LOCUS       sequence-238646-        4925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  URA3 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 4925)
  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 4925)
  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 URA3
FEATURES             Location/Qualifiers
     source          1..4925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..27)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     127..146
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        170..385
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             386..1189
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1320..1775)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1457..1478)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1669..1688
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1901..1923
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1915..1932
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1916..1932
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1942..1961
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1942..1960
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1985..2001
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1986..2002
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2036..2052)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2036..2052)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    2083..2586
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(2641..2661)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2641..2659)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2680..2696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2680..2696)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2693..2715)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2704..2720
                     /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(2728..2734,2735..2752,2753..2758))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2773..2794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2911..2928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3082..3670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3162..3181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3841..4632,4633..4701))
                     /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     4464..4483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4702..4806)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4874..4892
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
       61 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
      121 actatgcggc atcagagcag attgtactga gagtgcacca taccaccttt tcaattcatc
      181 attttttttt tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat
      241 ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat
      301 atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa
      361 aacctgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac
      421 tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt
      481 gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg
      541 tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg
      601 cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag
      661 aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat
      721 agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg
      781 tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga
      841 attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc
      901 gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga
      961 tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagacgc
     1021 attgggtcaa cagtatagaa ccgtggatga tgtggtctct acaggatctg acattattat
     1081 tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag
     1141 aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt
     1201 ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt
     1261 attaccctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     1321 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     1381 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     1441 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     1501 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     1561 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     1621 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     1681 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     1741 acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac
     1801 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     1861 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     1921 acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc
     1981 cccctcgagg tcgacggtat cgataagctt gatatcgaat tcctgcagcc cgggggatcc
     2041 actagttcta gagcggccgc cctcttagcg cttcttagga cggatcgctt gcctgtaact
     2101 tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt
     2161 atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga
     2221 atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata
     2281 tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa
     2341 aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg
     2401 gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc
     2461 tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact
     2521 ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca
     2581 cgtgatgaaa aggacccagg tggcgcggcc gccaccgcgg tggagctcca gcttttgttc
     2641 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2701 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2761 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2821 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2881 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2941 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3001 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3061 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3121 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3181 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3241 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3301 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3361 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3421 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3481 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3541 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3601 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3661 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3721 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3781 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3841 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3901 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3961 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4021 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4081 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4141 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4201 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4261 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4321 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4381 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4441 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4501 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4561 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4621 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4681 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4741 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4801 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4861 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     4921 cggtg
//
