LOCUS       sequence-462290-        9332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid carrying MSN5 from LY00018 (YJM975) with its native promoter
            and terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9332)
  TITLE     van Leeuwen Lab plasmids 2025
REFERENCE   2  (bases 1 to 9332)
  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-LY00018
FEATURES             Location/Qualifiers
     source          1..9332
                     /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(6514..6530)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6515..6531)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(6559..6577)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6598..6614)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6598..6614)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6611..6633)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6622..6638
                     /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(6646..6652,6653..6670,6671..6676))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6691..6712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6829..6846)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7000..7588)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7080..7099)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7759..8550,8551..8619))
                     /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     8382..8401
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8620..8724)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    8754..9257
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     9305..9323
                     /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 tctttgttga gcctgtagaa gagccgttct tttaagtaca
     2401 ccctgattag atccccctat ttgttgcaac cgttcaagtt ttgtagatcg gaagaagagc
     2461 aaaacagaaa aataaacaag gcgagaacgt aaggtgtcat tgcattagcc attaaaaacc
     2521 atattcactc tttacattcg tgtagcaatt gttactgtga taaactattg aaattattta
     2581 tagtatattt tgttttcttc cttttaatca ctttttgaat ttttagaatc tgaaattttt
     2641 tttctaacgt tgattggaag aaaagtaatg gattccacag gcgcttctca gattgttagc
     2701 gcactagatg ttatttactc ccccaagtct aacaattctc aacggcaaga agctcagaaa
     2761 tttttagatg aagtgaaatt atgctcagaa tcgccctttt ggggctacga aattgcattg
     2821 caaaacccca ctaattctat attgaaatac tttggactag gattactaga ccatgcagtg
     2881 aagaaaaact ggaatgatta tgacgagggg aaaagagtgg ccctgagaaa atgggtcatg
     2941 gaacttaatt ttggggtgca agattatgat accagatata ttaaggaaaa attagccact
     3001 ttatgggtag aggtggcaaa aagaacatgg ggtgaggctc ttaagcagac aaaccctacg
     3061 gaagaacagt tactcacatc ttgggtggac atggacaata accttttcga actttggaac
     3121 ataaatcaat cgtcaagaga actagcatta ataattttca gaatcctttt tgaagatgtt
     3181 ttcttgttgg acgacctaat tgttttgaaa aggatgacgg ttattcaacc cttatgtgta
     3241 atgattgttt gccctattga agttttcgca ataaaataca aattcagtga taaatggacc
     3301 aaatttaaag ctaatgaaga gggctggttt tctgtttgga ttccagagtt gaataatgca
     3361 ttacaacaga ataattctga atacatcata aggcttttag aaactctgaa gacgtgttta
     3421 aactggccat taacagaggt cattgtaaga aatgacgtgt tatcctcctt attgacttgt
     3481 ttatccagca atatcccaag ggcgcagtca atggcattag actcgataca tattctgttg
     3541 acccgtccct atagtaatga aagccattac caaatgacca tagatcgagt ttttgataat
     3601 atggacttat tagatagcgt ttatgaaagc ttgttgtttg atcctacaga tgatatagat
     3661 gaaacgaaat atcctataat taaaaagttc gtggatatga tcagttgtct ctatgtttgt
     3721 gtcccaaaga ttaaggagac aaatggccaa attcaaaaat attttaagct tgtattaaag
     3781 acaacttata acccaagctt aatcgttagc ggattaacat tagacctatg gtgtacctgt
     3841 cttcgaaacg atgaatactt acccaagctg gagaaatatg tcattcctga tctactacag
     3901 tttgccgcgg atgctttagt ctattatgaa cagattgatg gtcatatttc caaaaaattt
     3961 gcggagattg attttcaatc caagtccgaa tttcaaacat tttgttccac ttataggaaa
     4021 agaataagag acatcattag gttgatttct tgcgtcgaat tagaccttac ttatgattgg
     4081 ttgaacaaca gattaaacaa ttatttcagc tctccatttg gtcagcaggt cttaagttcc
     4141 acatttttgg accataaatt agagccttat ttaggtgccc tctcccaata catgattgtt
     4201 gaatgtttca tcaatggttg tataagatag aagatctggt atccaacagg agatgattat
     4261 gacgaaaaac tagattccat attacaaaaa ctggaaattt tgtcaaacca gttaattgca
     4321 ttaaatttaa gagaaccatt gttattgaag aaacaaatcc aaaattttgc actttttttg
     4381 acaatgttaa aagataatgt gcttttcaca cttttagaaa aaatcataac aagcgccacg
     4441 atggactatc cagaaattaa tttggaagag agaggcgctg aatcagacgc agttagagat
     4501 ttgagatatg cttgtggtat cgagttgaat aggatggctt tattaatgcc cgaatctctg
     4561 aaaaaaatat acccagacct tgaaagtgtc attgccagaa tcatgcccaa cttatcttac
     4621 catgaaaaaa tttccttcaa atccttctta ttgatcattg tattgaagtc ttcattagat
     4681 atgaaagagg aaagatttgc ggctattgtg gatccagaac ttttagcctg gtctgataaa
     4741 acaacagttg ttggtctttc tgatttgcat tggtttatgg aacgcttagg tattgttcag
     4801 atcgcagaat atttccagag gcgtgatatc gacgaaaata gtgacctttt atcaattcca
     4861 attgatgatg agggtaaaga gctgaagtcg gaattgacca aacgttggca aagcttgttt
