LOCUS       sequence-239162-        5687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-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 5687)
  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 5687)
  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 TRP1-KanMx
FEATURES             Location/Qualifiers
     source          1..5687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..136
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        157..438
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             439..1113
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1210..1665)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1347..1368)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1559..1578
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1642..2022)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1791..1813
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1805..1822
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1806..1822
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1832..1851
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1832..1850
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1859..1966
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1875..1891
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1876..1892
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1926..1942)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1926..1942)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1979..1999)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1979..1997)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2018..2034)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2018..2034)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2031..2053)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2042..2058
                     /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(2066..2072,2073..2090,2091..2096))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2111..2132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2249..2266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2420..3008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2500..2519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3179..3970,3971..4039))
                     /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     3802..3821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4040..4144)
                     /gene="bla"
                     /label=AmpR promoter
     gene            4217..5573
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        4217..4560
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             4561..5370
                     /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(4628..4647)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      5376..5573
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     5626..5644
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(5682..5687,1..17))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
       61 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
      121 atcagagcag attgtactga gagtgcacca tagatcaacg acattactat atatataata
      181 taggaagcat ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt atgacgccag
      241 atggcagtag tggaagatat tctttattga aaaatagctt gtcaccttac gtacaatctt
      301 gatccggagc ttttcttttt ttgccgatta agaattaatt cggtcgaaaa aagaaaagga
      361 gagggccaag agggagggca ttggtgacta ttgagcacgt gagtatacgt gattaagcac
      421 acaaaggcag cttggagtat gtctgttatt aatttcacag gtagttctgg tccattggtg
      481 aaagtttgcg gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac
      541 ttgctgggta ttatatgtgt gcccaataga aagagaacaa ttgacccggt tattgcaagg
      601 aaaatttcaa gtcttgtaaa agcatataaa aatagttcag gcactccgaa atacttggtt
      661 ggcgtgtttc gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt
      721 gatatcgtcc aactgcatgg agatgagtcg tggcaagaat accaagagtt cctcggtttg
      781 ccagttatta aaagactcgt atttccaaaa gactgcaaca tactactcag tgcagcttca
      841 cagaaacctc attcgtttat tcccttgttt gattcagaag caggtgggac aggtgaactt
      901 ttggattgga actcgatttc tgactgggtt ggaaggcaag agagccccga aagcttacat
      961 tttatgttag ctggtggact gacgccagaa aatgttggtg atgcgcttag attaaatggc
     1021 gttattggtg ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa
     1081 atagcaaatt tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta tttatttaat
     1141 tatatcagtt attaccctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     1201 cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     1261 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     1321 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     1381 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     1441 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     1501 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     1561 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     1621 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag
     1681 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     1741 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     1801 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
     1861 taccgggccc cccctcgagg tcgacggtat cgataagctt gatatcgaat tcctgcagcc
     1921 cgggggatcc actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
     1981 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2041 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2101 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2161 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2221 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2281 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2341 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2401 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2461 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2521 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2581 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2641 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2701 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2761 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2821 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2881 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2941 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3001 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3061 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3121 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3181 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3241 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3301 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3361 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     3421 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     3481 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     3541 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3601 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3661 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3721 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3781 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3841 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3901 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3961 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4021 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4081 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4141 cgcgcacatt tccccgaaaa gtgccacctg acgtcttgtt tagcttgcct cgtccccgcc
     4201 gggtcacccg gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg
     4261 cagctcaggg gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata
     4321 atcatttgca tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc
     4381 gctgcagacc tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc
     4441 cgcgcccctg tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat
     4501 acttcctttt aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc
     4561 atgggtaagg aaaagactca cgtttcgagg ccgcgattaa attccaacat ggatgctgat
     4621 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
     4681 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     4741 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     4801 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     4861 ggcaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     4921 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     4981 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     5041 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     5101 cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     5161 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     5221 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     5281 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     5341 tttcatttga tgctcgatga gtttttctaa tcagtactga caataaaaag attcttgttt
     5401 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg
     5461 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg
     5521 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga
     5581 ttcgatacta acgccgccat ccagatcatg acattaacct ataaaaatag gcgtatcacg
     5641 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctg
//
