LOCUS       sequence-360119-        9877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  YIp for expressing Murine polyomavirus deltaVP1 (GAL1 promoter), 
            VP2C-ERG20(F96W-N127W) (GAL10 promoter), and VP2C-AcNES1 (GAL7 
            promoter). Contains uracil marker (K. lactis URA3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9877)
  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 9877)
  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: Coexpressed VLP NES (M)
FEATURES             Location/Qualifiers
     source          1..9877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1301..1742)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(1307..1330)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1625..1742
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(7155..7172)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7155..7171)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7164..7186)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7380..7399)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7499..7521
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7559..7577)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7645..7749
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7750..7818,7819..8610)
                     /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     complement(7968..7987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8781..9369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9270..9289
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9523..9540
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9657..9678
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9693..9698,9699..9716,9717..9723)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9731..9747
                     /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)."
     primer_bind     9736..9758
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9755..9771
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9755..9771
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc
       61 cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt
      121 cctactagtg cggccgctta attaccaggg aatacagttt tagttttacc gaatctatct
      181 acatatcttg tcatgtcagg gtcacctgga actggttcag taccatcgta aactctaact
      241 tcttcaactt gtgtgttttc accttccatt ggttgacctt gaacttgtgg taacatatta
      301 ttaaacaaag aagaaatcaa agaagccatt gggtatggat ttttaaccca tctttttctt
      361 aaagtaatct tgaagtatct tggcaaacct ctccaatgat gaacatcgta gtttcttgta
      421 actctccaac ccataatatc aacgcaagat aagtacaaac cttcaccttt acataatgga
      481 ccaacaccat tttcatctaa caaaacagtt gtcaaagtgt tagtgaattg caaaactggt
      541 ggagttgtag taccaccagt gtaattaccg aagtatcttg tgttttcatt tttagctgga
      601 tctggatgcc aaatttcaac tgggtacatg ccatccttat ccaatttagc ctttgaaatt
      661 gggttcaaaa cttgatcctt attaaccata tctttcttag tgattgtttt aatagtaaca
      721 acaccttctt ccttgtactt agttcttgca tctgtaacta aaccttgcaa atccaatggt
      781 tcaccaccaa cagcaaaaac atgatattgt gaaccttcaa ctggagtaga aatacccttt
      841 gtgttaacag tatctgttgg cttattaaaa ccatgaacat ccaacaaaga accagaacca
      901 acaacttctg ttttaactga aactgcttcc cacatttgca aagtatcaca agtcaaatct
      961 tcgttcaaca ttggtaattg caattttgcc atagaccaag ttggtaatgt attattacct
     1021 ggtgaatctt cagtatcaga tgtagccaaa ttaatacctc ttgaccaacc gtaatattga
     1081 ccaccttcag ttaaagattc tggtgttggt ggttgaccca ttcttgggtt caaaaatgct
     1141 tcgatttcag taacagaatc tggacctgta accaaatcca aaacttccat accaccttta
     1201 atcaataact taggaactgg agctggtcta gggcaagcct ttgtgcattt ggtttcgcat
     1261 ttacttacac ctgaagccat tttgctagct atagtttttt ctccttgacg ttaaagtata
     1321 gaggtatatt aacaattttt tgttgatact tttatgacat ttgaataaga agtaatacaa
     1381 accgaaaatg ttgaaagtat tagttaaagt ggttatgcag cttttgcatt tatatatctg
     1441 ttaatagatc aaaaatcatc gcttcgctga ttaattaccc cagaaataag gctaaaaaac
     1501 taatcgcatt attatcctat ggttgttaat ttgattcgtt gatttgaagg tttgtggggc
     1561 caggttactg ccaatttttc ctcttcataa ccataaaagc tagtattgta gaatctttat
     1621 tgttcggagc agtgcggcgc gaggcacatc tgcgtttcag gaacgcgacc ggtgaagacc
     1681 aggacgcacg gaggagagtc ttccgtcgga gggctgtcgc ccgctcggcg gcttctaatc
     1741 cgtacttcaa tatagcaatg agcagttaag cgtattactg aaagttccaa agagaaggtt
     1801 tttttaggct aagataatgg ggctctttac atttccacaa catataagta agattagata
     1861 tggatatgta tatggtggta ttgccatgta atatgattat taaacttctt tgcgtccatc
     1921 caaaaaaaaa agtaagaatt tttgaaaatt caatataaat cgataaaatg ggtaatggtg
     1981 gtccaactcc agctgctcat attcaagatg aatctggtga agttattaaa ttttatcaag
     2041 ctcaagttgt ttctcatcaa agagttactc cagattggat gttgccattg attttgggtt
