LOCUS       sequence-239160-        6239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-KanMx 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 6239)
  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 6239)
  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-KanMx
FEATURES             Location/Qualifiers
     source          1..6239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     67..86
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        107..388
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             389..1063
                     /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(1160..1615)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1297..1318)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1509..1528
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1741..1763
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1755..1772
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1756..1772
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1782..1801
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1782..1800
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1825..1841
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1826..1842
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1876..1892)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1876..1892)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    1923..2426
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(2481..2501)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2481..2499)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2520..2536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2520..2536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2533..2555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2544..2560
                     /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(2568..2574,2575..2592,2593..2598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2613..2634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2751..2768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2922..3510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3002..3021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3681..4472,4473..4541))
                     /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     4304..4323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4542..4646)
                     /gene="bla"
                     /label=AmpR promoter
     gene            4719..6075
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        4719..5062
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5063..5872
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5130..5149)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      5878..6075
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     6128..6146
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6184..6206)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
       61 actatgcggc atcagagcag attgtactga gagtgcacca tagatcaacg acattactat
      121 atatataata taggaagcat ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt
      181 atgacgccag atggcagtag tggaagatat tctttattga aaaatagctt gtcaccttac
      241 gtacaatctt gatccggagc ttttcttttt ttgccgatta agaattaatt cggtcgaaaa
      301 aagaaaagga gagggccaag agggagggca ttggtgacta ttgagcacgt gagtatacgt
      361 gattaagcac acaaaggcag cttggagtat gtctgttatt aatttcacag gtagttctgg
      421 tccattggtg aaagtttgcg gcttgcagag cacagaggcc gcagaatgtg ctctagattc
      481 cgatgctgac ttgctgggta ttatatgtgt gcccaataga aagagaacaa ttgacccggt
      541 tattgcaagg aaaatttcaa gtcttgtaaa agcatataaa aatagttcag gcactccgaa
      601 atacttggtt ggcgtgtttc gtaatcaacc taaggaggat gttttggctc tggtcaatga
      661 ttacggcatt gatatcgtcc aactgcatgg agatgagtcg tggcaagaat accaagagtt
      721 cctcggtttg ccagttatta aaagactcgt atttccaaaa gactgcaaca tactactcag
      781 tgcagcttca cagaaacctc attcgtttat tcccttgttt gattcagaag caggtgggac
      841 aggtgaactt ttggattgga actcgatttc tgactgggtt ggaaggcaag agagccccga
      901 aagcttacat tttatgttag ctggtggact gacgccagaa aatgttggtg atgcgcttag
      961 attaaatggc gttattggtg ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga
     1021 ctctaacaaa atagcaaatt tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta
     1081 tttatttaat tatatcagtt attaccctat gcggtgtgaa ataccgcaca gatgcgtaag
     1141 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     1201 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     1261 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     1321 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     1381 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     1441 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     1501 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     1561 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     1621 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     1681 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     1741 cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     1801 gcgaattggg taccgggccc cccctcgagg tcgacggtat cgataagctt gatatcgaat
     1861 tcctgcagcc cgggggatcc actagttcta gagcggccgc cctcttagcg cttcttagga
     1921 cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg
     1981 aatttactct gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa
     2041 gaaggtagaa gagttacgga atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa
     2101 caaaaagcgt actttacata tatatttatt agacaagaaa agcagattaa atagatatac
     2161 attcgattaa cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac
     2221 agacaagatg aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta
     2281 gtatttgttg gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatttttt
     2341 ctttaatttc tttttttact ttctattttt aatttatata tttatattaa aaaatttaaa
     2401 ttataattat ttttatagca cgtgatgaaa aggacccagg tggcgcggcc gccaccgcgg
     2461 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     2521 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     2581 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     2641 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     2701 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2761 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2821 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3361 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3601 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3661 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3721 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3781 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3841 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3901 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3961 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4021 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4081 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4141 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4201 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4261 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4321 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4381 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4441 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4501 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4561 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4621 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcttg
     4681 tttagcttgc ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc
     4741 tccttgacag tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt
     4801 agcccataca tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc
     4861 gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag
     4921 acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc
     4981 cactgaggtt cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca
     5041 tcacatccga acataaacaa ccatgggtaa ggaaaagact cacgtttcga ggccgcgatt
     5101 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     5161 atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa
     5221 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     5281 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     5341 gttactcacc actgcgatcc ccggcaaaac agcattccag gtattagaag aatatcctga
     5401 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     5461 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     5521 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     5581 tgaacaagtc tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac
     5641 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     5701 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     5761 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     5821 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagtact
     5881 gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta
     5941 gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc
     6001 tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg
     6061 ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagatca tgacattaac
     6121 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     6181 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccg
//
