LOCUS       sequence-238634-        5008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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 5008)
  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 5008)
  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 HIS3
FEATURES             Location/Qualifiers
     source          1..5008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..47
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..381
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             382..1044
                     /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(1304..1759)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1441..1462)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1653..1672
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1885..1907
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1899..1916
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1900..1916
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1926..1945
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1926..1944
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1969..1985
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1970..1986
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2020..2036)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2020..2036)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    2067..2570
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(2625..2645)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2625..2643)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2664..2680)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2664..2680)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2677..2699)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2688..2704
                     /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(2712..2718,2719..2736,2737..2742))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2757..2778
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2895..2912)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3066..3654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3146..3165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3825..4616,4617..4685))
                     /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     4448..4467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4686..4790)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4858..4876
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4914..4936)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
       61 ataaattccc gttttaagag cttggtgagc gctaggagtc actgccaggt atcgtttgaa
      121 cacggcatta gtcagggaag tcataacaca gtcctttccc gcaattttct ttttctatta
      181 ctcttggcct cctctagtac actctatatt tttttatgcc tcggtaatga ttttcatttt
      241 tttttttcca cctagcggat gactcttttt ttttcttagc gattggcatt atcacataat
      301 gaattataca ttatataaag taatgtgatt tcttcgaaga atatactaaa aaatgagcag
      361 gcaagataaa cgaaggcaaa gatgacagag cagaaagccc tagtaaagcg tattacaaat
      421 gaaaccaaga ttcagattgc gatctcttta aagggtggtc ccctagcgat agagcactcg
      481 atcttcccag aaaaagaggc agaagcagta gcagaacagg ccacacaatc gcaagtgatt
      541 aacgtccaca caggtatagg gtttctggac catatgatac atgctctggc caagcattcc
      601 ggctggtcgc taatcgttga gtgcattggt gacttacaca tagacgacca tcacaccact
      661 gaagactgcg ggattgctct cggtcaagct tttaaagagg ccctaggggc cgtgcgtgga
      721 gtaaaaaggt ttggatcagg atttgcgcct ttggatgagg cactttccag agcggtggta
      781 gatctttcga acaggccgta cgcagttgtc gaacttggtt tgcaaaggga gaaagtagga
      841 gatctctctt gcgagatgat cccgcatttt cttgaaagct ttgcagaggc tagcagaatt
      901 accctccacg ttgattgtct gcgaggcaag aatgatcatc accgtagtga gagtgcgttc
      961 aaggctcttg cggttgccat aagagaagcc acctcgccca atggtaccaa cgatgttccc
     1021 tccaccaaag gtgttcttat gtagtgacac cgattattta aagctgcagc atacgatata
     1081 tatacatgtg tatatatgta tacctatgaa tgtcagtaag tatgtatacg aacagtatga
     1141 tactgaagat gacaaggtaa tgcatcattc tatacgtgtc attctgaacg aggcgcgctt
     1201 tccttttttc tttttgcttt ttcttttttt ttctcttgaa ctcgacggat ctatgcggtg
     1261 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat
     1321 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     1381 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     1441 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     1501 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     1561 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     1621 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     1681 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     1741 gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     1801 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     1861 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgagc
     1921 gcgcgtaata cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcgacg
     1981 gtatcgataa gcttgatatc gaattcctgc agcccggggg atccactagt tctagagcgg
     2041 ccgccctctt agcgcttctt aggacggatc gcttgcctgt aacttacacg cgcctcgtat
     2101 cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt atgttttgta
     2161 tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat
     2221 aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa
     2281 gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg
     2341 attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga
     2401 gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc
     2461 ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat ttttaattta
     2521 tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat gaaaaggacc
     2581 caggtggcgc ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta
     2641 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2701 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2761 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2821 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2881 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2941 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3001 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3061 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3781 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3841 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3901 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3961 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4021 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4081 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4141 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4201 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4261 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4321 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4381 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4441 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4501 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4561 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4621 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4681 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4741 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4801 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4861 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4921 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4981 gcccgtcagg gcgcgtcagc gggtgttg
//