     2101 tgtatggtga catcactcca ggtggtggtg gttctggtgg cggtggctcc ggtggcggcg
     2161 gttctggatc catggcttca gaaaaagaaa ttaggagaga gagattcttg aacgttttcc
     2221 ctaaattagt agaggaattg aacgcatcgc ttttggctta cggtatgcct aaggaagcat
     2281 gtgactggta tgcccactca ttgaactaca acactccagg cggtaagcta aatagaggtt
     2341 tgtccgttgt ggacacgtat gctattctct ccaacaagac cgttgaacaa ttggggcaag
     2401 aagaatacga aaaggttgcc attctaggtt ggtgcattga gttgttgcag gcttactggt
     2461 tggtcgccga tgatatgatg gacaagtcca ttaccagaag aggccaacca tgttggtaca
     2521 aggttcctga agttggggaa attgccatct gggacgcatt catgttagag gctgctatct
     2581 acaagctttt gaaatctcac ttcagaaacg aaaaatacta catagatatc accgaattgt
     2641 tccatgaggt caccttccaa accgaattgg gccaattgat ggacttaatc actgcacctg
     2701 aagacaaagt cgacttgagt aagttctccc taaagaagca ctccttcata gttactttca
     2761 agactgctta ctattctttc tacttgcctg tcgcattggc catgtacgtt gccggtatca
     2821 cggatgaaaa ggatttgaaa caagccagag atgtcttgat tccattgggt gaatacttcc
     2881 aaattcaaga tgactactta gactgcttcg gtaccccaga acagatcggt aagatcggta
     2941 cagatatcca agataacaaa tgttcttggg taatcaacaa ggcattagaa cttgcttccg
     3001 cagaacaaag aaagacttta gacgaaaatt acggtaagaa ggactcagtc gcagaagcca
     3061 aatgcaaaaa gattttcaat gacttgaaaa ttgaacagct ataccacgaa tatgaagagt
     3121 ctattgccaa ggatttgaag gccaaaattt ctcaggtcga tgagtctcgt ggcttcaaag
     3181 ctgatgtctt aactgcgttc ttgaacaaag tttacaagag aagcaaataa agatcttttg
     3241 ccagcttact atccttcttg aaaatatgca ctctatatct tttagttctt aattgcaaca
     3301 catagatttg ctgtataacg aattttatgc tattttttaa atttggagtt cagtgataaa
     3361 agtgtcacag cgaatttcct cacatgtagg gaccgaattg tttacaagtt ctctgtacca
     3421 ccatggagac atcaaaaatt gaaaatctat ggaaagatat ggacggtagc aacaagaata
     3481 tagcacgagc cgcggagttc atttcgttac ttttgatatc actcacaact attgcgaagc
     3541 gcttcagtga aaaaatcata aggaaaagtt gtaaatatta ttggtagtat tcgtttggta
     3601 aagtagaggg ggtaattttt cccctttatt ttgttcatac attcttaaat tgctttgcct
     3661 ctccttttgg aaagctatac ttcggagcac tgttgagcga aggctcatta gatatatttt
     3721 ctgtcatttt ccttaaccca aaaataaggg aaagggtcca aaaagcgctc ggacaactgt
     3781 tgaccgtgat ccgaaggact ggctatacag tgttcacaaa atagccaagc tgaaaataat
     3841 gtgtagctat gttcagttag tttggctagc aaagatataa aagcaggtcg gaaatattta
     3901 tgggcattat tatgcagagc atcaacatga taaaaaaaaa cagttgaata ttccctcaaa
     3961 aatcgataaa atgggtaatg gtggtccaac tccagctgct catattcaag atgaatctgg
     4021 tgaagttatt aaattttatc aagctcaagt tgtttctcat caaagagtta ctccagattg
     4081 gatgttgcca ttgattttgg gtttgtatgg tgacatcact ccaggtggtg gtggttctgg
     4141 tggcggtggc tccggtggcg gcggttctga gctcatggct accgcagcag gtcctatcgc
     4201 aactaacaac tccccacaaa actccaacgc ttacagaact ccaatcgctc cttccgtacc
     4261 aattactcat aaatggtcta tagctgaaga tttgacatgt atttccaatc ctagtaagca
     4321 caataaccct caaactggtt acagatcatt ttctgacgaa ttatacgtta agtacgaaga
     4381 aaagttggaa gatgttagaa aagcattaag agaagttgaa gaaaaccctt tggaaggttt
     4441 agttatgata gacgctttgc aaagattggg tatcgattac catttcagag gtgaaattgg
     4501 tgcattcttg caaaagcaac aaatcatatc ttcaactcca gatggttacc ctgaacatgg
     4561 tttgtacgaa gtttcaacat tgtttagatt cttaagacaa gaaggtcaca atgttaccgc
     4621 tgacgtcttt aataacttca aggataagga aggtagattc agatcagaat tgtcaacaga
     4681 tattagaggt ttgatgtcct tatacgaagc aagtcaattg agaatagaag gtgaagacat
     4741 cttagatcaa gctgctgatt tctccagtca attgttaggt agatggacaa aagatcctaa
     4801 tcatcacgaa gccagattgg tttctaacac tttaacacat ccataccaca agtcattggc
     4861 tacctttatg ggtcaaaaat tgtcctacat gaactgcaag ggtccaaact gggacggtgt
     4921 cgataatttg caagaattag ctaagatgga tttgactatc gtacaaagta tccatcaaaa
     4981 ggaagtattc caagtttctc aatggtggaa ggatacaggt ttagccaatg aattgaaatt
     5041 ggctagaaac caaccattga agtggtatat gtggcctatg gccgctttaa ccgatccaag
     5101 attcagtgaa gaaagagttg aattaactaa gcctatttct tttatataca tcatagatga
     5161 catcttcgac gtctatggta ccattgaaga attgaccttg tttactgatg cagttaatag
     5221 atgggaattg tctgccgtcg aacaattacc agactacatg aaagtatgtt tcaaggcttt
     5281 gtacgatgtt accaacgaaa tcgcatacaa aatctataaa aagcatggtc aaaaccctat
     5341 tgattccttg caaaagactt gggctagttt gtgcaatgca tttttagttg aagcaaagtg
     5401 gttcgcctct ggtcacttgc caaatgcaga agaatactta aagaacggta tcatctcttc
     5461 aggtgttcat gttgtcttgg cccacatgtt tttcttgtta ggtgacggta ttacacaaga
     5521 atcagttgat ttggtagatg actatccagg tatttccaca agtatcgcaa ccattttgag
     5581 attatctgat gacttgggtt cagccaaaga tgaagaccaa gatggttatg atggttctta
     5641 catcgaatgt tacatgaagg aacataaggg ttccagtgtc gattcagcca gagaagaagt
     5701 aataagaatg atctccgaag catggaaatg tttgaataag gaatgcttat caccaaaccc
