LOCUS       sequence-238940-        5372 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-URA3 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 5372)
  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 5372)
  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-URA3
FEATURES             Location/Qualifiers
     source          1..5372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..121
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        145..360
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             361..1164
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1295..1750)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1432..1453)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1644..1663
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1727..2107)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1876..1898
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1890..1907
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1891..1907
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1917..1936
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1917..1935
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1944..2051
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1960..1976
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1961..1977
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2011..2027)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2011..2027)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2064..2084)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2064..2082)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2103..2119)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2103..2119)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2116..2138)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2127..2143
                     /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(2151..2157,2158..2175,2176..2181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2196..2217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2334..2351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2505..3093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2585..2604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3264..4055,4056..4124))
                     /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     3887..3906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4125..4229)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4270..4551
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             4552..5226
                     /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"
     primer_bind     5296..5314
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(5352..5372,1..2))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
       61 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
      121 actgagagtg caccatacca ccttttcaat tcatcatttt ttttttattc ttttttttga
      181 tttcggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
      241 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
      301 tgcccagtat tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc
      361 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
      421 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
      481 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
      541 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
      601 tccgccaagt acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca
      661 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
      721 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
      781 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
      841 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
      901 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
      961 acacccggtg tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg
     1021 gatgatgtgg tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     1081 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     1141 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     1201 tcacaaatta gagcttcaat ttaattatat cagttattac cctatgcggt gtgaaatacc
     1261 gcacagatgc gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta
     1321 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     1381 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     1441 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     1501 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     1561 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     1621 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     1681 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     1741 cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     1801 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1861 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     1921 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac ggtatcgata
     1981 agcttgatat cgaattcctg cagcccgggg gatccactag ttctagagcg gccgccaccg
     2041 cggtggagct ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     2101 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2161 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     2221 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     2281 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2341 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2401 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2461 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2521 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2581 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2641 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2701 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2761 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2821 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2881 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2941 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3001 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3061 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3121 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3181 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3241 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3301 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3361 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3421 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3481 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3541 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3601 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3661 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3721 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3781 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3841 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3901 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3961 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4021 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4081 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4141 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4201 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4261 tgcaccataa acgacattac tatatatata atataggaag catttaatag aacagcatcg
     4321 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat
     4381 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga
     4441 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
     4501 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
     4561 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
     4621 gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat
     4681 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
     4741 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
     4801 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
     4861 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
     4921 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
     4981 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg
     5041 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca
     5101 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt
     5161 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag
     5221 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg cctaatcatg
     5281 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     5341 gacggtgaaa acctctgaca catgcagctc cc
//
