LOCUS       sequence-209541-        4781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast shuttle vector with deletable CEN6-ARSH4 cassette in pBIIKS 
            backbone. LoxP site upstream of HIS3. For teaching purposes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4781)
  AUTHORS   Colin Logie
  TITLE     Logie Lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4781)
  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            "Unpublished"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: pBSIIKS_HIS3_I
FEATURES             Location/Qualifiers
     source          1..4781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(120..188,189..980)
                     /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(338..357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1151..1739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1640..1659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1893..1910
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2027..2048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2063..2068,2069..2086,2087..2093)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2101..2117
                     /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     2106..2128
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2125..2141
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2125..2141
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2160..2180
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2162..2180
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(2375..3037)
                     /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(3038..3225)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     protein_bind    complement(3353..3386)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3441..3944
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     3986..4004
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    complement(4015..4048)
                     /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(4128..4147)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4129..4147)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4157..4174)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4157..4173)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4166..4188)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4315..4770
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4402..4421)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4612..4633
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
       61 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      121 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
      181 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
      241 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
      301 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      361 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      421 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      481 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      541 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      601 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      661 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      721 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      781 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      841 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      901 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      961 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1021 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1081 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1261 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1501 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1561 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1621 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1681 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1741 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1801 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1861 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1921 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1981 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     2041 tcactcatta ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa
     2101 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     2161 caattaaccc tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gagctagcag
     2221 ccgcgcctcg ttcagaatga cacgtataga atgatgcatt accttgtcat cttcagtatc
     2281 atactgttcg tatacatact tactgacatt cataggtata catatataca catgtatata
     2341 tatcgtatgc tgcagcttta aataatcggt gtcactacat aagaacacct ttggtggagg
     2401 gaacatcgtt ggtaccattg ggcgaggtgg cttctcttat ggcaaccgca agagccttga
     2461 acgcactctc actacggtga tgatcattct tgcctcgcag acaatcaacg tggagggtaa
     2521 ttctgctagc ctctgcaaag ctttcaagaa aatgcgggat catctcgcaa gagagatctc
     2581 ctactttctc cctttgcaaa ccaagttcga caactgcgta cggcctgttc gaaagatcta
     2641 ccaccgctct ggaaagtgcc tcatccaaag gcgcaaatcc tgatccaaac ctttttactc
     2701 cacgcacggc ccctagggcc tctttaaaag cttgaccgag agcaatcccg cagtcttcag
     2761 tggtgtgatg gtcgtctatg tgtaagtcac caatgcactc aacgattagc gaccagccgg
     2821 aatgcttggc cagagcatgt atcatatggt ccagaaaccc tatacctgtg tggacgttaa
     2881 tcacttgcga ttgtgtggcc tgttctgcta ctgcttctgc ctctttttct gggaagatcg
     2941 agtgctctat cgctagggga ccacccttta aagagatcgc aatctgaatc ttggtttcat
     3001 ttgtaatacg ctttactagg gctttctgct ctgtcatctt tgccttcgtt tatcttgcct
     3061 gctcattttt tagtatattc ttcgaagaaa tcacattact ttatataatg tataattcat
     3121 tatgtgataa tgccaatcgc taagaaaaaa aaagagtcat ccgctaggtg gaaaaaaaaa
     3181 aatgaaaatc attaccgagg cataaaaaaa tatagagtgt actagaggag gccaagagta
     3241 atagaaaaag aaaattgcgg gaaaggactg tgttatgact tccctgacta atgccgtgtt
     3301 caaacgatac ctggcagtga ctcctagcgc tggtaccggg ccccccctcg aaataacttc
     3361 gtatagcata cattatacga agttatccat ggttgatgaa agaataacgt attctttcat
     3421 caactagcag ggtccttttc atcacgtgct ataaaaataa ttataattta aattttttaa
     3481 tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt
     3541 gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc ttttatcttg
     3601 ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt agaagaccac
     3661 acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat atctatttaa
     3721 tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga aattttttaa
     3781 acctttgttt attttttttt cttcattccg taactcttct accttcttta tttactttct
     3841 aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca aattattcca
     3901 tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtcct aagaaaccat
     3961 tattatcatg acattaacct ataaaaatag gcgtatcacg aggcccctgt cgaaataact
     4021 tcgtatagca tacattatac gaagttatcc atggttgatg aaagaataac gtattctttc
     4081 atcaactagt tctagagcgg ccgccaccgc ggtggagctc caattcgccc tatagtgagt
     4141 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     4201 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     4261 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     4321 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4381 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4441 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4501 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4561 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4621 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4681 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4741 attttaacaa aatattaacg cttacaattt aggtggcact t
//
