LOCUS       sequence-239074-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-HIS3 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 5455)
  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 5455)
  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-HIS3
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        186..373
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             374..1036
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1296..1751)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1433..1454)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1645..1664
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1728..2108)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1877..1899
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1891..1908
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1892..1908
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1918..1937
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1918..1936
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1945..2052
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1961..1977
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1962..1978
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2012..2028)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2012..2028)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2065..2085)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2065..2083)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2104..2120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2104..2120)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2117..2139)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2128..2144
                     /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(2152..2158,2159..2176,2177..2182))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2197..2218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2335..2352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2506..3094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2586..2605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3265..4056,4057..4125))
                     /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     3888..3907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4126..4230)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4271..4552
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             4553..5227
                     /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     5297..5315
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5353..5375)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccataaattc
       61 ccgttttaag agcttggtga gcgctaggag tcactgccag gtatcgtttg aacacggcat
      121 tagtcaggga agtcataaca cagtcctttc ccgcaatttt ctttttctat tactcttggc
      181 ctcctctagt acactctata tttttttatg cctcggtaat gattttcatt tttttttttc
      241 cacctagcgg atgactcttt ttttttctta gcgattggca ttatcacata atgaattata
      301 cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc aggcaagata
      361 aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa atgaaaccaa
      421 gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact cgatcttccc
      481 agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga ttaacgtcca
      541 cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt ccggctggtc
      601 gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca ctgaagactg
      661 cgggattgct ctcggtcaag cttttaaaga ggccctaggg gccgtgcgtg gagtaaaaag
      721 gtttggatca ggatttgcgc ctttggatga ggcactttcc agagcggtgg tagatctttc
      781 gaacaggccg tacgcagttg tcgaacttgg tttgcaaagg gagaaagtag gagatctctc
      841 ttgcgagatg atcccgcatt ttcttgaaag ctttgcagag gctagcagaa ttaccctcca
      901 cgttgattgt ctgcgaggca agaatgatca tcaccgtagt gagagtgcgt tcaaggctct
      961 tgcggttgcc ataagagaag ccacctcgcc caatggtacc aacgatgttc cctccaccaa
     1021 aggtgttctt atgtagtgac accgattatt taaagctgca gcatacgata tatatacatg
     1081 tgtatatatg tatacctatg aatgtcagta agtatgtata cgaacagtat gatactgaag
     1141 atgacaaggt aatgcatcat tctatacgtg tcattctgaa cgaggcgcgc tttccttttt
     1201 tctttttgct ttttcttttt ttttctcttg aactcgacgg atctatgcgg tgtgaaatac
     1261 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     1321 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     1381 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     1441 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     1501 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     1561 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     1621 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     1681 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     1741 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     1801 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     1861 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa
     1921 tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcga cggtatcgat
     1981 aagcttgata tcgaattcct gcagcccggg ggatccacta gttctagagc ggccgccacc
     2041 gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     2101 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2161 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2221 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2281 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2341 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2401 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2461 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2521 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2581 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2641 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2701 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2761 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2821 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2881 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2941 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3001 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3061 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3121 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3181 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3241 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3301 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3361 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3421 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3481 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3541 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3601 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3661 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3721 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3781 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3841 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3901 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3961 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4021 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4081 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4141 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4201 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4261 ctgcaccata aacgacatta ctatatatat aatataggaa gcatttaata gaacagcatc
     4321 gtaatatatg tgtactttgc agttatgacg ccagatggca gtagtggaag atattcttta
     4381 ttgaaaaata gcttgtcacc ttacgtacaa tcttgatccg gagcttttct ttttttgccg
     4441 attaagaatt aattcggtcg aaaaaagaaa aggagagggc caagagggag ggcattggtg
     4501 actattgagc acgtgagtat acgtgattaa gcacacaaag gcagcttgga gtatgtctgt
     4561 tattaatttc acaggtagtt ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga
     4621 ggccgcagaa tgtgctctag attccgatgc tgacttgctg ggtattatat gtgtgcccaa
     4681 tagaaagaga acaattgacc cggttattgc aaggaaaatt tcaagtcttg taaaagcata
     4741 taaaaatagt tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc aacctaagga
     4801 ggatgttttg gctctggtca atgattacgg cattgatatc gtccaactgc atggagatga
     4861 gtcgtggcaa gaataccaag agttcctcgg tttgccagtt attaaaagac tcgtatttcc
     4921 aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt ttattccctt
     4981 gtttgattca gaagcaggtg ggacaggtga acttttggat tggaactcga tttctgactg
     5041 ggttggaagg caagagagcc ccgaaagctt acattttatg ttagctggtg gactgacgcc
     5101 agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg taagcggagg
     5161 tgtggagaca aatggtgtaa aagactctaa caaaatagca aatttcgtca aaaatgctaa
     5221 gaaataggtt attactgagt agtatttatt taagtattgt ttgtgcactt gcctaatcat
     5281 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5341 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5401 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgt
//
