LOCUS       sequence-462086-        9336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid carrying MSN5 from LY00045 (S288C) with its native promoter 
            and terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9336)
  TITLE     van Leeuwen Lab plasmids 2025
REFERENCE   2  (bases 1 to 9336)
  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"
COMMENT     Sequence Label: pRS313_MSN5-LY00045
FEATURES             Location/Qualifiers
     source          1..9336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        317..504
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             505..1167
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1427..1882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1564..1585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1776..1795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2008..2030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2022..2039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2046..2065
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2046..2064
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6518..6534)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6519..6535)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(6563..6581)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6602..6618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6602..6618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6615..6637)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6626..6642
                     /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(6650..6656,6657..6674,6675..6680))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6695..6716
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6833..6850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7004..7592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7084..7103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7763..8554,8555..8623))
                     /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     8386..8405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8624..8728)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8758..9261
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     9309..9327
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc attatcacat
      421 aatgaattat acattatata aagtaatgtg atttcttcga agaatatact aaaaaatgag
      481 caggcaagat aaacgaaggc aaagatgaca gagcagaaag ccctagtaaa gcgtattaca
      541 aatgaaacca agattcagat tgcgatctct ttaaagggtg gtcccctagc gatagagcac
      601 tcgatcttcc cagaaaaaga ggcagaagca gtagcagaac aggccacaca atcgcaagtg
      661 attaacgtcc acacaggtat agggtttctg gaccatatga tacatgctct ggccaagcat
      721 tccggctggt cgctaatcgt tgagtgcatt ggtgacttac acatagacga ccatcacacc
      781 actgaagact gcgggattgc tctcggtcaa gcttttaaag aggccctagg ggccgtgcgt
      841 ggagtaaaaa ggtttggatc aggatttgcg cctttggatg aggcactttc cagagcggtg
      901 gtagatcttt cgaacaggcc gtacgcagtt gtcgaacttg gtttgcaaag ggagaaagta
      961 ggagatctct cttgcgagat gatcccgcat tttcttgaaa gctttgcaga ggctagcaga
     1021 attaccctcc acgttgattg tctgcgaggc aagaatgatc atcaccgtag tgagagtgcg
     1081 ttcaaggctc ttgcggttgc cataagagaa gccacctcgc ccaatggtac caacgatgtt
     1141 ccctccacca aaggtgttct tatgtagtga caccgattat ttaaagctgc agcatacgat
     1201 atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat acgaacagta
     1261 tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga acgaggcgcg
     1321 ctttcctttt ttctttttgc tttttctttt tttttctctt gaactcgacg gatctatgcg
     1381 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     1441 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1501 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1561 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1621 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1681 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1741 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     1801 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     1861 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2041 aattgtaata cgactcacta tagggcgaat tggagctcca ccgcggtggc ggccgctcta
     2101 gaactagtcg cctcttggtt tttatggtaa tgtactcacc atggcattgt tccttggtca