     4921 cctgttcgtg caacaagaat gttcattcat tattcaatgc aaagtataaa gacagatgaa
     4981 gaattcaaaa tgctacaaga tctgtggaga cctaggattg tcccaatatt accttacatc
     5041 acgagactac tgtaccaatt gcaatcctac cacgatccag ataattggaa aggattacct
     5101 acggttgtgc aatccttcgt caagtattcc actattgaaa gattttggga ggccggagct
     5161 tcaaataaat caaaggatga gtttattgat gaacatatga aagcgatgca aaccttacga
     5221 gacttcgccg actccgtagg acacatcatt aggtatacaa gggaatacac tctacttgtt
     5281 ttaagtgcca tatcatcact tggtagtgtt ttttaccttt tggacgaatc gcccgaccta
     5341 ctactgaact ctattgccat atttaagcca ggtagcaatg agataagccc tggagtttca
     5401 acacatggat ggaaacacat aatgaatata gccattcgtc ctatattaaa aggttgtcca
     5461 aaagattgtc tggggaaatt catgcccgca tttttgccta aattatttga aatcctggac
     5521 ctgctgctat gtcagaaatg gtcctcgcat atgaatgata tggacatgaa cccagttcca
     5581 actgacgatg atcaaatgac tgaagaaatt ttggaagaga atttgttgag acaactaacc
     5641 acagttgttg ttagaatagt aattgattgt gttggtcaag gcaatgccaa tccaaacagt
     5701 gctaaaagca ggctaaataa ccatcaaatg gagatgagaa aaattatatt taatgattta
     5761 aacacccttg ctccattttt gaaacttttg aaccatttaa tttccttcaa ggatacaaaa
     5821 tgttcattta actccatcct agtaatgaaa tgctgcttaa cctcagtcct taatcaaaac
     5881 aacacggtgg atgaatattt cacatttgaa gtgatgaaaa atctgttgct aaatgtgctg
     5941 tgtaatagtg cgtttaagga ttctttccat gaggcactat atgttttcac tgtgattttt
     6001 ttgactctgt gcaaagaata tccctctgct agagcgttcc tttttgagat atctaatggt
     6061 tatcatattg atgagttata cagaaattta aggagcgtgg acgaatacaa aacacaaagg
     6121 gcactcatga ttgattttat agattgggtt aaatccacca gtgggaaaga agatggtaat
     6181 gtggaccatg caggcgatga gagaaaaaga caagagaaaa gggaagctat cctcaagaag
     6241 gcaaatgaga gattaattaa gaagaataaa gaaaacggag acatgctgga tgatccgaac
     6301 attgaagacg gtgctgtggg taatctcttt gatgacaact gataataagg tgccatatat
     6361 acgtagatgt gtagatgaaa ttagagaaag caaatagaca tctgaatcgt gctaaaaatt
     6421 gtaattattg tgaaaaaaaa aattctgtat tcgcaatttt tttaatgaaa gtcggcagcc
     6481 ggtaaatatg caggaattcg atatcaagct tatcgatacc gtcgacctcg agggggggcc
     6541 cggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc
     6601 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6661 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6721 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6781 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6841 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6901 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6961 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7021 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7081 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7141 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7201 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7261 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7321 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7381 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7441 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7501 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7561 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7621 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7681 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7741 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7801 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7861 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7921 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7981 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8041 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8101 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8161 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8221 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8281 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8341 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8401 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8461 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8521 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8581 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8641 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8701 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acggatcgct
     8761 tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg gaatttactc
     8821 tgtgtttatt tatttttatg ttttgtattt ggattttaga aagtaaataa agaaggtaga
     8881 agagttacgg aatgaagaaa aaaaaataaa caaaggttta aaaaatttca acaaaaagcg
     8941 tactttacat atatatttat tagacaagaa aagcagatta aatagatata cattcgatta
     9001 acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat
     9061 gaaacaattc ggcattaata cctgagagca ggaagagcaa gataaaaggt agtatttgtt
     9121 ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa aactattttt tctttaattt
     9181 ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa attataatta
     9241 tttttatagc acgtgatgaa aaggacccta agaaaccatt attatcatga cattaaccta
     9301 taaaaatagg cgtatcacga ggccctttcg tc
//
