LOCUS       sequence-462087-        8396 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid carrying the rad53-5004 allele with its native promoter and 
            terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8396)
  TITLE     van Leeuwen Lab plasmids 2025
REFERENCE   2  (bases 1 to 8396)
  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-rad53-5004
FEATURES             Location/Qualifiers
     source          1..8396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(201..220)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     320..342
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(380..398)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    446..949
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        979..1083
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1084..1152,1153..1944)
                     /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(1302..1321)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2115..2703
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2604..2623
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2857..2874
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2991..3012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3027..3032,3033..3050,3051..3057)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3065..3081
                     /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     3070..3092
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3089..3105
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3089..3105
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3126..3144
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3172..3188
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3173..3189
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6702..6721)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6703..6721)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6728..6745)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6728..6744)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6737..6759)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6885..7340
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6972..6991)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7182..7203
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7600..8262)
                     /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"
     promoter        complement(join(8263..8396,1..54))
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
ORIGIN
        1 aaaaaaaaaa atgaaaatca ttaccgaggc ataaaaaaat atagagtgta ctagaggagg
       61 ccaagagtaa tagaaaaaga aaattgcggg aaaggactgt gttatgactt ccctgactaa
      121 tgccgtgttc aaacgatacc tggcagtgac tcctagcgct caccaagctc ttaaaacggg
      181 aatttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
      241 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
      301 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
      361 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
      421 ataatggttt cttagggtcc ttttcatcac gtgctataaa aataattata atttaaattt
      481 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
      541 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
      601 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
      661 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
      721 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
      781 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
      841 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
      901 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcaggtggc
      961 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     1081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     1141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1321 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     1561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     1621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     1681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     1741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     1801 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     1861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     1921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     1981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     2041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2221 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2881 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2941 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     3001 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     3061 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     3121 ctcgaaatta accctcacta aagggaacaa aagctggtac cgggcccccc ctcgaggtcg
     3181 acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact agtaacgaat
     3241 cgtcgaaact catgtctttt ttatatacgc tttacttaac catagcagaa ctcagcgcac
     3301 aagcccccgg cttacgcggc ggacccgtca actctataat agcgcgctga ggagacaggt
     3361 atctaccatc ttctctctta aaaaggggca gcattttcta tgggtatttg tccttggtta
     3421 cgaaaattgc aaattctcgg ggccttttga ggtttggtcc aattttgccc ttttaacctt
     3481 cttactagga tcaatttgtg attgcgataa acttaccgaa tgtattctct tcaaaatgtt
     3541 ccccgtattc gcattaatcg gtgactctac caagtcgttt agtaccgaat tattgccgtt
     3601 attatttgta tcgctaactg gcggcttctt agccgacatt gatgaagaag aagcctaggg
     3661 ttgatttgca cgttgagctg ccatcatctg ggttaacttt ttcaccaaat ctttttcttc
     3721 ggctgtttgc ttaagtacta ctctttgttc ttgtaacata cctaatccct cgttaaacag
     3781 acctgtagta tcgttaattt ccactttaaa gccaatgaca aatttattgt ttttatccca
