LOCUS       sequence-360111-        6856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  YIp for expressing Murine polyomavirus deltaVP1 (GAL1 promoter) and 
            mRuby3 tagged with VP2C (GAL10 promoter). mRuby3 was codon-optimised
            for E. coli. Contains uracil marker (K. lactis URA3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6856)
  AUTHORS   Cheah LC, Liu L, Plan MR, Peng B, Lu Z, Schenk G, Vickers CE, 
            Sainsbury F
  TITLE     Synthetic in vivo compartmentalisation improves metabolic flux and 
            modulates the product profile of promiscuous enzymes
  JOURNAL   bioRxiv 2022.11.24.517869
REFERENCE   2  (bases 1 to 6856)
  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: "bioRxiv 
            2022.11.24.517869"
COMMENT     Sequence Label: mRuby3 only VLP
FEATURES             Location/Qualifiers
     source          1..6856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(246..262)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(246..262)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(259..281)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    270..286
                     /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(294..300,301..318,319..324))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    339..360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(477..494)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(648..1236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(728..747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1407..2198,2199..2267))
                     /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     2030..2049
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2268..2372)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2440..2458
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2496..2518)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2618..2637
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2831..2853
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2845..2862
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2846..2862
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4120..4830)
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     promoter        5254..5695
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    5254..5371
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5666..5689
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 aagcggccgc actagtagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
       61 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
      121 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
      181 atcagttatt acccgggaat ctcggtcgta atgatttaaa tgcatgcaag cttggcgtaa
      241 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
      301 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      361 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
      421 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
      481 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
      541 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      601 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
      661 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      721 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      781 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      841 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      901 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      961 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1021 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1081 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1141 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     1201 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     1261 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     1321 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     1381 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     1441 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1501 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1561 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1621 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1681 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1741 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1801 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1861 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1921 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1981 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2041 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2101 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2161 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     2221 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     2281 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     2341 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     2401 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     2461 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     2521 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     2581 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     2641 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     2701 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     2761 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     2821 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcat ttaaattttt
