LOCUS       sequence-205852-        5225 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 5225)
  AUTHORS   Colin Logie
  TITLE     Logie Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5225)
  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            "Unpublished"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..5225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..129
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(617..1276)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1191..1210
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1277..1379)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      1599..2187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2088..2107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2341..2358
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2475..2496
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2511..2516,2517..2534,2535..2541)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2549..2565
                     /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     2554..2576
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2573..2589
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2573..2589
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2608..2628
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2610..2628
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2713..2746
                     /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(2757..2775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2818..3321
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     protein_bind    3376..3409
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3537..3724
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3725..4384
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     primer_bind     complement(4581..4600)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4582..4600)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4610..4627)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4610..4626)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4619..4641)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4768..5223
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4855..4874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5065..5086
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac ccttgataaa tgcttcaata atattgaaaa
      121 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      181 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      241 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      301 tttcgccccg aagaacgttt tccaatgatg agcacttttg atcagaaaaa aaggatcata
      361 tcgtcaatta ttacctccac ggggagagcc tgagcaaact ggcctcaggc atttgagaag
      421 cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg
      481 gctatttaac gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc
      541 cattcatccg cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat
      601 aactgcctta aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
      661 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
      721 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
      781 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
      841 agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
      901 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
      961 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     1021 cccatatcac cagctcaccg tctttcattg ccatacgtaa ttccggatga gcattcatca
     1081 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
     1141 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
     1201 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     1261 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgacaac tcaaaaaata
     1321 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     1381 cgtctcattt tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat
     1441 ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggtcgaa aaggatctag
     1501 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     1561 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1621 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1681 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1741 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1801 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1861 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1921 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1981 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2041 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2101 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2161 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2221 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2281 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2341 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     2401 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     2461 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     2521 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     2581 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
     2641 gagctccacc gcggtggcgg ccgctctaga actagttgat gaaagaatac gttattcttt
     2701 catcaaccat ggataacttc gtataatgta tgctatacga agttatttcg acaggggcct
     2761 cgtgatacgc ctatttttat aggttaatgt catggataat aatggtttct taggacggat
     2821 cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat ttgggaattt
     2881 actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa ataaagaagg
     2941 tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat ttcaacaaaa
     3001 agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga tatacattcg
     3061 attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc tacacagaca
     3121 agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa aggtagtatt
     3181 tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat tttttcttta
     3241 atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat ttaaattata
     3301 attattttta tagcacgtga tgaaaaggac cctgctagtt gatgaaagaa tacgttattc
     3361 tttcatcaac catggataac ttcgtataat gtatgctata cgaagttatt tcgagggggg
     3421 gcccggtacc agcgctagga gtcactgcca ggtatcgttt gaacacggca ttagtcaggg
     3481 aagtcataac acagtccttt cccgcaattt tctttttcta ttactcttgg cctcctctag
     3541 tacactctat atttttttat gcctcggtaa tgattttcat tttttttttt ccacctagcg
     3601 gatgactctt tttttttctt agcgattggc attatcacat aatgaattat acattatata
     3661 aagtaatgtg atttcttcga agaatatact aaaaaatgag caggcaagat aaacgaaggc
     3721 aaagatgaca gagcagaaag ccctagtaaa gcgtattaca aatgaaacca agattcagat
     3781 tgcgatctct ttaaagggtg gtcccctagc gatagagcac tcgatcttcc cagaaaaaga
     3841 ggcagaagca gtagcagaac aggccacaca atcgcaagtg attaacgtcc acacaggtat
     3901 agggtttctg gaccatatga tacatgctct ggccaagcat tccggctggt cgctaatcgt
     3961 tgagtgcatt ggtgacttac acatagacga ccatcacacc actgaagact gcgggattgc
     4021 tctcggtcaa gcttttaaag aggccctact ggcgcgtgga gtaaaaaggt ttggatcagg
     4081 atttgcgcct ttggatgagg cactttccag agcggtggta gatctttcga acaggccgta
     4141 cgcagttgtc gaacttggtt tgcaaaggga gaaagtagga gatctctctt gcgagatgat
     4201 cccgcatttt cttgaaagct ttgcagaggc tagcagaatt accctccacg ttgattgtct
     4261 gcgaggcaag aatgatcatc accgtagtga gagtgcgttc aaggctcttg cggttgccat
     4321 aagagaagcc acctcgccca atggtaccaa cgatgttccc tccaccaaag gtgttcttat
     4381 gtagtgacac cgattattta aagctgcagc atacgatata tatacatgtg tatatatgta
     4441 tacctatgaa tgtcagtaag tatgtatacg aacagtatga tactgaagat gacaaggtaa
     4501 tgcatcattc tatacgtgtc attctgaacg aggcgcggct gctagctcga gcatgctgca
     4561 ggcgcgccgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     4621 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4681 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4741 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     4801 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4861 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4921 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4981 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5041 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5101 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5161 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5221 tttag
//
