LOCUS       sequence-238635-        6048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LEU2 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 6048)
  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 6048)
  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 LEU2
FEATURES             Location/Qualifiers
     source          1..6048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        144..551
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             552..1646
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(2140..2159)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2259..2281
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2319..2337)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2405..2509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2510..2578,2579..3370)
                     /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     complement(2728..2747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3541..4129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4030..4049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4283..4300
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4417..4438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4453..4458,4459..4476,4477..4483)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4491..4507
                     /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)."
     primer_bind     4496..4518
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4515..4531
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4515..4531
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4550..4570
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4552..4570
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    4625..5128
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5159..5175
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5159..5175
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5209..5225)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5210..5226)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5250..5269)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5251..5269)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5279..5296)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5279..5295)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5288..5310)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5436..5891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5523..5542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5733..5754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tcaattgtcc tgtacttcct tgttcatgtg tgttcaaaaa cgttatattt ataggataat
       61 tatactctat ttctcaacaa gtaattggtt gtttggccga gcggtctaag gcgcctgatt
      121 caagaaatat cttgaccgca gttaactgtg ggaatactca ggtatcgtaa gatgcaagag
      181 ttcgaatctc ttagcaacca ttattttttt cctcaacata acgagaacac acaggggcgc
      241 tatcgcacag aatcaaattc gatgactgga aattttttgt taatttcaga ggtcgcctga
      301 cgcatatacc tttttcaact gaaaaattgg gagaaaaagg aaaggtgaga gcgccggaac
      361 cggcttttca tatagaatag agaagcgttc atgactaaat gcttgcatca caatacttga
      421 agttgacaat attatttaag gacctattgt tttttccaat aggtggttag caatcgtctt
      481 actttctaac ttttcttacc ttttacattt cagcaatata tatatatata tttcaaggat
      541 ataccattct aatgtctgcc cctaagaaga tcgtcgtttt gccaggtgac cacgttggtc
      601 aagaaatcac agccgaagcc attaaggttc ttaaagctat ttctgatgtt cgttccaatg
      661 tcaagttcga tttcgaaaat catttaattg gtggtgctgc tatcgatgct acaggtgttc
      721 cacttccaga tgaggcgctg gaagcctcca agaaggctga tgccgttttg ttaggtgctg
      781 tgggtggtcc taaatggggt accggtagtg ttagacctga acaaggttta ctaaaaatcc
      841 gtaaagaact tcaattgtac gccaacttaa gaccatgtaa ctttgcatcc gactctcttt
      901 tagacttatc tccaatcaag ccacaatttg ctaaaggtac tgacttcgtt gttgtcagag
      961 aattagtggg aggtatttac tttggtaaga gaaaggaaga cgatggtgat ggtgtcgctt
     1021 gggatagtga acaatacacc gttccagaag tgcaaagaat cacaagaatg gccgctttca
     1081 tggccctaca acatgagcca ccattgccta tttggtcctt ggataaagct aatgttttgg
     1141 cctcttcaag attatggaga aaaactgtgg aggaaaccat caagaacgaa ttccctacat
     1201 tgaaggttca acatcaattg attgattctg ccgccatgat cctagttaag aacccaaccc
     1261 acctaaatgg tattataatc accagcaaca tgtttggtga tatcatctcc gatgaagcct
     1321 ccgttatccc aggttccttg ggtttgttgc catctgcgtc cttggcctct ttgccagaca
     1381 agaacaccgc atttggtttg tacgaaccat gccacggttc tgctccagat ttgccaaaga
     1441 ataaggtcaa ccctatcgcc actatcttgt ctgctgcaat gatgttgaaa ttgtcattga
     1501 acttgcctga agaaggtaag gccattgaag atgcagttaa aaaggttttg gatgcaggta
     1561 tcagaactgg tgatttaggt ggttccaaca gtaccaccga agtcggtgat gctgtcgccg
     1621 aagaagttaa gaaaatcctt gcttaaaaag attctctttt tttatgatat ttgtacataa
     1681 actttataaa tgaaattcat aatagaaacg acacgaaatt acaaaatgga atatgttcat
     1741 agggtagacg aaactatata cgcaatctac atacatttat caagaaggag aaaaaggagg
     1801 atgtaaagga atacaggtaa gcaaattgat actaatggct caacgtgata aggaaaaaga
     1861 attgcacttt aacattaata ttgacaagga ggagggcacc acacaaaaag ttaggtgtaa
     1921 cagaaaatca tgaaactatg attcctaatt tatatattgg aggattttct ctaaaaaaaa
     1981 aaaaatacaa caaataaaaa acactcaatg acctgaccat ttgatggagt ttaagtcaat
     2041 accttcttga accatttccc ataatggtga aagttccctc aagaatttta ctctgtcaga
     2101 aacggcctta acgacgtagt cgatatggtg cactctcagt acaatctgct ctgatgccgc
     2161 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     2221 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     2281 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     2341 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     2401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2461 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2521 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2581 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2641 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2701 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2761 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2821 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2881 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2941 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3001 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3061 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3121 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3181 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3241 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3301 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3361 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3421 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3481 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3541 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3601 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3661 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3721 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3781 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3841 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3901 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3961 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4021 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4081 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4141 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     4201 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     4261 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     4321 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     4381 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     4441 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     4501 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     4561 tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgcgcca cctgggtcct
     4621 tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt
     4681 aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta
     4741 aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag
     4801 gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg
     4861 attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct
     4921 aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt
     4981 ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa
     5041 acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga
     5101 ggcgcgtgta agttacaggc aagcgatccg tcctaagaag cgctaagagg gcggccgctc
     5161 tagaactagt ggatcccccg ggctgcagga attcgatatc aagcttatcg ataccgtcga
     5221 cctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     5281 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     5341 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     5401 cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag
     5461 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5521 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5581 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5641 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5701 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5761 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5821 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5881 gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     5941 atatcgaccg gtcgaggaga acttctagta tatctacata cctaatatta ttgccttatt
     6001 aaaaatggaa tcccaacaat tacatcaaaa tccacattct cttcaaaa
//
