                     /gene="bla"
                     /label=AmpR promoter
     gene            5437..6556
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        5437..5780
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5781..6353
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      6359..6556
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     6609..6627
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6665..6687)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6787..6806
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
       61 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      121 agatctgaat taattcttga ataatacata acttttctta aaagaatcaa agacagataa
      181 aatttaagag atattaaata ttagtgagaa gccgagaatt ttgtaacacc aacataacac
      241 tgacatcttt aacaactttt aattatgata catttcttac gtcatgattg attattacag
      301 ctatgctgac aaatgactct tgttgcatgg ctacgaaccg ggtaatacta agtgattgac
      361 tcttgctgac cttttattaa gaactaaatg gacaatatta tggagcattt catgtataaa
      421 ttggtgcgta aaatcgttgg atctctcttc taagtacatc ctactataac aatcaagaaa
      481 aacaagaaaa tcggacaaaa caatcaagta tggattctag aacagttggt atattaggag
      541 ggggacaatt gggacgtatg attgttgagg cagcaaacag gctcaacatt aagacggtaa
      601 tactagatgc tgaaaattct cctgccaaac aaataagcaa ctccaatgac cacgttaatg
      661 gctccttttc caatcctctt gatatcgaaa aactagctga aaaatgtgat gtgctaacga
      721 ttgagattga gcatgttgat gttcctacac taaagaatct tcaagtaaaa catcccaaat
      781 taaaaattta cccttctcca gaaacaatca gattgataca agacaaatat attcaaaaag
      841 agcatttaat caaaaatggt atagcagtta cccaaagtgt tcctgtggaa caagccagtg
      901 agacgtccct attgaatgtt ggaagagatt tgggttttcc attcgtcttg aagtcgagga
      961 ctttggcata cgatggaaga ggtaacttcg ttgtaaagaa taaggaaatg attccggaag
     1021 ctttggaagt actgaaggat cgtcctttgt acgccgaaaa atgggcacca tttactaaag
     1081 aattagcagt catgattgtg aggtctgtta acggtttagt gttttcttac ccaattgtag
     1141 agactatcca caaggacaat atttgtgact tatgttatgc gcctgctaga gttccggact
     1201 ccgttcaact taaggcgaag ttgttggcag aaaatgcaat caaatctttt cccggttgtg
     1261 gtatatttgg tgtggaaatg ttctatttag aaacagggga attgcttatt aacgaaattg
     1321 ccccaaggcc tcacaactct ggacattata ccattgatgc ttgcgtcact tctcaatttg
     1381 aagctcattt gagatcaata ttggatttgc caatgccaaa gaatttcaca tctttctcca
     1441 ccattacaac gaacgccatt atgctaaatg ttcttggaga caaacataca aaagataaag
     1501 agctagaaac ttgcgaaaga gcattggcga ctccaggttc ctcagtgtac ttatatggaa
     1561 aagagtctag acctaacaga aaagtaggtc acataaatat tattgcctcc agtatggcgg
     1621 aatgtgaaca aaggctgaac tacattacag gtagaactga tattccaatc aaaatctctg
     1681 tcgctcaaaa gttggacttg gaagcaatgg tcaaaccatt ggttggaatc atcatgggat
     1741 cagactctga cttgccggta atgtctgccg catgtgcggt tttaaaagat tttggcgttc
     1801 catttgaagt gacaatagtc tctgctcata gaactccaca taggatgtca gcatatgcta
     1861 tttccgcaag caagcgtgga attaaaacaa ttatcgctgg agctggtggg gctgctcact
     1921 tgccaggtat ggtggctgca atgacaccac ttcctgtcat cggtgtgccc gtaaaaggtt
     1981 cttgtctaga tggagtagat tctttacatt caattgtgca aatgcctaga ggtgttccag
     2041 tagctaccgt cgctattaat aatagtacga acgctgcgct gttggctgtc agactgcttg
     2101 gcgcttatga ttcaagttat acaacgaaaa tggaacagtt tttattaaag caagaagaag
     2161 aagttcttgt caaagcacaa aagttagaaa ctgtcggtta cgaagcttat ctagaaaaca
     2221 agtaatatat aagtttattg atatacttgt acagcaaata attataaaat gatataccta
     2281 ttttttaggc tttgttatga ttacatcaaa tgtggacttc atacatagaa atcaacgctt
     2341 acaggtgtcc ttttttaaga atttcataca taagatctat gcggtgtgaa ataccgcaca
     2401 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     2461 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2521 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2581 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2641 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2701 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2761 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2821 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2881 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     2941 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     3001 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac
     3061 tcactatagg gcgaattggg taccgggccc cccctcgagg tcgacggtat cgataagctt
     3121 gatatcgaat tcctgcagcc cgggggatcc actagttcta gagcggccgc caccgcggtg
     3181 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     3241 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     3301 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     3361 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3421 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3481 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3541 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3601 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3661 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3721 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3781 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3841 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3901 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3961 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4021 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4081 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4141 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4201 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4261 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4321 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4381 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4441 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4501 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4561 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4621 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4681 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4741 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4801 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4861 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4921 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4981 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5041 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5101 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5161 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5221 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5281 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5341 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcttgtt
     5401 tagcttgcct cgtccccgcc gggtcacccg gccagcgaca tggaggccca gaataccctc
     5461 cttgacagtc ttgacgtgcg cagctcaggg gcatgatgtg actgtcgccc gtacatttag
     5521 cccatacatc cccatgtata atcatttgca tccatacatt ttgatggccg cacggcgcga
     5581 agcaaaaatt acggctcctc gctgcagacc tgcgagcagg gaaacgctcc cctcacagac
     5641 gcgttgaatt gtccccacgc cgcgcccctg tagagaaata taaaaggtta ggatttgcca
     5701 ctgaggttct tctttcatat acttcctttt aaaatcttgc taggatacag ttctcacatc
     5761 acatccgaac ataaacaacc atgggtacca ctcttgacga cacggcttac cggtaccgca
     5821 ccagtgtccc gggggacgcc gaggccatcg aggcactgga tgggtccttc accaccgaca
     5881 ccgtcttccg cgtcaccgcc accggggacg gcttcaccct gcgggaggtg ccggtggacc
     5941 cgcccctgac caaggtgttc cccgacgacg aatcggacga cgaatcggac gacggggagg
     6001 acggcgaccc ggactcccgg acgttcgtcg cgtacgggga cgacggcgac ctggcgggct
     6061 tcgtggtcgt ctcgtactcc ggctggaacc gccggctgac cgtcgaggac atcgaggtcg
     6121 ccccggagca ccgggggcac ggggtcgggc gcgcgttgat ggggctcgcg acggagttcg
     6181 cccgcgagcg gggcgccggg cacctctggc tggaggtcac caacgtcaac gcaccggcga
     6241 tccacgcgta ccggcggatg gggttcaccc tctgcggcct ggacaccgcc ctgtacgacg
     6301 gcaccgcctc ggacggcgag caggcgctct acatgagcat gccctgcccc taatcagtac
     6361 tgacaataaa aagattcttg ttttcaagaa cttgtcattt gtatagtttt tttatattgt
     6421 agttgttcta ttttaatcaa atgttagcgt gatttatatt ttttttcgcc tcgacatcat
     6481 ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca tgcgtcaatc gtatgtgaat
     6541 gctggtcgct atactgctgt cgattcgata ctaacgccgc catccagatc atgacattaa
     6601 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     6661 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     6721 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     6781 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     6841 acagatgcgt aaggagaaaa tac
//
