LOCUS       sequence-238644-        4825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1 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 4825)
  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 4825)
  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
FEATURES             Location/Qualifiers
     source          1..4825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..122
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        143..424
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             425..1099
                     /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"
     rep_origin      complement(1196..1651)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1333..1354)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1545..1564
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1777..1799
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1791..1808
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1792..1808
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1818..1837
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1818..1836
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1861..1877
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1862..1878
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1912..1928)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1912..1928)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    1959..2462
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(2517..2537)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2517..2535)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2556..2572)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2556..2572)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2569..2591)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2580..2596
                     /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(2604..2610,2611..2628,2629..2634))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2649..2670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2787..2804)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2958..3546)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3038..3057)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3717..4508,4509..4577))
                     /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     4340..4359
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4578..4682)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4750..4768
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(4806..4825,1..3))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
       61 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
      121 tactgagagt gcaccataga tcaacgacat tactatatat ataatatagg aagcatttaa
      181 tagaacagca tcgtaatata tgtgtacttt gcagttatga cgccagatgg cagtagtgga
      241 agatattctt tattgaaaaa tagcttgtca ccttacgtac aatcttgatc cggagctttt
      301 ctttttttgc cgattaagaa ttaattcggt cgaaaaaaga aaaggagagg gccaagaggg
      361 agggcattgg tgactattga gcacgtgagt atacgtgatt aagcacacaa aggcagcttg
      421 gagtatgtct gttattaatt tcacaggtag ttctggtcca ttggtgaaag tttgcggctt
      481 gcagagcaca gaggccgcag aatgtgctct agattccgat gctgacttgc tgggtattat
      541 atgtgtgccc aatagaaaga gaacaattga cccggttatt gcaaggaaaa tttcaagtct
      601 tgtaaaagca tataaaaata gttcaggcac tccgaaatac ttggttggcg tgtttcgtaa
      661 tcaacctaag gaggatgttt tggctctggt caatgattac ggcattgata tcgtccaact
      721 gcatggagat gagtcgtggc aagaatacca agagttcctc ggtttgccag ttattaaaag
      781 actcgtattt ccaaaagact gcaacatact actcagtgca gcttcacaga aacctcattc
      841 gtttattccc ttgtttgatt cagaagcagg tgggacaggt gaacttttgg attggaactc
      901 gatttctgac tgggttggaa ggcaagagag ccccgaaagc ttacatttta tgttagctgg
      961 tggactgacg ccagaaaatg ttggtgatgc gcttagatta aatggcgtta ttggtgttga
     1021 tgtaagcgga ggtgtggaga caaatggtgt aaaagactct aacaaaatag caaatttcgt
     1081 caaaaatgct aagaaatagg ttattactga gtagtattta tttaattata tcagttatta
     1141 ccctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     1201 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1261 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     1321 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1381 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     1441 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1501 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     1561 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     1621 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     1681 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     1741 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     1801 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc
     1861 tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta
     1921 gttctagagc ggccgccctc ttagcgcttc ttaggacgga tcgcttgcct gtaacttaca
     1981 cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt ttatttattt
     2041 ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt tacggaatga
     2101 agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt tacatatata
     2161 tttattagac aagaaaagca gattaaatag atatacattc gattaacgat aagtaaaatg
     2221 taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac aattcggcat
     2281 taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga tccccctaga
     2341 gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt tttactttct
     2401 atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt atagcacgtg
     2461 atgaaaagga cccaggtggc gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     2521 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2581 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2641 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2701 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2761 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2821 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2881 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2941 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3001 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3061 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3121 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3181 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3241 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3301 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3361 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3421 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3481 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3541 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3601 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3661 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3721 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3781 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3841 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3901 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3961 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4021 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4081 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4141 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4201 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4261 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4321 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4381 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4441 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4501 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4561 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4621 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     4681 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     4741 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     4801 gaaaacctct gacacatgca gctcc
//
