LOCUS       sequence-238967-        6722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-HIS3 integrating (YIp) vector for stable genomic integration in
            the BY4741 background..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6722)
  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 6722)
  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: YIp ADE2-HIS3
FEATURES             Location/Qualifiers
     source          1..6722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..20
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(58..80)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     180..199
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        390..765
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             766..2481
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     rep_origin      complement(2686..3141)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2823..2844)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3035..3054
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(3118..3498)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3267..3289
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3281..3298
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3282..3298
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3308..3327
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3308..3326
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3335..3442
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3351..3367
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3352..3368
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3402..3418)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3402..3418)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3455..3475)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3455..3473)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3494..3510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3494..3510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3507..3529)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3518..3534
                     /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(3542..3548,3549..3566,3567..3572))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3587..3608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3725..3742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3896..4484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3976..3995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4655..5446,5447..5515))
                     /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     5278..5297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5516..5620)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5702..5889
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             5890..6552
                     /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"
ORIGIN
        1 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
       61 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
      121 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
      181 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
      241 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
      301 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
      361 tataaatcaa aagaatagat ctgaattaat tcttgaataa tacataactt ttcttaaaag
      421 aatcaaagac agataaaatt taagagatat taaatattag tgagaagccg agaattttgt
      481 aacaccaaca taacactgac atctttaaca acttttaatt atgatacatt tcttacgtca
      541 tgattgatta ttacagctat gctgacaaat gactcttgtt gcatggctac gaaccgggta
      601 atactaagtg attgactctt gctgaccttt tattaagaac taaatggaca atattatgga
      661 gcatttcatg tataaattgg tgcgtaaaat cgttggatct ctcttctaag tacatcctac
      721 tataacaatc aagaaaaaca agaaaatcgg acaaaacaat caagtatgga ttctagaaca
      781 gttggtatat taggaggggg acaattggga cgtatgattg ttgaggcagc aaacaggctc
      841 aacattaaga cggtaatact agatgctgaa aattctcctg ccaaacaaat aagcaactcc
      901 aatgaccacg ttaatggctc cttttccaat cctcttgata tcgaaaaact agctgaaaaa
      961 tgtgatgtgc taacgattga gattgagcat gttgatgttc ctacactaaa gaatcttcaa
     1021 gtaaaacatc ccaaattaaa aatttaccct tctccagaaa caatcagatt gatacaagac
     1081 aaatatattc aaaaagagca tttaatcaaa aatggtatag cagttaccca aagtgttcct
     1141 gtggaacaag ccagtgagac gtccctattg aatgttggaa gagatttggg ttttccattc
     1201 gtcttgaagt cgaggacttt ggcatacgat ggaagaggta acttcgttgt aaagaataag
     1261 gaaatgattc cggaagcttt ggaagtactg aaggatcgtc ctttgtacgc cgaaaaatgg
     1321 gcaccattta ctaaagaatt agcagtcatg attgtgaggt ctgttaacgg tttagtgttt
     1381 tcttacccaa ttgtagagac tatccacaag gacaatattt gtgacttatg ttatgcgcct
     1441 gctagagttc cggactccgt tcaacttaag gcgaagttgt tggcagaaaa tgcaatcaaa
     1501 tcttttcccg gttgtggtat atttggtgtg gaaatgttct atttagaaac aggggaattg
     1561 cttattaacg aaattgcccc aaggcctcac aactctggac attataccat tgatgcttgc
     1621 gtcacttctc aatttgaagc tcatttgaga tcaatattgg atttgccaat gccaaagaat
     1681 ttcacatctt tctccaccat tacaacgaac gccattatgc taaatgttct tggagacaaa
     1741 catacaaaag ataaagagct agaaacttgc gaaagagcat tggcgactcc aggttcctca
     1801 gtgtacttat atggaaaaga gtctagacct aacagaaaag taggtcacat aaatattatt
     1861 gcctccagta tggcggaatg tgaacaaagg ctgaactaca ttacaggtag aactgatatt
