LOCUS       sequence-239077-        6495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-LEU2 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 6495)
  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 6495)
  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-LEU2
FEATURES             Location/Qualifiers
     source          1..6495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(414..1508)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1509..1916)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2217..2672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2354..2375)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2566..2585
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2649..3029)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2798..2820
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2812..2829
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2813..2829
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2839..2858
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2839..2857
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2866..2973
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2882..2898
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2883..2899
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2933..2949)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2933..2949)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2986..3006)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2986..3004)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3025..3041)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3025..3041)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3038..3060)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3049..3065
                     /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(3073..3079,3080..3097,3098..3103))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3118..3139
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3256..3273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3427..4015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3507..3526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4186..4977,4978..5046))
                     /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     4809..4828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5047..5151)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5192..5473
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5474..6148
                     /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     6218..6236
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6274..6296)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6396..6415
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc attgagtgtt
       61 ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa attaggaatc
      121 atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct ccttgtcaat
      181 attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta tcaatttgct
      241 tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg tagattgcgt
      301 atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc gtttctatta
      361 tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc tttttaagca
      421 aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact gttggaacca
      481 cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc ttcaatggcc
      541 ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga caagatagtg
      601 gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca tggttcgtac
      661 aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg caacaaaccc
      721 aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat gttgctggtg
      781 attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc agaatcaatc
      841 aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc cacagttttt
      901 ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat aggcaatggt
      961 ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac ttctggaacg
     1021 gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct cttaccaaag
     1081 taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc aaattgtggc
     1141 ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct taagttggcg
     1201 tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac actaccggta
     1261 ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt ggaggcttcc
     1321 agcgcctcat ctggaagtgg aacacctgta gcatcgatag cagcaccacc aattaaatga
     1381 ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag aaccttaatg
     1441 gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat cttcttaggg
     1501 gcagacatta gaatggtata tccttgaaat atatatatat atattgctga aatgtaaaag
     1561 gtaagaaaag ttagaaagta agacgattgc taaccaccta ttggaaaaaa caataggtcc
     1621 ttaaataata ttgtcaactt caagtattgt gatgcaagca tttagtcatg aacgcttctc
     1681 tattctatat gaaaagccgg ttccggcgct ctcacctttc ctttttctcc caatttttca
     1741 gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa caaaaaattt ccagtcatcg
     1801 aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt atgttgagga aaaaaataat
     1861 ggttgctaag agattcgaac tcttgcatct tacgatacct gagtattccc acagttaact
     1921 gcggtcaaga tatttcttga atcaggcgcc ttagaccgct cggccaaaca accaattact
     1981 tgttgagaaa tagagtataa ttatcctata aatataacgt ttttgaacac acatgaacaa
     2041 ggaagtacag gacaattgat tttgaagaga atgtggattt tgatgtaatt gttgggattc
     2101 catttttaat aaggcaataa tattaggtat gtagatatac tagaagttct cctcgaccgg
     2161 tcgatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     2221 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2281 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2341 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2401 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2461 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2521 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2581 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2641 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     2701 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     2761 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     2821 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
     2881 ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact
     2941 agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     3001 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3061 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3121 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3181 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     3901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5161 ccgaaaagtg ccacctgacg tctgcaccat aaacgacatt actatatata taatatagga
     5221 agcatttaat agaacagcat cgtaatatat gtgtactttg cagttatgac gccagatggc
     5281 agtagtggaa gatattcttt attgaaaaat agcttgtcac cttacgtaca atcttgatcc
     5341 ggagcttttc tttttttgcc gattaagaat taattcggtc gaaaaaagaa aaggagaggg
     5401 ccaagaggga gggcattggt gactattgag cacgtgagta tacgtgatta agcacacaaa
     5461 ggcagcttgg agtatgtctg ttattaattt cacaggtagt tctggtccat tggtgaaagt
     5521 ttgcggcttg cagagcacag aggccgcaga atgtgctcta gattccgatg ctgacttgct
     5581 gggtattata tgtgtgccca atagaaagag aacaattgac ccggttattg caaggaaaat
     5641 ttcaagtctt gtaaaagcat ataaaaatag ttcaggcact ccgaaatact tggttggcgt
     5701 gtttcgtaat caacctaagg aggatgtttt ggctctggtc aatgattacg gcattgatat
     5761 cgtccaactg catggagatg agtcgtggca agaataccaa gagttcctcg gtttgccagt
     5821 tattaaaaga ctcgtatttc caaaagactg caacatacta ctcagtgcag cttcacagaa
     5881 acctcattcg tttattccct tgtttgattc agaagcaggt gggacaggtg aacttttgga
     5941 ttggaactcg atttctgact gggttggaag gcaagagagc cccgaaagct tacattttat
     6001 gttagctggt ggactgacgc cagaaaatgt tggtgatgcg cttagattaa atggcgttat
     6061 tggtgttgat gtaagcggag gtgtggagac aaatggtgta aaagactcta acaaaatagc
     6121 aaatttcgtc aaaaatgcta agaaataggt tattactgag tagtatttat ttaagtattg
     6181 tttgtgcact tgcctaatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6241 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     6301 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6361 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     6421 agtgcaccat atcgactacg tcgttaaggc cgtttctgac agagtaaaat tcttgaggga
     6481 actttcacca ttatg
//
