LOCUS       sequence-360429-        9877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  YIp for expressing Murine polyomavirus deltaVP1 (GAL1 promoter), 
            VP2C-ERG20(F96C) (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 (G)
FEATURES             Location/Qualifiers
     source          1..9877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4214..4655
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    4214..4331
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     4626..4649
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(6062..6078)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6062..6078)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6075..6097)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6086..6102
                     /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(6110..6116,6117..6134,6135..6140))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6155..6176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6293..6310)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6464..7052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6544..6563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7223..8014,8015..8083))
                     /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     7846..7865
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8084..8188)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8256..8274
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8312..8334)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8434..8453
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8647..8669
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8661..8678
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8662..8678
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atatatatat atatatatat aaaactgtac atatactgtt taaattaatc tatctcgagt
       61 tataaagatg tgttatagat cattgccttc atgtgttctt ctaaaattgg caagttatgg
      121 ttgtcatcgt aagagtacat catagggatc attctagcca tattcaagga acctattctg
      181 aatgattcag aaaaagggtt tggtgataag cattccttat tcaaacattt ccatgcttcg
      241 gagatcattc ttattacttc ttctctggct gaatcgacac tggaaccctt atgttccttc
      301 atgtaacatt cgatgtaaga accatcataa ccatcttggt cttcatcttt ggctgaaccc
      361 aagtcatcag ataatctcaa aatggttgcg atacttgtgg aaatacctgg atagtcatct
      421 accaaatcaa ctgattcttg tgtaataccg tcacctaaca agaaaaacat gtgggccaag
      481 acaacatgaa cacctgaaga gatgataccg ttctttaagt attcttctgc atttggcaag
      541 tgaccagagg cgaaccactt tgcttcaact aaaaatgcat tgcacaaact agcccaagtc
      601 ttttgcaagg aatcaatagg gttttgacca tgctttttat agattttgta tgcgatttcg
      661 ttggtaacat cgtacaaagc cttgaaacat actttcatgt agtctggtaa ttgttcgacg
      721 gcagacaatt cccatctatt aactgcatca gtaaacaagg tcaattcttc aatggtacca
      781 tagacgtcga agatgtcatc tatgatgtat ataaaagaaa taggcttagt taattcaact
      841 ctttcttcac tgaatcttgg atcggttaaa gcggccatag gccacatata ccacttcaat
      901 ggttggtttc tagccaattt caattcattg gctaaacctg tatccttcca ccattgagaa
      961 acttggaata cttccttttg atggatactt tgtacgatag tcaaatccat cttagctaat
     1021 tcttgcaaat tatcgacacc gtcccagttt ggacccttgc agttcatgta ggacaatttt
     1081 tgacccataa aggtagccaa tgacttgtgg tatggatgtg ttaaagtgtt agaaaccaat
     1141 ctggcttcgt gatgattagg atcttttgtc catctaccta acaattgact ggagaaatca
     1201 gcagcttgat ctaagatgtc ttcaccttct attctcaatt gacttgcttc gtataaggac
     1261 atcaaacctc taatatctgt tgacaattct gatctgaatc taccttcctt atccttgaag
     1321 ttattaaaga cgtcagcggt aacattgtga ccttcttgtc ttaagaatct aaacaatgtt
     1381 gaaacttcgt acaaaccatg ttcagggtaa ccatctggag ttgaagatat gatttgttgc
     1441 ttttgcaaga atgcaccaat ttcacctctg aaatggtaat cgatacccaa tctttgcaaa
     1501 gcgtctatca taactaaacc ttccaaaggg ttttcttcaa cttctcttaa tgcttttcta
     1561 acatcttcca acttttcttc gtacttaacg tataattcgt cagaaaatga tctgtaacca
     1621 gtttgagggt tattgtgctt actaggattg gaaatacatg tcaaatcttc agctatagac
     1681 catttatgag taattggtac ggaaggagcg attggagttc tgtaagcgtt ggagttttgt
     1741 ggggagttgt tagttgcgat aggacctgct gcggtagcca tgagctcaga accgccgcca
     1801 ccggagccac cgccaccaga accaccacca cctggagtga tgtcaccata caaacccaaa
     1861 atcaatggca acatccaatc tggagtaact ctttgatgag aaacaacttg agcttgataa
     1921 aatttaataa cttcaccaga ttcatcttga atatgagcag ctggagttgg accaccatta
     1981 cccattttat cgatttttga gggaatattc aactgttttt ttttatcatg ttgatgctct
     2041 gcataataat gcccataaat atttccgacc tgcttttata tctttgctag ccaaactaac
     2101 tgaacatagc tacacattat tttcagcttg gctattttgt gaacactgta tagccagtcc
