LOCUS       sequence-360117-        8969 bp    DNA     circular SYN 15-APR-2025
DEFINITION  YIp for expressing Murine polyomavirus deltaVP1 (GAL1 promoter) and 
            fusion protein VP2C-ERG20(F96W-N127W)-AcNES1 (GAL10 promoter). 
            Contains uracil marker (K. lactis URA3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8969)
  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 8969)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: Linked VLP NES (M)
FEATURES             Location/Qualifiers
     source          1..8969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1149..1590)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(1155..1178)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1473..1590
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(6095..6112)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6095..6111)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6104..6126)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6320..6339)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6439..6461
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6499..6517)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6585..6689
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6690..6758,6759..7550)
                     /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(6908..6927)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7721..8309
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8210..8229
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8463..8480
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8597..8618
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8633..8638,8639..8656,8657..8663)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8671..8687
                     /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     8676..8698
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8695..8711
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8695..8711
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tacagtttta gttttaccga atctatctac atatcttgtc atgtcagggt cacctggaac
       61 tggttcagta ccatcgtaaa ctctaacttc ttcaacttgt gtgttttcac cttccattgg
      121 ttgaccttga acttgtggta acatattatt aaacaaagaa gaaatcaaag aagccattgg
      181 gtatggattt ttaacccatc tttttcttaa agtaatcttg aagtatcttg gcaaacctct
      241 ccaatgatga acatcgtagt ttcttgtaac tctccaaccc ataatatcaa cgcaagataa
      301 gtacaaacct tcacctttac ataatggacc aacaccattt tcatctaaca aaacagttgt
      361 caaagtgtta gtgaattgca aaactggtgg agttgtagta ccaccagtgt aattaccgaa
      421 gtatcttgtg ttttcatttt tagctggatc tggatgccaa atttcaactg ggtacatgcc
      481 atccttatcc aatttagcct ttgaaattgg gttcaaaact tgatccttat taaccatatc
      541 tttcttagtg attgttttaa tagtaacaac accttcttcc ttgtacttag ttcttgcatc
      601 tgtaactaaa ccttgcaaat ccaatggttc accaccaaca gcaaaaacat gatattgtga
      661 accttcaact ggagtagaaa taccctttgt gttaacagta tctgttggct tattaaaacc
      721 atgaacatcc aacaaagaac cagaaccaac aacttctgtt ttaactgaaa ctgcttccca
      781 catttgcaaa gtatcacaag tcaaatcttc gttcaacatt ggtaattgca attttgccat
      841 agaccaagtt ggtaatgtat tattacctgg tgaatcttca gtatcagatg tagccaaatt
      901 aatacctctt gaccaaccgt aatattgacc accttcagtt aaagattctg gtgttggtgg
      961 ttgacccatt cttgggttca aaaatgcttc gatttcagta acagaatctg gacctgtaac
     1021 caaatccaaa acttccatac cacctttaat caataactta ggaactggag ctggtctagg
     1081 gcaagccttt gtgcatttgg tttcgcattt acttacacct gaagccattt tgctagctat
     1141 agttttttct ccttgacgtt aaagtataga ggtatattaa caattttttg ttgatacttt
     1201 tatgacattt gaataagaag taatacaaac cgaaaatgtt gaaagtatta gttaaagtgg
     1261 ttatgcagct tttgcattta tatatctgtt aatagatcaa aaatcatcgc ttcgctgatt
     1321 aattacccca gaaataaggc taaaaaacta atcgcattat tatcctatgg ttgttaattt
     1381 gattcgttga tttgaaggtt tgtggggcca ggttactgcc aatttttcct cttcataacc
     1441 ataaaagcta gtattgtaga atctttattg ttcggagcag tgcggcgcga ggcacatctg
     1501 cgtttcagga acgcgaccgg tgaagaccag gacgcacgga ggagagtctt ccgtcggagg
     1561 gctgtcgccc gctcggcggc ttctaatccg tacttcaata tagcaatgag cagttaagcg
     1621 tattactgaa agttccaaag agaaggtttt tttaggctaa gataatgggg ctctttacat
     1681 ttccacaaca tataagtaag attagatatg gatatgtata tggtggtatt gccatgtaat