     5761 tttttctgaa tcattcagaa taggttcctt gaatatggct agaatgatcc ctatgatgta
     5821 ctcttacgat gacaaccata acttgccaat tttagaagaa cacatgaagg caatgatcta
     5881 taacacatct ttataactcg agatagatta atttaaacag tatatgtaca gttttatata
     5941 tatatatata tatatacata tataaagaaa cctgtgcgtt ttttgtattt tcaaatacat
     6001 ttagttttgg cgccacttct ataaaaggtc tcatgatatt gttactctag agatcccaat
     6061 acaacagatc acgtgatctt tttgtaagat gaagttgaag tagtgttgca ccgtgccaat
     6121 gcaggtggct attagattaa atatgtgatt tgttctatta agtttcctgt ataattaatg
     6181 gggagcgctg attctctttt ggtacgcttc ccatccagca tttctgtatc tttcaccttc
     6241 aaccttagga tctctaccct tggcgaaaag tcctctgcca acaatgatga tatctgatcc
     6301 accacttaca acttcgtcga cggttctgta ctgctgaccc aatgcatcgc ctttgtcgtc
     6361 taaacctaca cctggggtca tgattagcca atcaaaccct tcttctcttc ctcccatatc
     6421 gttctgagca atgaacccaa taacgaaatc tttatcactc tttgcaatat caacggtacc
     6481 cttagtatat tcaccgtgtg ctagagaacc cttggaagac aattcagcaa gcatcaataa
     6541 tccccttggt tctttggtga cctcttgcgc accttgtttc aagccagcaa caataccagc
     6601 accagtaacc ccgtgggcgt tggtgatatc agaccattct gcgatacggt aaacgcccga
     6661 tgtatattgt aatttgactg tgttaccgat atcggcgaat tttctgtcct caaatatcaa
     6721 gaacttgtat ttctctgcca atgctttcaa tggaacgaca gtaccctcat aactgaaatc
     6781 atccaagata tcaacgtgtg ttttcaaaag gcaaatgtat ggacccaacg tttcaacaag
     6841 tttcaatagc tcatcagtcg aacgaacgtc aagagaagca cacaaattgg tcttcttttc
     6901 atccattaaa cgtaaaagtt tcgatgcaac cggacttgca tgagtctcag ctctactggt
     6961 atatgatttt gtggacatgt tttatcttcg tttcctgcag gtttttgttc tgtgcagttg
     7021 ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     7081 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     7141 atttaaatga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     7201 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     7261 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     7321 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     7381 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     7441 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     7501 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     7561 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     7621 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     7681 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7741 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7801 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     7861 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7921 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7981 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     8041 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     8101 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     8161 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     8221 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     8281 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     8341 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     8401 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     8461 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     8521 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     8581 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     8641 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     8701 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8761 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     8821 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     8881 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     8941 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     9001 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     9061 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     9121 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     9181 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     9241 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     9301 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     9361 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     9421 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     9481 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     9541 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     9601 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     9661 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     9721 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     9781 gccaagcttg catgcattta aatcattacg accgagattc ccgggtaata actgatataa
     9841 ttaaattgaa gctctaattt gtgagtttag tatacat
//
