LOCUS       sequence-238929-        6440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-MET17 ICE plasmid used to facilitate yeast recombination 
            cloning and stable genomic Integration after CEN Excision..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6440)
  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 6440)
  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: pICE TRP1-MET17
FEATURES             Location/Qualifiers
     source          1..6440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..21)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     121..143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(181..199)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(269..943)
                     /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"
     promoter        complement(944..1225)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     promoter        1266..1370
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1371..1439,1440..2231)
                     /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     complement(1589..1608)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2402..2990
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2891..2910
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3144..3161
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3278..3299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3314..3319,3320..3337,3338..3344)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3352..3368
                     /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)."
     primer_bind     3357..3379
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3376..3392
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3376..3392
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3411..3431
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3413..3431
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    3486..3989
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4020..4036
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4020..4036
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4070..4086)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4071..4087)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4111..4130)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4112..4130)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4140..4157)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4140..4156)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4149..4171)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4297..4752
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4384..4403)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4594..4615
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4857..6191)
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHGTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENAKKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
ORIGIN
        1 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
       61 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
      121 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
      181 gcctcgtgat acgcctattt ttataggtta atgtcatgat taggcaagtg cacaaacaat
      241 acttaaataa atactactca gtaataacct atttcttagc atttttgacg aaatttgcta
      301 ttttgttaga gtcttttaca ccatttgtct ccacacctcc gcttacatca acaccaataa
      361 cgccatttaa tctaagcgca tcaccaacat tttctggcgt cagtccacca gctaacataa
      421 aatgtaagct ttcggggctc tcttgccttc caacccagtc agaaatcgag ttccaatcca
      481 aaagttcacc tgtcccacct gcttctgaat caaacaaggg aataaacgaa tgaggtttct
      541 gtgaagctgc actgagtagt atgttgcagt cttttggaaa tacgagtctt ttaataactg
      601 gcaaaccgag gaactcttgg tattcttgcc acgactcatc tccatgcagt tggacgatat
      661 caatgccgta atcattgacc agagccaaaa catcctcctt aggttgatta cgaaacacgc
      721 caaccaagta tttcggagtg cctgaactat ttttatatgc ttttacaaga cttgaaattt
      781 tccttgcaat aaccgggtca attgttctct ttctattggg cacacatata atacccagca
      841 agtcagcatc ggaatctaga gcacattctg cggcctctgt gctctgcaag ccgcaaactt
      901 tcaccaatgg accagaacta cctgtgaaat taataacaga catactccaa gctgcctttg
      961 tgtgcttaat cacgtatact cacgtgctca atagtcacca atgccctccc tcttggccct
     1021 ctccttttct tttttcgacc gaattaattc ttaatcggca aaaaaagaaa agctccggat
     1081 caagattgta cgtaaggtga caagctattt ttcaataaag aatatcttcc actactgcca
     1141 tctggcgtca taactgcaaa gtacacatat attacgatgc tgttctatta aatgcttcct
     1201 atattatata tatagtaatg tcgtttatgg tgcagacgtc aggtggcact tttcggggaa
     1261 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1321 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1381 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1441 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1501 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1561 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1621 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1681 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1741 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1801 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1861 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     1921 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1981 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2041 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2101 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2161 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2221 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2281 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2341 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2401 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2461 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2521 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2581 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2641 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2701 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2761 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2821 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2881 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2941 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3001 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3061 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3121 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     3181 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     3241 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     3301 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     3361 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     3421 ctcactaaag ggaacaaaag ctggagctcc accgcggtgg cggccgcgcc acctgggtcc
     3481 ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa atatataaat
     3541 taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt
     3601 aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca
     3661 ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt
     3721 gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc
     3781 taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt
     3841 tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa
     3901 aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg
     3961 aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa gcgctaagag ggcggccgct
     4021 ctagaactag tggatccccc gggctgcagg aattcgatat caagcttatc gataccgtcg
     4081 acctcgaggg ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     4141 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     4201 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     4261 ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa
     4321 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     4381 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4441 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4501 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4561 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4621 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4681 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4741 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4801 cataacttgt gagagaaagt cggtttatac ataattttac aactcattac gcacactcat
     4861 ggtttttggc cagcgaaaac agtttcaaaa gattgctgga agtctgcaat aatgtcatca
     4921 ataaattcga taccaacaga gacacgaatt aagtccttgg taacaccaga tgccaacttt
     4981 tctttgtcat ttaattgttt gtgggtagtg aagtatggag caatgactaa ggtcttggca
     5041 tcaccaacat tggccaagtt agaggcaagc tttaaattgt caacaacttg agcaccagaa
     5101 agtttgaatg ggtcagtttc cttgtcggca tttggtaagt cttttacacc gaaagataag
     5161 acaccaccga aaccgttaga tagatacttc ttagcatttt catgatgaga atgagatgct
     5221 aaaccagggt atgaaaccca agatacgtat ggggattgtt ctaaccattt ggctaacttc
     5281 aatgcatttt caccgtgtct ttcagctctc aaagataatg tttcaacacc ttgtagtagc
     5341 aagaaagagg caaatgggtt catcaatgga cccaaatctc ttaatagttc agttctaaca
     5401 tgaacgatgt atgccaagtt accgtaggct tcattgtaga tagtaccgtg atatccttcg
     5461 gcaggttgag agaattgagg gaacttttct gggtagtcct tccatgggaa cttaccagag
     5521 tcaacaataa taccaccgat agtagtacca tgaccaccaa tccatttggt agcagaatgt
     5581 gttacaatat cagcaccgta tttaattggc tgacagaagt aaccaccggc accaaatgtg
     5641 ttgtcaacga caactggaat accgtgtttg tgagcaattg caacaatttt ttcaaaatcc
     5701 ggaacattgt actttggatt accaatggtt tccaaataaa cagccttggt tctttcatca
     5761 aagacctttt cgaattcttc tggattgtca ccttcaacaa atctagcctc gataccaaat
     5821 cttttgaacg agattttgaa ctggttataa gtaccaccgt ataagtaaga agtggaaacg
     5881 atgttgtcac cagtgtgtgc caaaccttgg atggcaaggg tttgagcggc ttgaccggag
     5941 gaaacagcca aagcagcagc accaccttct aaagcagcaa ttctttcttc caaaacatta
     6001 ctggttgggt tttggaaacg ggaatagacg taacctggaa cttctagacc aaacaattgc
     6061 gaaccatgct tagagttttc gaaaacataa gaagtggtgg cgtaaattgg tacagctctg
     6121 gatctgtgag cattgtcacc agggttctct tggccggcgt gtagttgaac agtatcgaaa
     6181 tgagatggca ttgtatggat gggggtaata gaattgtatc tatgtatctg acgaccctgt
     6241 attacacgaa ggaaatagta gatgaaaaga caagagagca agaaaaagga aaaacttcct
     6301 ttctaacaga cgctttactt aaccttatat atatcttatt tttttcatcg agcgtgttca
     6361 attggacaag gtgccacctt ttcgacactt cggttattat gttacacagt tttcatgagg
     6421 atggctatgg tgcactctca
//