     3841 aatgatcttg atttcatctc cgtcttgtaa aaggaatttc gtaccctgta tcatgcggtt
     3901 attatttaaa tagctcacgt tagttccggt gtggcaatac caaatatcat ctaaaccttg
     3961 tgccggagat tcatacatgc ttttgcctac agcatgcctc tttttgaaaa tgaagcaatg
     4021 aactcgagac aacctattgt cttcaatttt acaattgcaa tcctcggatc taccaatgaa
     4081 aaatggattc acaccttgct gaatctccag gctttcttga ataatgctgt caggcaatgg
     4141 ttttaaagtt aaaaacctac cattaccctt tttctttgag ctcttgacat tctcagtatt
     4201 attggaatgt aaaagttttt gttctctagt ttctgcttca atgcttttgg gctgtgtata
     4261 tcgaatagag gcgtgtgcgg gtattttaaa tccttgaatt tttccatctt ggtcttcctg
     4321 atcctgttct tgaagttgtt gctccatttg taatttcctt tgcgctttga caaattcgta
     4381 ttgagcatcg tccatatttt ctaataattt ctgctgcgac aacgattgtg ataaggatat
     4441 ttgtgaaaaa tcaccatatg attgtgagcc caatggactc atcttgatcc agggatgatt
     4501 caaggctttt gcagctgtcg acctattatt tggatccacc tgtaacaatg aatctatgaa
     4561 atctcttgct tcttcagata tccggaaatc tttgaggggc ccttcatgat atgagcctct
     4621 tccaatctgt ttatataatt ggtcctgtgt gctaccacta aaaggtaagt ggcccgttag
     4681 gataacatac acaagacatc ccattgacca catatccact aacgaagagt actcattcct
     4741 ttcttcgtat tcatcaggag atacggatgt atctttacct ctgatgacta caggtgccac
     4801 atatgccaaa gtgccacaga aggttttcat aaaagaccca tttccttgta cttttgccag
     4861 accaaagtcg gttatcttta ccaatacagg atcgtcttgt tcaataagaa tattatcggg
     4921 ctttaggtca cgatggctga tgcccataga gtgaatgtat tttattgctg tgagtatctg
     4981 cctggatatc tccctcccag catcttctcc aaccgcacca tgagcagcaa caaaatccat
     5041 taagtcacca ccagaaacga actccatcac catataataa ctctcagtat cttcataaaa
     5101 tcctttcaat cgtactatcc ttggatgatt gagcttttgc aatacttcta actctcttgt
     5161 cacaccatcc atattgccta ttactttgcg tttacttata atcctcaccg cgaatgtttt
     5221 cccagtagtt ctttcaatgg ctttctttac tgtggcaaat gcaccctggc ccaccacttc
     5281 gtcgataatc gaaaaatcct taaaaatacc agtcttgttg gccaccattg atgatgcagt
     5341 agatgatgta agaccaggag aagctatttt agaggtattt ttcaggttag atcttatgcg
     5401 atcaactttg ttctgctcga ggcactgctt aaatttgtcg tttatgaaaa tgaccagaga
     5461 taaaatatct gattccacgc ctacaccaac ggttatttca tcaccttgag acagtaactg
     5521 attgctgttc ttctcgactt tttgcccatt taaccaggtc ccattagtgg aaatgtcatt
     5581 caataaaagg ttaccgtctt ctcctagtag tatttggaaa tgcttatttg acagtcttga
     5641 aatgtttcct aaatgatagt cacaggctgg gtttctacca aatgtccaaa ctttctttat
     5701 ggatcgtttt tccttaagca cttgtgaaat atcagctgac aaatctcgga tgggaatttg
     5761 acccgtggta caaatgaccc tgcacacaat gttttcgccg atctgttctt gagaaaactt
     5821 ctcaatcaaa aacctttgag tagcctgcgt ggattgctgt gtgggttgtg taatattttc
     5881 catttttagt tgatgttgtg taacactatc ttttctcact attctctctt ttgaagctta
     5941 agaaaatcga atgcatatac aaaattagtt tgtcttccaa taatctctac cgtccttttc
     6001 tgctttctta ccttttcttg atcagagatt tcaaaatggt aactttggta ccctttatct
     6061 tctcaacctg tacctttatt ctataaatgt tttcatttcc aagtctaccc acggtgatga
     6121 tttttctgcc aattttacgc gtcactcccg ttgccgtatc aacgcgtcca caccacctca
     6181 ccctcgaaat gaaaataaat atgttaatag atgcacgttt cattgacatc tgtgacaaac
     6241 actttccggc ttaattaaaa ggaagcaggc tattaatgca ccaaccaaac aaatgatggt
     6301 tcacctcacg ctgaaatctc atctaataaa ggaagaatta ctttggcatg ctttatctcc
     6361 gtcctatagt tgccgatacc acggtagata atacagtatc attctaaaag taccatttag
     6421 ataggtgctg agcgaaccaa taagtaagtt tttgtttttt ctcagtgttt gctttgaatt
     6481 tacattgaaa tacgttgttt tggacgtact acaatagccg ccataataga ttttcgggta
     6541 acttttacac tcttccctct ctttcacgtg agtaagcaca gaataaaatc ctaacggatg
     6601 tgatagaaca aaagggatat caatgacaag agagaggtac tttatgttag tcgcctgtgc
     6661 agaaagaaca gtccactttc cctcagcggc cgctcaattc gccctatagt gagtcgtatt
     6721 acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     6781 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     6841 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg
     6901 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6961 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     7021 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     7081 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     7141 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     7201 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     7261 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     7321 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     7381 tcacaccgca tagatccgtc gagttcaaga gaaaaaaaaa gaaaaagcaa aaagaaaaaa
     7441 ggaaagcgcg cctcgttcag aatgacacgt atagaatgat gcattacctt gtcatcttca
     7501 gtatcatact gttcgtatac atacttactg acattcatag gtatacatat atacacatgt
     7561 atatatatcg tatgctgcag ctttaaataa tcggtgtcac tacataagaa cacctttggt
     7621 ggagggaaca tcgttggtac cattgggcga ggtggcttct cttatggcaa ccgcaagagc
     7681 cttgaacgca ctctcactac ggtgatgatc attcttgcct cgcagacaat caacgtggag
     7741 ggtaattctg ctagcctctg caaagctttc aagaaaatgc gggatcatct cgcaagagag
     7801 atctcctact ttctcccttt gcaaaccaag ttcgacaact gcgtacggcc tgttcgaaag
     7861 atctaccacc gctctggaaa gtgcctcatc caaaggcgca aatcctgatc caaacctttt
     7921 tactccacgc acggccccta gggcctcttt aaaagcttga ccgagagcaa tcccgcagtc
     7981 ttcagtggtg tgatggtcgt ctatgtgtaa gtcaccaatg cactcaacga ttagcgacca
     8041 gccggaatgc ttggccagag catgtatcat atggtccaga aaccctatac ctgtgtggac
     8101 gttaatcact tgcgattgtg tggcctgttc tgctactgct tctgcctctt tttctgggaa
     8161 gatcgagtgc tctatcgcta ggggaccacc ctttaaagag atcgcaatct gaatcttggt
     8221 ttcatttgta atacgcttta ctagggcttt ctgctctgtc atctttgcct tcgtttatct
     8281 tgcctgctca ttttttagta tattcttcga agaaatcaca ttactttata taatgtataa
     8341 ttcattatgt gataatgcca atcgctaaga aaaaaaaaga gtcatccgct aggtgg
//
