LOCUS       sequence-238639-        5441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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 5441)
  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 5441)
  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 MET17
FEATURES             Location/Qualifiers
     source          1..5441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..45
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(83..105)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     205..224
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             475..1809
                     /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"
     rep_origin      complement(1914..2369)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2051..2072)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2263..2282
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2495..2517
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2509..2526
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2510..2526
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2536..2555
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2536..2554
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2579..2595
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2580..2596
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2630..2646)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2630..2646)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    2677..3180
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(3235..3255)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3235..3253)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3274..3290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3274..3290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3287..3309)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3298..3314
                     /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(3322..3328,3329..3346,3347..3352))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3367..3388
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3505..3522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3676..4264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3756..3775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4435..5226,5227..5295))
                     /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     5058..5077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5296..5400)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
       61 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
      121 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
      181 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      241 gccatcctca tgaaaactgt gtaacataat aaccgaagtg tcgaaaaggt ggcaccttgt
      301 ccaattgaac acgctcgatg aaaaaaataa gatatatata aggttaagta aagcgtctgt
      361 tagaaaggaa gtttttcctt tttcttgctc tcttgtcttt tcatctacta tttccttcgt
      421 gtaatacagg gtcgtcagat acatagatac aattctatta cccccatcca tacaatgcca
      481 tctcatttcg atactgttca actacacgcc ggccaagaga accctggtga caatgctcac
      541 agatccagag ctgtaccaat ttacgccacc acttcttatg ttttcgaaaa ctctaagcat
      601 ggttcgcaat tgtttggtct agaagttcca ggttacgtct attcccgttt ccaaaaccca
      661 accagtaatg ttttggaaga aagaattgct gctttagaag gtggtgctgc tgctttggct
      721 gtttcctccg gtcaagccgc tcaaaccctt gccatccaag gtttggcaca cactggtgac
      781 aacatcgttt ccacttctta cttatacggt ggtacttata accagttcaa aatctcgttc
      841 aaaagatttg gtatcgaggc tagatttgtt gaaggtgaca atccagaaga attcgaaaag
      901 gtctttgatg aaagaaccaa ggctgtttat ttggaaacca ttggtaatcc aaagtacaat
      961 gttccggatt ttgaaaaaat tgttgcaatt gctcacaaac acggtattcc agttgtcgtt
     1021 gacaacacat ttggtgccgg tggttacttc tgtcagccaa ttaaatacgg tgctgatatt
     1081 gtaacacatt ctgctaccaa atggattggt ggtcatggta ctactatcgg tggtattatt
     1141 gttgactctg gtaagttccc atggaaggac tacccagaaa agttccctca attctctcaa
     1201 cctgccgaag gatatcacgg tactatctac aatgaagcct acggtaactt ggcatacatc
     1261 gttcatgtta gaactgaact attaagagat ttgggtccat tgatgaaccc atttgcctct
     1321 ttcttgctac tacaaggtgt tgaaacatta tctttgagag ctgaaagaca cggtgaaaat
     1381 gcattgaagt tagccaaatg gttagaacaa tccccatacg tatcttgggt ttcataccct
     1441 ggtttagcat ctcattctca tcatgaaaat gctaagaagt atctatctaa cggtttcggt
     1501 ggtgtcttat ctttcggtgt aaaagactta ccaaatgccg acaaggaaac tgacccattc
     1561 aaactttctg gtgctcaagt tgttgacaat ttaaagcttg cctctaactt ggccaatgtt
     1621 ggtgatgcca agaccttagt cattgctcca tacttcacta cccacaaaca attaaatgac
     1681 aaagaaaagt tggcatctgg tgttaccaag gacttaattc gtgtctctgt tggtatcgaa
     1741 tttattgatg acattattgc agacttccag caatcttttg aaactgtttt cgctggccaa
     1801 aaaccatgag tgtgcgtaat gagttgtaaa attatgtata aaccgacttt ctctcacaag
     1861 ttatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     1921 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     1981 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2041 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2101 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2161 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2221 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2281 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2341 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     2401 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     2461 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     2521 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg gccccccctc
     2581 gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg atccactagt
     2641 tctagagcgg ccgccctctt agcgcttctt aggacggatc gcttgcctgt aacttacacg
     2701 cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt
     2761 atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag
     2821 aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt
     2881 tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta
     2941 aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta
     3001 atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt
     3061 cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat
     3121 ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat
     3181 gaaaaggacc caggtggcgc ggccgccacc gcggtggagc tccagctttt gttcccttta
     3241 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3301 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3361 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3421 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     3481 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3541 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3601 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3661 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3721 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3781 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3841 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3901 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3961 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4021 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4081 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4141 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4201 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4261 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4321 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4381 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4441 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4501 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4561 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4621 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4681 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4741 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4801 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4861 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4921 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4981 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5041 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5101 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5161 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5221 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5281 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5341 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5401 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc a
//