     1921 ccaatcaaaa tctctgtcgc tcaaaagttg gacttggaag caatggtcaa accattggtt
     1981 ggaatcatca tgggatcaga ctctgacttg ccggtaatgt ctgccgcatg tgcggtttta
     2041 aaagattttg gcgttccatt tgaagtgaca atagtctctg ctcatagaac tccacatagg
     2101 atgtcagcat atgctatttc cgcaagcaag cgtggaatta aaacaattat cgctggagct
     2161 ggtggggctg ctcacttgcc aggtatggtg gctgcaatga caccacttcc tgtcatcggt
     2221 gtgcccgtaa aaggttcttg tctagatgga gtagattctt tacattcaat tgtgcaaatg
     2281 cctagaggtg ttccagtagc taccgtcgct attaataata gtacgaacgc tgcgctgttg
     2341 gctgtcagac tgcttggcgc ttatgattca agttatacaa cgaaaatgga acagttttta
     2401 ttaaagcaag aagaagaagt tcttgtcaaa gcacaaaagt tagaaactgt cggttacgaa
     2461 gcttatctag aaaacaagta atatataagt ttattgatat acttgtacag caaataatta
     2521 taaaatgata tacctatttt ttaggctttg ttatgattac atcaaatgtg gacttcatac
     2581 atagaaatca acgcttacag gtgtcctttt ttaagaattt catacataag atctatgcgg
     2641 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta
     2701 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2761 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2821 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2881 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2941 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3001 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3061 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3121 atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3181 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3241 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3301 gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc tcgaggtcga
     3361 cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta gttctagagc
     3421 ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta attgcgcgct
     3481 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3541 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3601 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3661 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3721 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3781 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3841 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3901 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3961 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4021 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4081 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4141 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4201 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4261 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4321 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4381 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4441 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4501 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4561 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4621 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4681 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4741 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4801 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4861 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4921 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4981 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5041 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5101 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5161 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5221 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5281 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5341 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5401 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5461 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5521 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5581 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5641 cacctgacgt ctaacacagt cctttcccgc aattttcttt ttctattact cttggcctcc
     5701 tctagtacac tctatatttt tttatgcctc ggtaatgatt ttcatttttt tttttccacc
     5761 tagcggatga ctcttttttt ttcttagcga ttggcattat cacataatga attatacatt
     5821 atataaagta atgtgatttc ttcgaagaat atactaaaaa atgagcaggc aagataaacg
     5881 aaggcaaaga tgacagagca gaaagcccta gtaaagcgta ttacaaatga aaccaagatt
     5941 cagattgcga tctctttaaa gggtggtccc ctagcgatag agcactcgat cttcccagaa
     6001 aaagaggcag aagcagtagc agaacaggcc acacaatcgc aagtgattaa cgtccacaca
     6061 ggtatagggt ttctggacca tatgatacat gctctggcca agcattccgg ctggtcgcta
     6121 atcgttgagt gcattggtga cttacacata gacgaccatc acaccactga agactgcggg
     6181 attgctctcg gtcaagcttt taaagaggcc ctaggggccg tgcgtggagt aaaaaggttt
     6241 ggatcaggat ttgcgccttt ggatgaggca ctttccagag cggtggtaga tctttcgaac
     6301 aggccgtacg cagttgtcga acttggtttg caaagggaga aagtaggaga tctctcttgc
     6361 gagatgatcc cgcattttct tgaaagcttt gcagaggcta gcagaattac cctccacgtt
     6421 gattgtctgc gaggcaagaa tgatcatcac cgtagtgaga gtgcgttcaa ggctcttgcg
     6481 gttgccataa gagaagccac ctcgcccaat ggtaccaacg atgttccctc caccaaaggt
     6541 gttcttatgt agtgacaccg attatttaaa gctgcagcat acgatatata tacatgtgta
     6601 tatatgtata cctatgaatg tcagtaagta tgtatacgaa cagtatgata ctgaagatga
     6661 caaggtaatg catcattcta tacgtgtcat tctgaacgag gcatcatgac attaacctat
     6721 aa
//