     1741 atgattatta aacttctttg cgtccatcca aaaaaaaaag taagaatttt tgaaaattca
     1801 atataaatcg ataaaatggg taatggtggt ccaactccag ctgctcatat tcaagatgaa
     1861 tctggtgaag ttattaaatt ttatcaagct caagttgttt ctcatcaaag agttactcca
     1921 gattggatgt tgccattgat tttgggtttg tatggtgaca tcactccagg tggtggtggt
     1981 tctggtggcg gtggctccgg tggcggcggt tctggatcca tggcttcaga aaaagaaatt
     2041 aggagagaga gattcttgaa cgttttccct aaattagtag aggaattgaa cgcatcgctt
     2101 ttggcttacg gtatgcctaa ggaagcatgt gactggtatg cccactcatt gaactacaac
     2161 actccaggcg gtaagctaaa tagaggtttg tccgttgtgg acacgtatgc tattctctcc
     2221 aacaagaccg ttgaacaatt ggggcaagaa gaatacgaaa aggttgccat tctaggttgg
     2281 tgcattgagt tgttgcaggc ttactggttg gtcgccgatg atatgatgga caagtccatt
     2341 accagaagag gccaaccatg ttggtacaag gttcctgaag ttggggaaat tgccatctgg
     2401 gacgcattca tgttagaggc tgctatctac aagcttttga aatctcactt cagaaacgaa
     2461 aaatactaca tagatatcac cgaattgttc catgaggtca ccttccaaac cgaattgggc
     2521 caattgatgg acttaatcac tgcacctgaa gacaaagtcg acttgagtaa gttctcccta
     2581 aagaagcact ccttcatagt tactttcaag actgcttact attctttcta cttgcctgtc
     2641 gcattggcca tgtacgttgc cggtatcacg gatgaaaagg atttgaaaca agccagagat
     2701 gtcttgattc cattgggtga atacttccaa attcaagatg actacttaga ctgcttcggt
     2761 accccagaac agatcggtaa gatcggtaca gatatccaag ataacaaatg ttcttgggta
     2821 atcaacaagg cattagaact tgcttccgca gaacaaagaa agactttaga cgaaaattac
     2881 ggtaagaagg actcagtcgc agaagccaaa tgcaaaaaga ttttcaatga cttgaaaatt
     2941 gaacagctat accacgaata tgaagagtct attgccaagg atttgaaggc caaaatttct
     3001 caggtcgatg agtctcgtgg cttcaaagct gatgtcttaa ctgcgttctt gaacaaagtt
     3061 tacaagagaa gcaaaagatc tgctggtgga ggaggtactg gtggtgctga gctcatggct
     3121 accgcagcag gtcctatcgc aactaacaac tccccacaaa actccaacgc ttacagaact
     3181 ccaatcgctc cttccgtacc aattactcat aaatggtcta tagctgaaga tttgacatgt
     3241 atttccaatc ctagtaagca caataaccct caaactggtt acagatcatt ttctgacgaa
     3301 ttatacgtta agtacgaaga aaagttggaa gatgttagaa aagcattaag agaagttgaa
     3361 gaaaaccctt tggaaggttt agttatgata gacgctttgc aaagattggg tatcgattac
     3421 catttcagag gtgaaattgg tgcattcttg caaaagcaac aaatcatatc ttcaactcca
     3481 gatggttacc ctgaacatgg tttgtacgaa gtttcaacat tgtttagatt cttaagacaa
     3541 gaaggtcaca atgttaccgc tgacgtcttt aataacttca aggataagga aggtagattc
     3601 agatcagaat tgtcaacaga tattagaggt ttgatgtcct tatacgaagc aagtcaattg
     3661 agaatagaag gtgaagacat cttagatcaa gctgctgatt tctccagtca attgttaggt
     3721 agatggacaa aagatcctaa tcatcacgaa gccagattgg tttctaacac tttaacacat
     3781 ccataccaca agtcattggc tacctttatg ggtcaaaaat tgtcctacat gaactgcaag
     3841 ggtccaaact gggacggtgt cgataatttg caagaattag ctaagatgga tttgactatc
     3901 gtacaaagta tccatcaaaa ggaagtattc caagtttctc aatggtggaa ggatacaggt
     3961 ttagccaatg aattgaaatt ggctagaaac caaccattga agtggtatat gtggcctatg
     4021 gccgctttaa ccgatccaag attcagtgaa gaaagagttg aattaactaa gcctatttct
     4081 tttatataca tcatagatga catcttcgac gtctatggta ccattgaaga attgaccttg
     4141 tttactgatg cagttaatag atgggaattg tctgccgtcg aacaattacc agactacatg
     4201 aaagtatgtt tcaaggcttt gtacgatgtt accaacgaaa tcgcatacaa aatctataaa
     4261 aagcatggtc aaaaccctat tgattccttg caaaagactt gggctagttt gtgcaatgca
     4321 tttttagttg aagcaaagtg gttcgcctct ggtcacttgc caaatgcaga agaatactta
     4381 aagaacggta tcatctcttc aggtgttcat gttgtcttgg cccacatgtt tttcttgtta
     4441 ggtgacggta ttacacaaga atcagttgat ttggtagatg actatccagg tatttccaca
     4501 agtatcgcaa ccattttgag attatctgat gacttgggtt cagccaaaga tgaagaccaa
     4561 gatggttatg atggttctta catcgaatgt tacatgaagg aacataaggg ttccagtgtc
     4621 gattcagcca gagaagaagt aataagaatg atctccgaag catggaaatg tttgaataag
     4681 gaatgcttat caccaaaccc tttttctgaa tcattcagaa taggttcctt gaatatggct
     4741 agaatgatcc ctatgatgta ctcttacgat gacaaccata acttgccaat tttagaagaa
     4801 cacatgaagg caatgatcta taacacatct ttataactcg agatagatta atttaaacag
     4861 tatatgtaca gttttatata tatatatata tatatacata tataaagaaa cctgtgcgtt
     4921 ttttgtattt tcaaatacat ttagttttgg cgccacttct ataaaaggtc tcatgatatt
     4981 gttactctag agatcccaat acaacagatc acgtgatctt tttgtaagat gaagttgaag
     5041 tagtgttgca ccgtgccaat gcaggtggct attagattaa atatgtgatt tgttctatta
