LOCUS       sequence-209551-        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  JOURNAL   Unpublished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            "Unpublished"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_II
FEATURES             Location/Qualifiers
     source          1..5227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        107..294
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             295..957
                     /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"
     protein_bind    complement(1135..1168)
                     /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(1215..1233)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    1275..1778
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     protein_bind    complement(1797..1830)
                     /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(1915..1935)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1915..1933)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1954..1970)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1954..1970)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1967..1989)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1978..1994
                     /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(2002..2008,2009..2026,2027..2032))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2047..2068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2185..2202)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2356..2944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2436..2455)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3164..3266
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3267..3926
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3333..3352)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4415..4519)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4545..5000)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4682..4703)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4894..4913
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5127..5149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5141..5158
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5142..5158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5168..5187
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5168..5186
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agcgctagga gtcactgcca ggtatcgttt gaacacggca ttagtcaggg aagtcataac
       61 acagtccttt cccgcaattt tctttttcta ttactcttgg cctcctctag tacactctat
      121 atttttttat gcctcggtaa tgattttcat tttttttttt ccacctagcg gatgactctt
      181 tttttttctt agcgattggc attatcacat aatgaattat acattatata aagtaatgtg
      241 atttcttcga agaatatact aaaaaatgag caggcaagat aaacgaaggc aaagatgaca
      301 gagcagaaag ccctagtaaa gcgtattaca aatgaaacca agattcagat tgcgatctct
      361 ttaaagggtg gtcccctagc gatagagcac tcgatcttcc cagaaaaaga ggcagaagca
      421 gtagcagaac aggccacaca atcgcaagtg attaacgtcc acacaggtat agggtttctg
      481 gaccatatga tacatgctct ggccaagcat tccggctggt cgctaatcgt tgagtgcatt
      541 ggtgacttac acatagacga ccatcacacc actgaagact gcgggattgc tctcggtcaa
      601 gcttttaaag aggccctagg ggccgtgcgt ggagtaaaaa ggtttggatc aggatttgcg
      661 cctttggatg aggcactttc cagagcggtg gtagatcttt cgaacaggcc gtacgcagtt
      721 gtcgaacttg gtttgcaaag ggagaaagta ggagatctct cttgcgagat gatcccgcat
      781 tttcttgaaa gctttgcaga ggctagcaga attaccctcc acgttgattg tctgcgaggc
      841 aagaatgatc atcaccgtag tgagagtgcg ttcaaggctc ttgcggttgc cataagagaa
      901 gccacctcgc ccaatggtac caacgatgtt ccctccacca aaggtgttct tatgtagtga
      961 caccgattat ttaaagctgc agcatacgat atatatacat gtgtatatat gtatacctat
     1021 gaatgtcagt aagtatgtat acgaacagta tgatactgaa gatgacaagg taatgcatca
     1081 ttctatacgt gtcattctga acgaggcgcg gctggtaccg ggccccccct cgaaataact
     1141 tcgtatagca tacattatac gaagttatcc atggttgatg aaagaataac gtattctttc
     1201 atcaactagc aggggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     1261 ggtttcttag gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct tttaatgatg
     1321 gaataatttg ggaatttact ctgtgtttat ttatttttat gttttgtatt tggattttag
     1381 aaagtaaata aagaaggtag aagagttacg gaatgaagaa aaaaaaataa acaaaggttt
     1441 aaaaaatttc aacaaaaagc gtactttaca tatatattta ttagacaaga aaagcagatt
     1501 aaatagatat acattcgatt aacgataagt aaaatgtaaa atcacaggat tttcgtgtgt
     1561 ggtcttctac acagacaaga tgaaacaatt cggcattaat acctgagagc aggaagagca
     1621 agataaaagg tagtatttgt tggcgatccc cctagagtct tttacatctt cggaaaacaa
     1681 aaactatttt ttctttaatt tcttttttta ctttctattt ttaatttata tatttatatt
     1741 aaaaaattta aattataatt atttttatag cacgtgatga aaaggaccct gtcgaaataa
     1801 cttcgtatag catacattat acgaagttat ccatggttga tgaaagaata acgtattctt
     1861 tcatcaacta gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta
     1921 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1981 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2041 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2101 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2161 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2221 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2281 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2341 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2401 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2461 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2521 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2581 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2641 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2701 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2761 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2821 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2881 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2941 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3001 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttcgaccg
     3061 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
     3121 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag
     3181 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttgtcga
     3241 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     3301 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     3361 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     3421 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     3481 aattacgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     3541 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     3601 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     3661 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     3721 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     3781 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     3841 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     3901 gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc
     3961 ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat tcgaaagcaa
     4021 attcgacccg gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg tctattgctg
     4081 gtttaccggt ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg
     4141 ccagtttgct caggctctcc ccgtggaggt aataattgac gatatgatcc tttttttctg
     4201 atcaaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     4561 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4621 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4681 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4741 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4801 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4861 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4921 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     4981 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     5041 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     5101 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     5161 gcgcgcgtaa tacgactcac tatagggcga attgggtacg gcgcgcctgc agcatgctcg
     5221 agctagc
//
