LOCUS       sequence-239184-        5906 bp    DNA     circular SYN 14-APR-2025
DEFINITION  TRP1-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 5906)
  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 5906)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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 TRP1-HphMx
FEATURES             Location/Qualifiers
     source          1..5906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(7..29)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     129..148
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        169..450
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             451..1125
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1222..1677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1359..1380)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1571..1590
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1654..2034)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1803..1825
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1817..1834
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1818..1834
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1844..1863
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1844..1862
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1871..1978
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1887..1903
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1888..1904
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1938..1954)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1938..1954)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1991..2011)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1991..2009)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2030..2046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2030..2046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2043..2065)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2054..2070
                     /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(2078..2084,2085..2102,2103..2108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2123..2144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2261..2278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2432..3020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2512..2531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3191..3982,3983..4051))
                     /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     3814..3833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4052..4156)
                     /gene="bla"
                     /label=AmpR promoter
     gene            4229..5804
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        4229..4572
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             4573..5601
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             4573..5601
                     /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      5607..5804
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     5857..5875
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
       61 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
      121 taactatgcg gcatcagagc agattgtact gagagtgcac catagatcaa cgacattact
      181 atatatataa tataggaagc atttaataga acagcatcgt aatatatgtg tactttgcag
      241 ttatgacgcc agatggcagt agtggaagat attctttatt gaaaaatagc ttgtcacctt
      301 acgtacaatc ttgatccgga gcttttcttt ttttgccgat taagaattaa ttcggtcgaa
      361 aaaagaaaag gagagggcca agagggaggg cattggtgac tattgagcac gtgagtatac
      421 gtgattaagc acacaaaggc agcttggagt atgtctgtta ttaatttcac aggtagttct
      481 ggtccattgg tgaaagtttg cggcttgcag agcacagagg ccgcagaatg tgctctagat
      541 tccgatgctg acttgctggg tattatatgt gtgcccaata gaaagagaac aattgacccg
      601 gttattgcaa ggaaaatttc aagtcttgta aaagcatata aaaatagttc aggcactccg
      661 aaatacttgg ttggcgtgtt tcgtaatcaa cctaaggagg atgttttggc tctggtcaat
      721 gattacggca ttgatatcgt ccaactgcat ggagatgagt cgtggcaaga ataccaagag
      781 ttcctcggtt tgccagttat taaaagactc gtatttccaa aagactgcaa catactactc
      841 agtgcagctt cacagaaacc tcattcgttt attcccttgt ttgattcaga agcaggtggg
      901 acaggtgaac ttttggattg gaactcgatt tctgactggg ttggaaggca agagagcccc
      961 gaaagcttac attttatgtt agctggtgga ctgacgccag aaaatgttgg tgatgcgctt
     1021 agattaaatg gcgttattgg tgttgatgta agcggaggtg tggagacaaa tggtgtaaaa
     1081 gactctaaca aaatagcaaa tttcgtcaaa aatgctaaga aataggttat tactgagtag
     1141 tatttattta attatatcag ttattaccct atgcggtgtg aaataccgca cagatgcgta
     1201 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1261 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1321 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1381 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1441 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1501 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1561 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1621 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     1681 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     1741 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     1801 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     1861 gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga
     1921 attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     1981 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     2041 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     2101 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     2161 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2221 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2281 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2341 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2401 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2461 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2521 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2581 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2641 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2701 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2761 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2821 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2881 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2941 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3001 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3061 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3121 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3181 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3241 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3301 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3361 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3421 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3481 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3541 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3601 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3661 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3721 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3781 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3841 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3901 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3961 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4021 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4081 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4141 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcttg tttagcttgc
     4201 ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag
     4261 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
     4321 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
     4381 ttacggctcc tcgctgcgga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
     4441 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
     4501 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
     4561 acataaacaa ccatgggtaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     4621 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     4681 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     4741 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     4801 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     4861 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     4921 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     4981 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     5041 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     5101 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     5161 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     5221 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     5281 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     5341 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     5401 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     5461 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     5521 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     5581 cgtccgaggg caaaggaata atcagtactg acaataaaaa gattcttgtt ttcaagaact
     5641 tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga
     5701 tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag
     5761 taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact
     5821 aacgccgcca tccagatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     5881 cgtctcgcgc gtttcggtga tgacgg
//