     5101 agtttcctgt ataattaatg gggagcgctg attctctttt ggtacgcttc ccatccagca
     5161 tttctgtatc tttcaccttc aaccttagga tctctaccct tggcgaaaag tcctctgcca
     5221 acaatgatga tatctgatcc accacttaca acttcgtcga cggttctgta ctgctgaccc
     5281 aatgcatcgc ctttgtcgtc taaacctaca cctggggtca tgattagcca atcaaaccct
     5341 tcttctcttc ctcccatatc gttctgagca atgaacccaa taacgaaatc tttatcactc
     5401 tttgcaatat caacggtacc cttagtatat tcaccgtgtg ctagagaacc cttggaagac
     5461 aattcagcaa gcatcaataa tccccttggt tctttggtga cctcttgcgc accttgtttc
     5521 aagccagcaa caataccagc accagtaacc ccgtgggcgt tggtgatatc agaccattct
     5581 gcgatacggt aaacgcccga tgtatattgt aatttgactg tgttaccgat atcggcgaat
     5641 tttctgtcct caaatatcaa gaacttgtat ttctctgcca atgctttcaa tggaacgaca
     5701 gtaccctcat aactgaaatc atccaagata tcaacgtgtg ttttcaaaag gcaaatgtat
     5761 ggacccaacg tttcaacaag tttcaatagc tcatcagtcg aacgaacgtc aagagaagca
     5821 cacaaattgg tcttcttttc atccattaaa cgtaaaagtt tcgatgcaac cggacttgca
     5881 tgagtctcag ctctactggt atatgatttt gtggacatgt tttatcttcg tttcctgcag
     5941 gtttttgttc tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta
     6001 catatgcgta tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg
     6061 gagattaccg aatcaaaaaa atttaaatga attcactggc cgtcgtttta caacgtcgtg
     6121 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     6181 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     6241 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     6301 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     6361 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     6421 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     6481 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     6541 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6601 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6661 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     6721 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6781 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     6841 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     6901 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6961 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     7021 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     7081 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     7141 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     7201 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     7261 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7321 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7381 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7441 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7501 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     7561 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7621 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7681 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7741 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7801 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7861 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7921 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7981 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     8041 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     8101 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     8161 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     8221 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     8281 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     8341 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     8401 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     8461 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     8521 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     8581 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     8641 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     8701 acagctatga ccatgattac gccaagcttg catgcattta aatcattacg accgagattc
     8761 ccgggtaata actgatataa ttaaattgaa gctctaattt gtgagtttag tatacatgca
     8821 tttacttata atacagtttt ttagttttgc tggccgcatc ttctcaaata tgcttcccag
     8881 cctgcttttc tgtaacgttc accctctacc ttagcatccc ttccctttgc aaatagtcct
     8941 actagtgcgg ccgcttaatt accagggaa
//
