LOCUS       sequence-234421-        5445 bp    DNA     circular SYN 15-APR-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 5445)
  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 5445)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..5445
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             421..1755
                     /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(1860..2315)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1997..2018)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2209..2228
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2445..2467
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2459..2476
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2460..2476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2486..2505
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2486..2504
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2529..2545
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2530..2546
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2580..2596)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2580..2596)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    2627..3130
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(3185..3205)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3185..3203)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3224..3240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3224..3240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3237..3259)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3248..3264
                     /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(3272..3278,3279..3296,3297..3302))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3455..3472)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3626..4214)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3706..3725)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4385..5176,5177..5245))
                     /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     5008..5027
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5246..5350)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5418..5436
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatagcca tcctcatgaa aactgtgtaa cataataacc gaagtgtcga aaaggtggca
      241 ccttgtccaa ttgaacacgc tcgatgaaaa aaataagata tatataaggt taagtaaagc
      301 gtctgttaga aaggaagttt ttcctttttc ttgctctctt gtcttttcat ctactatttc
      361 cttcgtgtaa tacagggtcg tcagatacat agatacaatt ctattacccc catccataca
      421 atgccatctc atttcgatac tgttcaacta cacgccggcc aagagaaccc tggtgacaat
      481 gctcacagat ccagagctgt accaatttac gccaccactt cttatgtttt cgaaaactct
      541 aagcatggtt cgcaattgtt tggtctagaa gttccaggtt acgtctattc ccgtttccaa
      601 aacccaacca gtaatgtttt ggaagaaaga attgctgctt tagaaggtgg tgctgctgct
      661 ttggctgttt cctccggtca agccgctcaa acccttgcca tccaaggttt ggcacacact
      721 ggtgacaaca tcgtttccac ttcttactta tacggtggta cttataacca gttcaaaatc
      781 tcgttcaaaa gatttggtat cgaggctaga tttgttgaag gtgacaatcc agaagaattc
      841 gaaaaggtct ttgatgaaag aaccaaggct gtttatttgg aaaccattgg taatccaaag
      901 tacaatgttc cggattttga aaaaattgtt gcaattgctc acaaacacgg tattccagtt
      961 gtcgttgaca acacatttgg tgccggtggt tacttctgtc agccaattaa atacggtgct
     1021 gatattgtaa cacattctgc taccaaatgg attggtggtc atggtactac tatcggtggt
     1081 attattgttg actctggtaa gttcccatgg aaggactacc cagaaaagtt ccctcaattc
     1141 tctcaacctg ccgaaggata tcacggtact atctacaatg aagcctacgg taacttggca
     1201 tacatcgttc atgttagaac tgaactatta agagatttgg gtccattgat gaacccattt
     1261 gcctctttct tgctactaca aggtgttgaa acattatctt tgagagctga aagacacggt
     1321 gaaaatgcat tgaagttagc caaatggtta gaacaatccc catacgtatc ttgggtttca
     1381 taccctggtt tagcatctca ttctcatcat gaaaatgcta agaagtatct atctaacggt
     1441 ttcggtggtg tcttatcttt cggtgtaaaa gacttaccaa atgccgacaa ggaaactgac
     1501 ccattcaaac tttctggtgc tcaagttgtt gacaatttaa agcttgcctc taacttggcc
     1561 aatgttggtg atgccaagac cttagtcatt gctccatact tcactaccca caaacaatta
     1621 aatgacaaag aaaagttggc atctggtgtt accaaggact taattcgtgt ctctgttggt
     1681 atcgaattta ttgatgacat tattgcagac ttccagcaat cttttgaaac tgttttcgct
     1741 ggccaaaaac catgagtgtg cgtaatgagt tgtaaaatta tgtataaacc tactttctct
     1801 cacaagttat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     1861 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     1921 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     1981 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2041 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2101 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2161 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2221 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2281 acccgccgcg cttaatgcgc cgctacaggg cgcgtcgcgc cattcgccat tcaggctgcg
     2341 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     2401 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     2461 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
     2521 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg
     2581 atccactagt tctagagcgg ccgccctctt agcgcttctt aggacggatc gcttgcctgt
     2641 aacttacacg cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt
     2701 atttattttt atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta
     2761 cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta
     2821 catatatatt tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa
     2881 gtaaaatgta aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa
     2941 ttcggcatta atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc
     3001 cccctagagt cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt
     3061 tactttctat ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat
     3121 agcacgtgat gaaaaggacc caggtggcgc ggccgccacc gcggtggagc tccagctttt
     3181 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     3241 tgtgaaattg ttatccgctc acaattccac acaacatagg agccggaagc ataaagtgta
     3301 aagcctgggg tgcctaatga gtgaggtaac tcacattaat tgcgttgcgc tcactgcccg
     3361 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3421 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3481 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3541 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3601 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3661 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3721 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3781 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3841 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3901 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3961 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4021 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     4081 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4141 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4201 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4261 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4321 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4381 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4441 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4501 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4561 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4621 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4681 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4741 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4801 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4861 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4921 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4981 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5041 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5101 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5161 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5221 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5281 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5341 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     5401 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc
//