     2161 ccaatttcta tttaccgggt gatggcggcg cgccacagcg gaggggcatt gccacattgc
     2221 tgcgcgatca aggttggttt ttccaaagag acttcaagag tactaagctt ttgatgtagt
     2281 aatatttgtc aaagagtttt aagagcaaga ttccattgaa cagaggtgct ggagtgacga
     2341 gcgtgtgtgt ttttagccta tctttattcg ttgagcctgt agaagagccg ttcttttaag
     2401 tacaccctga ttagatcccc ctatttgttg caaccgttca agttttgtag atcggaagaa
     2461 gagcaaaaca gaaaaataaa caaggcgaga acgtaaggtg tcattgcatt agccattaaa
     2521 aaccatattc actctttaca ttcgtgtagc aattgttact gtgataaact attgaaatta
     2581 tttatagtat attttgtttt cttcctttta atcacttttt gaatttttag aatctgaaat
     2641 tttttttcta acgttgattg gaagaaaagt aatggattcc acaggcgctt ctcagattgt
     2701 tagcgcacta gatgttattt actcccccaa gtctaacaat tctcaacggc aagaagctca
     2761 gaaattttta gatgaagtga aattatgctc agaatcgccc ttttggggct acgaaattgc
     2821 attgcaaaac cccactaatt ctatattgaa atactttgga ctaggattac tagaccatgc
     2881 agtgaagaaa aactggaatg attatgacga ggggaaaaga gtggccctga gaaaatgggt
     2941 catggaactt aattttgggg tgcaagatta tgataccaga tatattaagg aaaaattagc
     3001 cactttatgg gtagaggtgg caaaaagaac atggggtgag gctcttaagc agacaaaccc
     3061 tacggaagaa cagttactca catcttgggt ggacatggac aataaccttt tcgaactttg
     3121 gaacataaat caatcgtcaa gagaactagc attaataatt ttcagaatcc tttttgaaga
     3181 tgttttcttg ttggacgacc taattgtttt gaaaaggatg acggttattc aacccttatg
     3241 tgtaatgatt gtttgcccta ttgaagtttt cgcaataaaa tacaaattca gtgataaatg
     3301 gaccaaattt aaagctaatg aagagggctg gttttctgtt tggattccag agttgaataa
     3361 tgcattacaa cagaataatt ctgaatacat cataaggctt ttagaaactc tgaagacgtg
     3421 tttaaactgg ccattaacag aggtcattgt aagaaatgac gtgttatcct ccttattgac
     3481 ttgtttatcc agcaatatcc caagggcgca gtcaatggca ttagactcga tacatattct
     3541 gttgacccgt ccctatagta atgaaagcca ttaccaaatg accatagatc gagtttttga
     3601 taatatggac ttattagata gcgtttatga aagcttgttg tttgatccta cagatgatat
     3661 agatgaaacg aaatatccta taattaaaaa gttcgtggat atgatcagtt gtctctatgt
     3721 ttgtgtccca aagattaagg agacaaatgg ccaaattcaa aaatatttta agcttgtatt
     3781 aaagacaact tataacccaa gcttaatcgt tagcggatta acattagacc tatggtgtac
     3841 ctgtcttcga aacgatgaat acttacccaa gctggagaaa tatgtcattc ctgatctact
     3901 acagtttgcc gcggatgctt tagtctatta tgaacagatt gatggtcata tttccaaaaa
     3961 atttgcggag attgattttc aatccaagtc cgaatttcaa acattttgtt ccacttatag
     4021 gaaaagaata agagacatca ttaggttgat ttcttgcgtc gaattagacc ttacttatga
     4081 ttggttgaac aacagattaa acaattattt cagctctcca tttggtcagc aggtcttaag
     4141 ttccacattt ttggaccata aattagagcc ttatttaggt gccctctccc aatacatgat
     4201 tgttgaatgt ttcatcaatg gttgtataag atggaagatc tggtatccaa caggagatga
     4261 ttatgacgaa aaactagatt ccatattaca aaaactggaa attttgtcaa accagttaat
     4321 tgcattaaat ttaagagaac cattgttatt gaagaaacaa atccaaaatt ttgcactttt
     4381 tttgacaatg ttaaaagata atgtgctttt cacactttta gaaaaaatca taacaagcgc
     4441 cacgatggac tatccagaaa ttaatttgga agagagaggc gctgaatcag acgcagttag
     4501 agatttgaga tatgcttgtg gtatcgagtt gaataggatg gctttattaa tgcccgaatc
     4561 tctgaaaaaa atatacccag accttgaaag tgtcattgcc agaatcatgc ccaacttatc
     4621 ttaccatgaa aaaatttcct tcaaatcctt cttattgatc attgtattga agtcttcatt
     4681 agatatgaaa gaggaaagat ttgcggctat tgtggatcca gaacttttag cctggtctga
     4741 taaaacaaca gttgttggtc tttctgattt gcattggttt atggaacgct taggtattgt
     4801 tcagatcgca gaatatttcc agaggcgtga tatcgacgaa aatagtgacc ttttatcaat
     4861 tccaattgat gatgagggta aagagctgaa gtcggaattg accaaacgtt ggcaaagctt
     4921 gtttcctgtt cgtgcaacaa gaatgttcat tcattattca atgcaaagta taaagacaga
     4981 tgaagaattc aaaatgctac aagatctgtg gagacctagg attgtcccaa tattacctta
     5041 catcacgaga ctactgtacc aattgcaatc ctaccacgat ccagataatt ggaaaggatt
     5101 acctacggtt gtgcaatcct tcgtcaagta ttccactatt gaaagatttt gggaggccgg
     5161 agcttcaaat aaatcaaagg atgagtttat tgatgaacat atgaaagcga tgcaaacctt
     5221 acgagacttc gccgactccg taggacacat cattaggtat acaagggaat acactctact
     5281 tgttttaagt gccatatcat cacttggtag tgttttttac cttttggacg aatcgcccga
     5341 cctactactg aactctattg ccatatttaa gccaggtagc aatgagataa gccctggagt
     5401 ttcaacacat ggatggaaac acataatgaa tatagccatt cgtcctatat taaaaggttg
     5461 tccaaaagat tgtctgggga aattcatgcc cgcatttttg cctaaattat ttgaaatcct