     2881 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     2941 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     3001 tgcacagaac aaaaacctgc aggaaacgaa gataaaacat gtccacaaaa tcatatacca
     3061 gtagagctga gactcatgca agtccggttg catcgaaact tttacgttta atggatgaaa
     3121 agaagaccaa tttgtgtgct tctcttgacg ttcgttcgac tgatgagcta ttgaaacttg
     3181 ttgaaacgtt gggtccatac atttgccttt tgaaaacaca cgttgatatc ttggatgatt
     3241 tcagttatga gggtactgtc gttccattga aagcattggc agagaaatac aagttcttga
     3301 tatttgagga cagaaaattc gccgatatcg gtaacacagt caaattacaa tatacatcgg
     3361 gcgtttaccg tatcgcagaa tggtctgata tcaccaacgc ccacggggtt actggtgctg
     3421 gtattgttgc tggcttgaaa caaggtgcgc aagaggtcac caaagaacca aggggattat
     3481 tgatgcttgc tgaattgtct tccaagggtt ctctagcaca cggtgaatat actaagggta
     3541 ccgttgatat tgcaaagagt gataaagatt tcgttattgg gttcattgct cagaacgata
     3601 tgggaggaag agaagaaggg tttgattggc taatcatgac cccaggtgta ggtttagacg
     3661 acaaaggcga tgcattgggt cagcagtaca gaaccgtcga cgaagttgta agtggtggat
     3721 cagatatcat cattgttggc agaggacttt tcgccaaggg tagagatcct aaggttgaag
     3781 gtgaaagata cagaaatgct ggatgggaag cgtaccaaaa gagaatcagc gctccccatt
     3841 aattatacag gaaacttaat agaacaaatc acatatttaa tctaatagcc acctgcattg
     3901 gcacggtgca acactacttc aacttcatct tacaaaaaga tcacgtgatc tgttgtattg
     3961 ggatctctag agtaacaata tcatgagacc ttttatagaa gtggcgccaa aactaaatgt
     4021 atttgaaaat acaaaaaacg cacaggtttc tttatatatg tatatatata tatatatata
     4081 tataaaactg tacatatact gtttaaatta atctatttac ttgtacagct cgtccatgcc
     4141 accaccaagg ttgctgtatt tggcaactgc cacttcgcgt tgcactacgt aggtttcatt
     4201 gtcactctcc tcgatccttt ccaggcggtg atcaacggca tggacaccgg gcatcttgat
     4261 gttcccgacg gtcttttttg acctgtaagt tgtcacgaag ttgcagtgca gatggccacc
     4321 accatcaact ttcagtgcga tgtcagtgta tcctctcaga ccaccatctg ctggatacat
     4381 catctctgta ttaggctccc aacccttggt cttcttctgc atcacgggac cattggaggg
     4441 aaagtttacc cctctgacct tgacgttgta gacgagctcg ccatcctcaa ggctggtgtc
     4501 ctgcgtgacg gtgacgactc caccatcttc gtatctcgta actctttccc aagtaaaacc
     4561 ctcaggaaag gactgtttaa agaaatcagg gatgtcggcc gggtacttga taaaagtacg
     4621 gctgccatac atgaacgacg tggcaagaat gtcaaaggca aatggcaggg gtcctccctc
     4681 gatgactttg atcctcatgg tttgcactcc ctcgtacggt ctgccttctc cttcacctgt
     4741 gcatttgaat tggtggccgt tgaccgaacc ttccatgacc accttcatac gcatattttc
     4801 cttgatcagc tcttcgccct tagacaccat agaaccgccg ccaccggagc caccgccacc
     4861 agaaccacca ccacctggag tgatgtcacc atacaaaccc aaaatcaatg gcaacatcca
     4921 atctggagta actctttgat gagaaacaac ttgagcttga taaaatttaa taacttcacc
     4981 agattcatct tgaatatgag cagctggagt tggaccacca ttacccattt tatcgattta
     5041 tattgaattt tcaaaaattc ttactttttt ttttggatgg acgcaaagaa gtttaataat
     5101 catattacat ggcaatacca ccatatacat atccatatct aatcttactt atatgttgtg
     5161 gaaatgtaaa gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta
     5221 atacgcttaa ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggcgac
     5281 agccctccga cggaagactc tcctccgtgc gtcctggtct tcaccggtcg cgttcctgaa
     5341 acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt
     5401 tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc aaatcaacga
     5461 atcaaattaa caaccatagg ataataatgc gattagtttt ttagccttat ttctggggta
     5521 attaatcagc gaagcgatga tttttgatct attaacagat atataaatgc aaaagctgca
     5581 taaccacttt aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgtc
     5641 ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa
     5701 actatagcta gcaaaatggc ttcaggtgta agtaaatgcg aaaccaaatg cacaaaggct
     5761 tgccctagac cagctccagt tcctaagtta ttgattaaag gtggtatgga agttttggat
     5821 ttggttacag gtccagattc tgttactgaa atcgaagcat ttttgaaccc aagaatgggt
     5881 caaccaccaa caccagaatc tttaactgaa ggtggtcaat attacggttg gtcaagaggt
     5941 attaatttgg ctacatctga tactgaagat tcaccaggta ataatacatt accaacttgg
     6001 tctatggcaa aattgcaatt accaatgttg aacgaagatt tgacttgtga tactttgcaa
     6061 atgtgggaag cagtttcagt taaaacagaa gttgttggtt ctggttcttt gttggatgtt
     6121 catggtttta ataagccaac agatactgtt aacacaaagg gtatttctac tccagttgaa
     6181 ggttcacaat atcatgtttt tgctgttggt ggtgaaccat tggatttgca aggtttagtt
     6241 acagatgcaa gaactaagta caaggaagaa ggtgttgtta ctattaaaac aatcactaag
     6301 aaagatatgg ttaataagga tcaagttttg aacccaattt caaaggctaa attggataag
     6361 gatggcatgt acccagttga aatttggcat ccagatccag ctaaaaatga aaacacaaga
     6421 tacttcggta attacactgg tggtactaca actccaccag ttttgcaatt cactaacact
     6481 ttgacaactg ttttgttaga tgaaaatggt gttggtccat tatgtaaagg tgaaggtttg
     6541 tacttatctt gcgttgatat tatgggttgg agagttacaa gaaactacga tgttcatcat
     6601 tggagaggtt tgccaagata cttcaagatt actttaagaa aaagatgggt taaaaatcca
     6661 tacccaatgg cttctttgat ttcttctttg tttaataata tgttaccaca agttcaaggt
     6721 caaccaatgg aaggtgaaaa cacacaagtt gaagaagtta gagtttacga tggtactgaa
     6781 ccagttccag gtgaccctga catgacaaga tatgtagata gattcggtaa aactaaaact
     6841 gtattccctg gtaatt
//
