LOCUS       sequence-239178-        7319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2-HphMx 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 7319)
  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 7319)
  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-HphMx
FEATURES             Location/Qualifiers
     source          1..7319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..49
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        240..615
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             616..2331
                     /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(2536..2991)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2673..2694)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2885..2904
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2968..3348)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3117..3139
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3131..3148
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3132..3148
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3158..3177
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3158..3176
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3185..3292
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3201..3217
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3202..3218
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3252..3268)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3252..3268)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3305..3325)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3305..3323)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3344..3360)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3344..3360)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3357..3379)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3368..3384
                     /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(3392..3398,3399..3416,3417..3422))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3437..3458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3575..3592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3746..4334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3826..3845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4505..5296,5297..5365))
                     /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     5128..5147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5366..5470)
                     /gene="bla"
                     /label=AmpR promoter
     gene            5543..7118
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        5543..5886
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             5887..6915
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             5887..6915
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      6921..7118
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     7171..7189
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7227..7249)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
       61 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
      121 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
      181 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagat ctgaattaat
      241 tcttgaataa tacataactt ttcttaaaag aatcaaagac agataaaatt taagagatat
      301 taaatattag tgagaagccg agaattttgt aacaccaaca taacactgac atctttaaca
      361 acttttaatt atgatacatt tcttacgtca tgattgatta ttacagctat gctgacaaat
      421 gactcttgtt gcatggctac gaaccgggta atactaagtg attgactctt gctgaccttt
      481 tattaagaac taaatggaca atattatgga gcatttcatg tataaattgg tgcgtaaaat
      541 cgttggatct ctcttctaag tacatcctac tataacaatc aagaaaaaca agaaaatcgg
      601 acaaaacaat caagtatgga ttctagaaca gttggtatat taggaggggg acaattggga
      661 cgtatgattg ttgaggcagc aaacaggctc aacattaaga cggtaatact agatgctgaa
      721 aattctcctg ccaaacaaat aagcaactcc aatgaccacg ttaatggctc cttttccaat
      781 cctcttgata tcgaaaaact agctgaaaaa tgtgatgtgc taacgattga gattgagcat
      841 gttgatgttc ctacactaaa gaatcttcaa gtaaaacatc ccaaattaaa aatttaccct
      901 tctccagaaa caatcagatt gatacaagac aaatatattc aaaaagagca tttaatcaaa
      961 aatggtatag cagttaccca aagtgttcct gtggaacaag ccagtgagac gtccctattg
     1021 aatgttggaa gagatttggg ttttccattc gtcttgaagt cgaggacttt ggcatacgat
     1081 ggaagaggta acttcgttgt aaagaataag gaaatgattc cggaagcttt ggaagtactg
     1141 aaggatcgtc ctttgtacgc cgaaaaatgg gcaccattta ctaaagaatt agcagtcatg
     1201 attgtgaggt ctgttaacgg tttagtgttt tcttacccaa ttgtagagac tatccacaag
     1261 gacaatattt gtgacttatg ttatgcgcct gctagagttc cggactccgt tcaacttaag
     1321 gcgaagttgt tggcagaaaa tgcaatcaaa tcttttcccg gttgtggtat atttggtgtg
     1381 gaaatgttct atttagaaac aggggaattg cttattaacg aaattgcccc aaggcctcac
     1441 aactctggac attataccat tgatgcttgc gtcacttctc aatttgaagc tcatttgaga
     1501 tcaatattgg atttgccaat gccaaagaat ttcacatctt tctccaccat tacaacgaac
     1561 gccattatgc taaatgttct tggagacaaa catacaaaag ataaagagct agaaacttgc
     1621 gaaagagcat tggcgactcc aggttcctca gtgtacttat atggaaaaga gtctagacct
     1681 aacagaaaag taggtcacat aaatattatt gcctccagta tggcggaatg tgaacaaagg
     1741 ctgaactaca ttacaggtag aactgatatt ccaatcaaaa tctctgtcgc tcaaaagttg
     1801 gacttggaag caatggtcaa accattggtt ggaatcatca tgggatcaga ctctgacttg
     1861 ccggtaatgt ctgccgcatg tgcggtttta aaagattttg gcgttccatt tgaagtgaca
     1921 atagtctctg ctcatagaac tccacatagg atgtcagcat atgctatttc cgcaagcaag
     1981 cgtggaatta aaacaattat cgctggagct ggtggggctg ctcacttgcc aggtatggtg
     2041 gctgcaatga caccacttcc tgtcatcggt gtgcccgtaa aaggttcttg tctagatgga
     2101 gtagattctt tacattcaat tgtgcaaatg cctagaggtg ttccagtagc taccgtcgct
     2161 attaataata gtacgaacgc tgcgctgttg gctgtcagac tgcttggcgc ttatgattca
     2221 agttatacaa cgaaaatgga acagttttta ttaaagcaag aagaagaagt tcttgtcaaa
     2281 gcacaaaagt tagaaactgt cggttacgaa gcttatctag aaaacaagta atatataagt
     2341 ttattgatat acttgtacag caaataatta taaaatgata tacctatttt ttaggctttg
     2401 ttatgattac atcaaatgtg gacttcatac atagaaatca acgcttacag gtgtcctttt
     2461 ttaagaattt catacataag atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     2521 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2581 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2641 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2701 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2761 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2821 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2881 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2941 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc
     3001 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     3061 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     3121 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     3181 attgggtacc gggccccccc tcgaggtcga cggtatcgat aagcttgata tcgaattcct
     3241 gcagcccggg ggatccacta gttctagagc ggccgccacc gcggtggagc tccagctttt
     3301 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     3361 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3421 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3481 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3541 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3601 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3661 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3721 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3781 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3841 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3901 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3961 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4021 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4081 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4141 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4201 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4261 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4321 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4381 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4441 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4501 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4561 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4621 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4681 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4741 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4801 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4861 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4921 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4981 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5041 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5101 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5161 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5221 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5281 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5341 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5401 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5461 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cttgtttagc ttgcctcgtc
     5521 cccgccgggt cacccggcca gcgacatgga ggcccagaat accctccttg acagtcttga
     5581 cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac atttagccca tacatcccca
     5641 tgtataatca tttgcatcca tacattttga tggccgcacg gcgcgaagca aaaattacgg
     5701 ctcctcgctg cggacctgcg agcagggaaa cgctcccctc acagacgcgt tgaattgtcc
     5761 ccacgccgcg cccctgtaga gaaatataaa aggttaggat ttgccactga ggttcttctt
     5821 tcatatactt ccttttaaaa tcttgctagg atacagttct cacatcacat ccgaacataa
     5881 acaaccatgg gtaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     5941 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     6001 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     6061 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     6121 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     6181 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     6241 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     6301 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     6361 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     6421 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     6481 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     6541 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     6601 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     6661 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     6721 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     6781 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     6841 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     6901 agggcaaagg aataatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     6961 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     7021 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     7081 catgcgtcaa tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc
     7141 gccatccaga tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     7201 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     7261 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgt
//