     5521 ggacctgctg ctatgtcaga aatggtcctc gcatatgaat gatatggaca tgaacccagt
     5581 tccaactgac gatgatcaaa tgactgaaga aattttggaa gagaatttgt tgagacaact
     5641 aaccacagtt gttgttagaa tagtaattga ttgtgttggt caaggcaatg ccaatccaaa
     5701 cagtgctaaa agcaggctaa ataaccatca aatggagatg agaaaaatta tatttaatga
     5761 tttaaacacc cttgctccat ttttgaaact tttgaaccat ttaatttcct tcaaggatac
     5821 aaaatgttca tttaactcca tcctagtaat gaaatgctgc ttaacctcag tccttaatca
     5881 aaacaacacg gtggatgaat atttcacatt tgaagtgatg aaaaatctgt tgctaaatgt
     5941 gctgtgtaat agtgcgttta aggattcttt ccatgaggca ctatatgctt tcactgtgat
     6001 ttttttgact ctgtgcaaag aatatccctc tgctagagcg ttcctttttg agatatctaa
     6061 tggttataat attgatgagt tatacagaaa tttaaggagc gtggacgaat acaaaacaca
     6121 aagggcactc atgattgatt ttatagattg ggttaaatcc accagtggga aagaagatgg
     6181 taatgtggac catgcaggcg atgagagaaa aagacaagag aaaagggaag ctatcctcaa
     6241 gaaggcaaat gagagattaa ttaagaagaa taaagaaaac ggagacatgc tggatgatcc
     6301 gaacattgaa gacggtgctg tgggtaatct ctttgatgac aactgataat aaggtgccat
     6361 atatacgtag atgtgtagat gaaattagag aaagcaaata gacatctgaa tcgtgctaaa
     6421 aattgtaatt attgtgaaaa aaaaaattct gtattcgcaa tttttttaat gaaagtcggc
     6481 agccggtaaa tatgcaggaa ttcgatatca agcttatcga taccgtcgac ctcgaggggg
     6541 ggcccggtac cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat
     6601 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6661 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6721 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6781 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6841 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6901 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6961 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     7021 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     7081 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     7141 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     7201 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7261 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7321 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7381 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7441 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7501 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7561 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7621 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7681 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7741 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7801 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7861 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7921 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7981 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     8041 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     8101 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     8161 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8221 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8281 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8341 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8401 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8461 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8521 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8581 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8641 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8701 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacggat
     8761 cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat ttgggaattt
     8821 actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa ataaagaagg
     8881 tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa
     8941 agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga tatacattcg
     9001 attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc tacacagaca
     9061 agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa aggtagtatt
     9121 tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat tttttcttta
     9181 atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat ttaaattata
     9241 attattttta tagcacgtga tgaaaaggac cctaagaaac cattattatc atgacattaa
     9301 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
