LOCUS       sequence-239189-        5450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TRP1-NatMx 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 5450)
  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 5450)
  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 TRP1-NatMx
FEATURES             Location/Qualifiers
     source          1..5450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      397..594
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     647..665
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(703..725)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     825..844
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        865..1146
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1147..1821
                     /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(1918..2373)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2055..2076)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2267..2286
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2350..2730)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2499..2521
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2513..2530
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2514..2530
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2540..2559
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2540..2558
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2567..2674
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2583..2599
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2584..2600
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2634..2650)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2634..2650)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2687..2707)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2687..2705)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2726..2742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2726..2742)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2739..2761)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2750..2766
                     /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(2774..2780,2781..2798,2799..2804))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2819..2840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2957..2974)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3128..3716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3208..3227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3887..4678,4679..4747))
                     /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     4510..4529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4748..4852)
                     /gene="bla"
                     /label=AmpR promoter
     gene            join(4925..5450,1..594)
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        4925..5268
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             join(5269..5450,1..391)
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
ORIGIN
        1 cgacgacgaa tcggacgacg aatcggacga cggggaggac ggcgacccgg actcccggac
       61 gttcgtcgcg tacggggacg acggcgacct ggcgggcttc gtggtcgtct cgtactccgg
      121 ctggaaccgc cggctgaccg tcgaggacat cgaggtcgcc ccggagcacc gggggcacgg
      181 ggtcgggcgc gcgttgatgg ggctcgcgac ggagttcgcc cgcgagcggg gcgccgggca
      241 cctctggctg gaggtcacca acgtcaacgc accggcgatc cacgcgtacc ggcggatggg
      301 gttcaccctc tgcggcctgg acaccgccct gtacgacggc accgcctcgg acggcgagca
      361 ggcgctctac atgagcatgc cctgccccta atcagtactg acaataaaaa gattcttgtt
      421 ttcaagaact tgtcatttgt atagtttttt tatattgtag ttgttctatt ttaatcaaat
      481 gttagcgtga tttatatttt ttttcgcctc gacatcatct gcccagatgc gaagttaagt
      541 gcgcagaaag taatatcatg cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg
      601 attcgatact aacgccgcca ttattatcat gacattaacc tataaaaata ggcgtatcac
      661 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
      721 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
      781 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
      841 tgtactgaga gtgcaccata gatcaacgac attactatat atataatata ggaagcattt
      901 aatagaacag catcgtaata tatgtgtact ttgcagttat gacgccagat ggcagtagtg
      961 gaagatattc tttattgaaa aatagcttgt caccttacgt acaatcttga tccggagctt
     1021 ttcttttttt gccgattaag aattaattcg gtcgaaaaaa gaaaaggaga gggccaagag
     1081 ggagggcatt ggtgactatt gagcacgtga gtatacgtga ttaagcacac aaaggcagct
     1141 tggagtatgt ctgttattaa tttcacaggt agttctggtc cattggtgaa agtttgcggc
     1201 ttgcagagca cagaggccgc agaatgtgct ctagattccg atgctgactt gctgggtatt
     1261 atatgtgtgc ccaatagaaa gagaacaatt gacccggtta ttgcaaggaa aatttcaagt
     1321 cttgtaaaag catataaaaa tagttcaggc actccgaaat acttggttgg cgtgtttcgt
     1381 aatcaaccta aggaggatgt tttggctctg gtcaatgatt acggcattga tatcgtccaa
     1441 ctgcatggag atgagtcgtg gcaagaatac caagagttcc tcggtttgcc agttattaaa
     1501 agactcgtat ttccaaaaga ctgcaacata ctactcagtg cagcttcaca gaaacctcat
     1561 tcgtttattc ccttgtttga ttcagaagca ggtgggacag gtgaactttt ggattggaac
     1621 tcgatttctg actgggttgg aaggcaagag agccccgaaa gcttacattt tatgttagct
     1681 ggtggactga cgccagaaaa tgttggtgat gcgcttagat taaatggcgt tattggtgtt
     1741 gatgtaagcg gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat agcaaatttc
     1801 gtcaaaaatg ctaagaaata ggttattact gagtagtatt tatttaatta tatcagttat
     1861 taccctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa
     1921 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1981 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2041 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2101 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2161 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2221 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     2281 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     2341 ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg
     2401 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2461 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2521 gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta ccgggccccc
     2581 cctcgaggtc gacggtatcg ataagcttga tatcgaattc ctgcagcccg ggggatccac
     2641 tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
     2701 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2761 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2821 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2881 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2941 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3001 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3061 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3121 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3181 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3241 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3301 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3361 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3421 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3481 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3541 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3601 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3661 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3721 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3781 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3841 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3901 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3961 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4021 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4081 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4141 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4201 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4261 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4321 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4381 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4441 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4501 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4561 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4621 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4681 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4741 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4801 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4861 cccgaaaagt gccacctgac gtcttgttta gcttgcctcg tccccgccgg gtcacccggc
     4921 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
     4981 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
     5041 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
     5101 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
     5161 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
     5221 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaccact
     5281 cttgacgaca cggcttaccg gtaccgcacc agtgtcccgg gggacgccga ggccatcgag
     5341 gcactggatg ggtccttcac caccgacacc gtcttccgcg tcaccgccac cggggacggc
     5401 ttcaccctgc gggaggtgcc ggtggacccg cccctgacca aggtgttccc
//