     2161 ttcggatcac ggtcaacagt tgtccgagcg ctttttggac cctttccctt atttttgggt
     2221 taaggaaaat gacagaaaat atatctaatg agccttcgct caacagtgct ccgaagtata
     2281 gctttccaaa aggagaggca aagcaattta agaatgtatg aacaaaataa aggggaaaaa
     2341 ttaccccctc tactttacca aacgaatact accaataata tttacaactt ttccttatga
     2401 ttttttcact gaagcgcttc gcaatagttg tgagtgatat caaaagtaac gaaatgaact
     2461 ccgcggctcg tgctatattc ttgttgctac cgtccatatc tttccataga ttttcaattt
     2521 ttgatgtctc catggtggta cagagaactt gtaaacaatt cggtccctac atgtgaggaa
     2581 attcgctgtg acacttttat cactgaactc caaatttaaa aaatagcata aaattcgtta
     2641 tacagcaaat ctatgtgttg caattaagaa ctaaaagata tagagtgcat attttcaaga
     2701 aggatagtaa gctggcaaaa gatctttatt tgcttctctt gtaaactttg ttcaaaaacg
     2761 cagttaagac atcagctttg aagccacgag actcatcgac ctgagaaatt ttagccttca
     2821 aatccttggc aatagactct tcatattcgt ggtatagctg ttcaattttc aagtcattga
     2881 aaatcttttt gcatttggct tctgcgactg agtccttctt accgtaattt tcgtctaaag
     2941 tctttctttg ttctgcggaa gcaagttcta atgccttgtt gattacccaa gaacatttgt
     3001 tatcttggat atctgtaccg atcttaccga tctgttctgg ggtaccgaag cagtctaagt
     3061 agtcatcttg aatttggaag tattcaccca atggaatcaa gacatctctg gcttgtttca
     3121 aatccttttc atccgtgata ccggcaacgt acatggccaa tgcgacaggc aagtagaaag
     3181 aatagtaagc agtcttgaaa gtaactatga aggagtgctt ctttagggag aacttactca
     3241 agtcgacttt gtcttcaggt gcagtgatta agtccatcaa ttggcccaat tcggtttgga
     3301 aggtgacctc atggaacaat tcggtgatat ctatgtagta tttttcgttt ctgaagtgag
     3361 atttcaaaag cttgtagata gcagcctcta acatgaatgc gtcattgatg gcaatttccc
     3421 caacttcagg aaccttgtac caacatggtt ggcctcttct ggtaatggac ttgtccatca
     3481 tatcatcggc gaccaaacag taagcctgca acaactcaat gcaccaacct agaatggcaa
     3541 ccttttcgta ttcttcttgc cccaattgtt caacggtctt gttggagaga atagcatacg
     3601 tgtccacaac ggacaaacct ctatttagct taccgcctgg agtgttgtag ttcaatgagt
     3661 gggcatacca gtcacatgct tccttaggca taccgtaagc caaaagcgat gcgttcaatt
     3721 cctctactaa tttagggaaa acgttcaaga atctctctct cctaatttct ttttctgaag
     3781 ccatggatcc agaaccgccg ccaccggagc caccgccacc agaaccacca ccacctggag
     3841 tgatgtcacc atacaaaccc aaaatcaatg gcaacatcca atctggagta actctttgat
     3901 gagaaacaac ttgagcttga taaaatttaa taacttcacc agattcatct tgaatatgag
     3961 cagctggagt tggaccacca ttacccattt tatcgattta tattgaattt tcaaaaattc
     4021 ttactttttt ttttggatgg acgcaaagaa gtttaataat catattacat ggcaatacca
     4081 ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa gagccccatt
     4141 atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa ctgctcattg
     4201 ctatattgaa gtacggatta gaagccgccg agcgggcgac agccctccga cggaagactc
     4261 tcctccgtgc gtcctggtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc
     4321 gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa
     4381 attggcagta acctggcccc acaaaccttc aaatcaacga atcaaattaa caaccatagg
     4441 ataataatgc gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga
     4501 tttttgatct attaacagat atataaatgc aaaagctgca taaccacttt aactaatact
     4561 ttcaacattt tcggtttgta ttacttctta ttcaaatgtc ataaaagtat caacaaaaaa
     4621 ttgttaatat acctctatac tttaacgtca aggagaaaaa actatagcta gcaaaatggc
     4681 ttcaggtgta agtaaatgcg aaaccaaatg cacaaaggct tgccctagac cagctccagt
     4741 tcctaagtta ttgattaaag gtggtatgga agttttggat ttggttacag gtccagattc
     4801 tgttactgaa atcgaagcat ttttgaaccc aagaatgggt caaccaccaa caccagaatc
     4861 tttaactgaa ggtggtcaat attacggttg gtcaagaggt attaatttgg ctacatctga
     4921 tactgaagat tcaccaggta ataatacatt accaacttgg tctatggcaa aattgcaatt
     4981 accaatgttg aacgaagatt tgacttgtga tactttgcaa atgtgggaag cagtttcagt
     5041 taaaacagaa gttgttggtt ctggttcttt gttggatgtt catggtttta ataagccaac
     5101 agatactgtt aacacaaagg gtatttctac tccagttgaa ggttcacaat atcatgtttt
     5161 tgctgttggt ggtgaaccat tggatttgca aggtttagtt acagatgcaa gaactaagta
     5221 caaggaagaa ggtgttgtta ctattaaaac aatcactaag aaagatatgg ttaataagga
     5281 tcaagttttg aacccaattt caaaggctaa attggataag gatggcatgt acccagttga
     5341 aatttggcat ccagatccag ctaaaaatga aaacacaaga tacttcggta attacactgg
     5401 tggtactaca actccaccag ttttgcaatt cactaacact ttgacaactg ttttgttaga
     5461 tgaaaatggt gttggtccat tatgtaaagg tgaaggtttg tacttatctt gcgttgatat
     5521 tatgggttgg agagttacaa gaaactacga tgttcatcat tggagaggtt tgccaagata
     5581 cttcaagatt actttaagaa aaagatgggt taaaaatcca tacccaatgg cttctttgat
     5641 ttcttctttg tttaataata tgttaccaca agttcaaggt caaccaatgg aaggtgaaaa
