LOCUS       sequence-104607-        5861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Kan marker pXR vector uses for both plasmid assembly in vivo, as 
            well as targeted genomic integration..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5861)
  AUTHORS   Chou CC, Patel MT, Gartenberg MR
  TITLE     A series of conditional shuttle vectors for targeted genomic 
            integration in budding yeast.
  JOURNAL   FEMS Yeast Res. 2015 May;15(3):fov010. doi: 10.1093/femsyr/fov010. 
            Epub 2015 Mar 2.
  PUBMED    25736914
REFERENCE   2  (bases 1 to 5861)
  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: "FEMS Yeast 
            Res."; date: "2015-05"; volume: "15(3):fov010. doi"; pages: " 
            10.1093/femsyr/fov010. Epub 2015 Mar 2"
COMMENT     Sequence Label: pXRkMX
FEATURES             Location/Qualifiers
     source          1..5861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..124
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            141..1497
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        141..484
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             485..1294
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(552..571)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1300..1497
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(1589..2044)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1726..1747)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1938..1957
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2021..2401)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2170..2192
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2184..2201
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2185..2201
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2211..2230
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2211..2229
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2238..2345
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2254..2270
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2255..2271
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2305..2321)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2305..2321)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2358..2378)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2358..2376)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2397..2413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2397..2413)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2410..2432)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2421..2437
                     /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(2445..2451,2452..2469,2470..2475))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2490..2511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2628..2645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2799..3387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2879..2898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3558..4349,4350..4418))
                     /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     4181..4200
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4419..4510)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    4545..4578
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5153..5186
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
       61 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
      121 tgtactgaga gtgcaccata gacatggagg cccagaatac cctccttgac agtcttgacg
      181 tgcgcagctc aggggcatga tgtgactgtc gcccgtacat ttagcccata catccccatg
      241 tataatcatt tgcatccata cattttgatg gccgcacggc gcgaagcaaa aattacggct
      301 cctcgctgca gacctgcgag cagggaaacg ctcccctcac agacgcgttg aattgtcccc
      361 acgccgcgcc cctgtagaga aatataaaag gttaggattt gccactgagg ttcttctttc
      421 atatacttcc ttttaaaatc ttgctaggat acagttctca catcacatcc gaacataaac
      481 aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga ttaaattcca acatggatgc
      541 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
      601 tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
      661 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
      721 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
      781 ccccggcaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
      841 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
      901 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
      961 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     1021 aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     1081 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     1141 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     1201 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     1261 gcagtttcat ttgatgctcg atgagttttt ctaatcagta ctgacaataa aaagattctt
     1321 gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct attttaatca
     1381 aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga tgcgaagtta
     1441 agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg
     1501 tcgattcgat actaacgccg ccatccagtg tcgaaaactg cggtgtgaaa taccgcacag
     1561 atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc
     1621 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1681 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     1741 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     1801 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     1861 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1921 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1981 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     2041 gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     2101 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     2161 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact
     2221 cactataggg cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg
     2281 atatcgaatt cctgcagccc gggggatcca ctagttctag agcggccgcc accgcggtgg
     2341 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     2401 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2461 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2521 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2581 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2641 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2701 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2761 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2821 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2881 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2941 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3001 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3061 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3121 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3181 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3241 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3301 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3361 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3421 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3481 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3541 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3601 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3661 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3721 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3781 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3841 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3901 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3961 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     4021 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     4081 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     4141 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     4201 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4261 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4321 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4381 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4441 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4501 aataaacaaa taggggttca gctgaagctt cgacaaccct taatataact tcgtataatg
     4561 tatgctatac gaagttatta ggtctagaga tccccgcgca catttccccg aaaagtgcca
     4621 cctgggtcct tttcatcacg tactataaaa ataattataa tttaaatttt ttaatataaa
     4681 tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc
     4741 cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc
     4801 ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga
     4861 aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct
     4921 tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt
     4981 gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc
     5041 caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta
     5101 aaagatacga ggcgcgtgta agttacaggc aagcgatgcg agaaccctta atataacttc
     5161 gtataatgta tgctatacga agttattagg tgatatcaga tccactagct ataaggttga
     5221 tattctcacg agtaagcggc acttgctact tattgacatt gcagattttt ggctacagaa
     5281 atagtatatt agagattata attgctaatc aaatcaaaat ataaaattag taaaccaaac
     5341 catttatacc cttccttagt agttatggat tgttttttaa tgatatttct gcaaaccaaa
     5401 gaaagattgt tatccagata gaatttagtt ttgatattca tttttttgtt gaagattgaa
     5461 cgccatatct gggcctcata attcaaaaga cggtgccatt atcggtagcg tttcgcattg
     5521 tactggatgg cgcgccttca gaaatttcac agttgatgaa tcgaaaagaa tggtctcatt
     5581 gcaacacgta aggttaagat gtcccttttt accattatag gcaataaatg aatcataaaa
     5641 cgaccgtata ctggtgaaat agtagggaga acgagtacct gtagtaaaaa gtataaatca
     5701 tagttaatcg ggcaatgtcc ctcgatcaag gagtattgtg tcatgttcga gacaaacgcc
     5761 aacatttttg tttcttttgg acaaatgttg tttgcattta tgatccgtta tattttgatc
     5821 taatgtagag ttgcacgtag ttcttactgg caaagaaatc t
//
