LOCUS       sequence-238926-        6007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-HIS3 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 6007)
  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 6007)
  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-HIS3
FEATURES             Location/Qualifiers
     source          1..6007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     24..43
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        190..377
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             378..1040
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1300..1755)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1437..1458)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1649..1668
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1881..1903
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1895..1912
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1896..1912
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1922..1941
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1922..1940
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1965..1981
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1966..1982
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2016..2032)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2016..2032)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    2063..2566
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(2621..2641)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2621..2639)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2660..2676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2660..2676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2673..2695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2684..2700
                     /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(2708..2714,2715..2732,2733..2738))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2753..2774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2891..2908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3062..3650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3142..3161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3821..4612,4613..4681))
                     /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     4444..4463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4682..4786)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4827..5108
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             5109..5783
                     /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"
     primer_bind     5853..5871
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5909..5931)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa
       61 attcccgttt taagagcttg gtgagcgcta ggagtcactg ccaggtatcg tttgaacacg
      121 gcattagtca gggaagtcat aacacagtcc tttcccgcaa ttttcttttt ctattactct
      181 tggcctcctc tagtacactc tatatttttt tatgcctcgg taatgatttt catttttttt
      241 tttccaccta gcggatgact cttttttttt cttagcgatt ggcattatca cataatgaat
      301 tatacattat ataaagtaat gtgatttctt cgaagaatat actaaaaaat gagcaggcaa
      361 gataaacgaa ggcaaagatg acagagcaga aagccctagt aaagcgtatt acaaatgaaa
      421 ccaagattca gattgcgatc tctttaaagg gtggtcccct agcgatagag cactcgatct
      481 tcccagaaaa agaggcagaa gcagtagcag aacaggccac acaatcgcaa gtgattaacg
      541 tccacacagg tatagggttt ctggaccata tgatacatgc tctggccaag cattccggct
      601 ggtcgctaat cgttgagtgc attggtgact tacacataga cgaccatcac accactgaag
      661 actgcgggat tgctctcggt caagctttta aagaggccct aggggccgtg cgtggagtaa
      721 aaaggtttgg atcaggattt gcgcctttgg atgaggcact ttccagagcg gtggtagatc
      781 tttcgaacag gccgtacgca gttgtcgaac ttggtttgca aagggagaaa gtaggagatc
      841 tctcttgcga gatgatcccg cattttcttg aaagctttgc agaggctagc agaattaccc
      901 tccacgttga ttgtctgcga ggcaagaatg atcatcaccg tagtgagagt gcgttcaagg
      961 ctcttgcggt tgccataaga gaagccacct cgcccaatgg taccaacgat gttccctcca
     1021 ccaaaggtgt tcttatgtag tgacaccgat tatttaaagc tgcagcatac gatatatata
     1081 catgtgtata tatgtatacc tatgaatgtc agtaagtatg tatacgaaca gtatgatact
     1141 gaagatgaca aggtaatgca tcattctata cgtgtcattc tgaacgaggc gcgctttcct
     1201 tttttctttt tgctttttct ttttttttct cttgaactcg acggatctat gcggtgtgaa
     1261 ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt
     1321 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     1381 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     1441 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     1501 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     1561 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     1621 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     1681 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     1741 cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     1801 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     1861 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc
     1921 gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg tcgacggtat
     1981 cgataagctt gatatcgaat tcctgcagcc cgggggatcc actagttcta gagcggccgc
     2041 cctcttagcg cttcttagga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt
     2101 taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg
     2161 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac
     2221 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa
     2281 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt
     2341 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag
     2401 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg
     2461 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata
     2521 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg
     2581 tggcgcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg
     2641 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2701 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2761 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2821 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2881 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2941 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3001 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3061 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3781 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4741 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4801 aagtgccacc tgacgtctgc accataaacg acattactat atatataata taggaagcat
     4861 ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag
     4921 tggaagatat tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc
     4981 ttttcttttt ttgccgatta agaattaatt cggtcgaaaa aagaaaagga gagggccaag
     5041 agggagggca ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag
     5101 cttggagtat gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg
     5161 gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta
     5221 ttatatgtgt gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa
     5281 gtcttgtaaa agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc
     5341 gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc
     5401 aactgcatgg agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta
     5461 aaagactcgt atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc
     5521 attcgtttat tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga
     5581 actcgatttc tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag
     5641 ctggtggact gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg
     5701 ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt
     5761 tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt
     5821 gcacttgcct aatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     5881 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     5941 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     6001 ttggcgg
//