     5701 cacacaagtt gaagaagtta gagtttacga tggtactgaa ccagttccag gtgaccctga
     5761 catgacaaga tatgtagata gattcggtaa aactaaaact gtattccctg gtaattaagc
     5821 ggccgcacta gtaggactat ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta
     5881 cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact aaaaaactgt
     5941 attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt aattatatca
     6001 gttattaccc gggaatctcg gtcgtaatga tttaaatgca tgcaagcttg gcgtaatcat
     6061 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6121 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6181 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6241 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6301 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6361 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6421 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6481 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6541 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6601 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6661 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6721 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6781 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6841 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6901 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6961 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7021 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7081 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7141 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7201 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7261 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7321 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7381 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7441 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7501 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7561 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7621 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7681 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7741 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7801 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7861 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7921 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7981 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8041 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8101 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8161 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8221 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8281 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     8341 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     8401 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     8461 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     8521 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     8581 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     8641 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcatttaa atttttttga
     8701 ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata
     8761 tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca
     8821 cagaacaaaa acctgcagga aacgaagata aaacatgtcc acaaaatcat ataccagtag
     8881 agctgagact catgcaagtc cggttgcatc gaaactttta cgtttaatgg atgaaaagaa
     8941 gaccaatttg tgtgcttctc ttgacgttcg ttcgactgat gagctattga aacttgttga
     9001 aacgttgggt ccatacattt gccttttgaa aacacacgtt gatatcttgg atgatttcag
     9061 ttatgagggt actgtcgttc cattgaaagc attggcagag aaatacaagt tcttgatatt
     9121 tgaggacaga aaattcgccg atatcggtaa cacagtcaaa ttacaatata catcgggcgt
     9181 ttaccgtatc gcagaatggt ctgatatcac caacgcccac ggggttactg gtgctggtat
     9241 tgttgctggc ttgaaacaag gtgcgcaaga ggtcaccaaa gaaccaaggg gattattgat
     9301 gcttgctgaa ttgtcttcca agggttctct agcacacggt gaatatacta agggtaccgt
     9361 tgatattgca aagagtgata aagatttcgt tattgggttc attgctcaga acgatatggg
     9421 aggaagagaa gaagggtttg attggctaat catgacccca ggtgtaggtt tagacgacaa
     9481 aggcgatgca ttgggtcagc agtacagaac cgtcgacgaa gttgtaagtg gtggatcaga
     9541 tatcatcatt gttggcagag gacttttcgc caagggtaga gatcctaagg ttgaaggtga
     9601 aagatacaga aatgctggat gggaagcgta ccaaaagaga atcagcgctc cccattaatt
     9661 atacaggaaa cttaatagaa caaatcacat atttaatcta atagccacct gcattggcac
     9721 ggtgcaacac tacttcaact tcatcttaca aaaagatcac gtgatctgtt gtattgggat
     9781 ctctagagta acaatatcat gagacctttt atagaagtgg cgccaaaact aaatgtattt
     9841 gaaaatacaa aaaacgcaca ggtttcttta tatatgt
//
