LOCUS       sequence-209548-        5227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast shuttle vector with deletable CEN6-ARSH4 cassette. For 
            teaching purposes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5227)
  AUTHORS   Colin Logie
  TITLE     Logie Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5227)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBCSK_HIS3_I
FEATURES             Location/Qualifiers
     source          1..5227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..48
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    165..186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(201..206,207..224,225..231)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    239..255
                     /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     244..266
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     263..279
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     263..279
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     298..318
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        300..318
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    403..436
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(447..465)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    507..1010
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     protein_bind    1065..1098
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1226..1413
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             1414..2076
                     /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"
     primer_bind     complement(2273..2292)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2274..2292)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2302..2319)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2302..2318)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2311..2333)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2460..2915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2547..2566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2757..2778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2941..3045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(3534..4193)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4108..4127
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4194..4296)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4516..5104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5005..5024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
       61 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      121 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      181 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      241 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
      301 attaaccctc actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgctctaga
      361 actagttgat gaaagaatac gttattcttt catcaaccat ggataacttc gtataatgta
      421 tgctatacga agttatttcg acaggggcct cgtgatacgc ctatttttat aggttaatgt
      481 catgataata atggtttctt aggacggatc gcttgcctgt aacttacacg cgcctcgtat
      541 cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt atgttttgta
      601 tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag aaaaaaaaat
      661 aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt tattagacaa
      721 gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta aaatcacagg
      781 attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta atacctgaga
      841 gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt cttttacatc
      901 ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat ttttaattta
      961 tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat gaaaaggacc
     1021 ctgctagttg atgaaagaat acgttattct ttcatcaacc atggataact tcgtataatg
     1081 tatgctatac gaagttattt cgaggggggg cccggtacca gcgctaggag tcactgccag
     1141 gtatcgtttg aacacggcat tagtcaggga agtcataaca cagtcctttc ccgcaatttt
     1201 ctttttctat tactcttggc ctcctctagt acactctata tttttttatg cctcggtaat
     1261 gattttcatt tttttttttc cacctagcgg atgactcttt ttttttctta gcgattggca
     1321 ttatcacata atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta
     1381 aaaaatgagc aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag
     1441 cgtattacaa atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg
     1501 atagagcact cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa
     1561 tcgcaagtga ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg
     1621 gccaagcatt ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac
     1681 catcacacca ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctaggg
     1741 gccgtgcgtg gagtaaaaag gtttggatca ggatttgcgc ctttggatga ggcactttcc
     1801 agagcggtgg tagatctttc gaacaggccg tacgcagttg tcgaacttgg tttgcaaagg
     1861 gagaaagtag gagatctctc ttgcgagatg atcccgcatt ttcttgaaag ctttgcagag
     1921 gctagcagaa ttaccctcca cgttgattgt ctgcgaggca agaatgatca tcaccgtagt
     1981 gagagtgcgt tcaaggctct tgcggttgcc ataagagaag ccacctcgcc caatggtacc
     2041 aacgatgttc cctccaccaa aggtgttctt atgtagtgac accgattatt taaagctgca
     2101 gcatacgata tatatacatg tgtatatatg tatacctatg aatgtcagta agtatgtata
     2161 cgaacagtat gatactgaag atgacaaggt aatgcatcat tctatacgtg tcattctgaa
     2221 cgaggcgcgg ctgctagctc gagcatgctg caggcgcgcc gtacccaatt cgccctatag
     2281 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2341 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2401 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     2461 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2521 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2581 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2641 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2701 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2761 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2821 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2881 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     2941 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3001 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3061 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3121 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3181 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3241 atgatgagca ctttttgatc agaaaaaaag gatcatatcg tcaattatta cctccacggg
     3301 gagagcctga gcaaactggc ctcaggcatt tgagaagcac acggtcacac tgcttccggt
     3361 agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga
     3421 accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt
     3481 attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa aaattacgcc
     3541 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     3601 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     3661 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     3721 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     3781 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     3841 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     3901 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     3961 ttcattgcca tacgtaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     4021 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     4081 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     4141 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     4201 tccttagctc ctgaaaatct cgacaactca aaaaatacgc ccggtagtga tcttatttca
     4261 ttatggtgaa agttggaacc tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg
     4321 gcccagggct tcccggtatc aacagggaca ccaggattta tttattctgc gaagtgatct
     4381 tccgtcacag gtatttattc ggtcgaaaag gatctaggtg aagatccttt ttgataatct
     4441 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4501 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4561 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4621 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4681 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4741 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4801 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4861 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4921 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4981 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     5041 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5101 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     5161 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5221 agctgat
//
