LOCUS       sequence-108401-        6693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Leu2 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 6693)
  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 6693)
  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: pXRL2
FEATURES             Location/Qualifiers
     source          1..6693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(619..1713)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1714..2121)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2422..2877)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2559..2580)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2771..2790
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2854..3234)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3003..3025
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3017..3034
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3018..3034
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3044..3063
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3044..3062
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3071..3178
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3087..3103
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3088..3104
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3138..3154)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3138..3154)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3191..3211)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3191..3209)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3230..3246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3230..3246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3243..3265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3254..3270
                     /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(3278..3284,3285..3302,3303..3308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3323..3344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3461..3478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3632..4220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3712..3731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4391..5182,5183..5251))
                     /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     5014..5033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5252..5343)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    5378..5411
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5986..6019
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
       61 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
      121 ttgtactgag agtgcaccat atcgactacg tcgttaaggc cgtttctgac agagtaaaat
      181 tcttgaggga actttcacca ttatgggaaa tggttcaaga aggtattgac ttaaactcca
      241 tcaaatggtc aggtcattga gtgtttttta tttgttgtat tttttttttt tagagaaaat
      301 cctccaatat ataaattagg aatcatagtt tcatgatttt ctgttacacc taactttttg
      361 tgtggtgccc tcctccttgt caatattaat gttaaagtgc aattcttttt ccttatcacg
      421 ttgagccatt agtatcaatt tgcttacctg tattccttta ctatcctcct ttttctcctt
      481 cttgataaat gtatgtagat tgcgtatata gtttcgtcta ccctatgaac atattccatt
      541 ttgtaatttc gtgtcgtttc tattatgaat ttcatttata aagtttatgt acaaatatca
      601 taaaaaaaga gaatcttttt aagcaaggat tttcttaact tcttcggcga cagcatcacc
      661 gacttcggtg gtactgttgg aaccacctaa atcaccagtt ctgatacctg catccaaaac
      721 ctttttaact gcatcttcaa tggccttacc ttcttcaggc aagttcaatg acaatttcaa
      781 catcattgca gcagacaaga tagtggcgat agggtcaacc ttattctttg gcaaatctgg
      841 agcagaaccg tggcatggtt cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc
      901 caaggacgca gatggcaaca aacccaagga acctgggata acggaggctt catcggagat
      961 gatatcacca aacatgttgc tggtgattat aataccattt aggtgggttg ggttcttaac
     1021 taggatcatg gcggcagaat caatcaattg atgttgaacc ttcaatgtag ggaattcgtt
     1081 cttgatggtt tcctccacag tttttctcca taatcttgaa gaggccaaaa cattagcttt
     1141 atccaaggac caaataggca atggtggctc atgttgtagg gccatgaaag cggccattct
     1201 tgtgattctt tgcacttctg gaacggtgta ttgttcacta tcccaagcga caccatcacc
     1261 atcgtcttcc tttctcttac caaagtaaat acctcccact aattctctga caacaacgaa
     1321 gtcagtacct ttagcaaatt gtggcttgat tggagataag tctaaaagag agtcggatgc
     1381 aaagttacat ggtcttaagt tggcgtacaa ttgaagttct ttacggattt ttagtaaacc
     1441 ttgttcaggt ctaacactac cggtacccca tttaggacca cccacagcac ctaacaaaac
     1501 ggcatcaacc ttcttggagg cttccagcgc ctcatctgga agtgggacac ctgtagcatc
     1561 gatagcagca ccaccaatta aatgattttc gaaatcgaac ttgacattgg aacgaacatc
     1621 agaaatagct ttaagaacct taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc
     1681 tggcaaaacg acgatcttct taggggcaga cattagaatg gtatatcctt gaaatatata
     1741 tatatatatt gctgaaatgt aaaaggtaag aaaagttaga aagtaagacg attgctaacc
     1801 acctattgga aaaaacaata ggtccttaaa taatattgtc aacttcaagt attgtgatgc
     1861 aagcatttag tcatgaacgc ttctctattc tatatgaaaa gccggttccg gcgctctcac
     1921 ctttcctttt tctcccaatt tttcagttga aaaaggtata tgcgtcaggc gacctctgaa
     1981 attaacaaaa aatttccagt catcgaattt gattctgtgc gatagcgccc ctgtgtgttc
     2041 tcgttatgtt gaggaaaaaa ataatggttg ctaagagatt cgaactcttg catcttacga
     2101 tacctgagta ttcccacagt taactgcggt caagatattt cttgaatcag gcgccttaga
     2161 ccgctcggcc aaacaaccaa ttacttgttg agaaatagag tataattatc ctataaatat
     2221 aacgtttttg aacacacatg aacaaggaag tacaggacaa ttgattttga agagaatgtg
     2281 gattttgatg taattgttgg gattccattt ttaataaggc aataatatta ggtatgtaga
     2341 tatactagaa gttctcctcg accggtcgat atgcggtgtg aaataccgca cagatgcgta
     2401 aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2461 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2521 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2581 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2641 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2701 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2761 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2821 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca
     2881 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     2941 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     3001 ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata
     3061 gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga
     3121 attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg tggagctcca
     3181 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     3241 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3301 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3361 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3421 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3481 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3541 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3601 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3661 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3721 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3781 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3841 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3901 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3961 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4021 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4081 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4141 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4201 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4261 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4321 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4381 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4441 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4501 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4561 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4621 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4681 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4741 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4801 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4861 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4921 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4981 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5041 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5101 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5161 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5221 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5281 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5341 aaataggggt tcagctgaag cttcgacaac ccttaatata acttcgtata atgtatgcta
     5401 tacgaagtta ttaggtctag agatccccgc gcacatttcc ccgaaaagtg ccacctgggt
     5461 ccttttcatc acgtactata aaaataatta taatttaaat tttttaatat aaatatataa
     5521 attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat
     5581 gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc ttcctgctct
     5641 caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca cgaaaatcct
     5701 gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg
     5761 tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt
     5821 tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac
     5881 aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata
     5941 cgaggcgcgt gtaagttaca ggcaagcgat gcgagaaccc ttaatataac ttcgtataat
     6001 gtatgctata cgaagttatt aggtgatatc agatccacta gctataaggt tgatattctc
     6061 acgagtaagc ggcacttgct acttattgac attgcagatt tttggctaca gaaatagtat
     6121 attagagatt ataattgcta atcaaatcaa aatataaaat tagtaaacca aaccatttat
     6181 acccttcctt agtagttatg gattgttttt taatgatatt tctgcaaacc aaagaaagat
     6241 tgttatccag atagaattta gttttgatat tcattttttt gttgaagatt gaacgccata
     6301 tctgggcctc ataattcaaa agacggtgcc attatcggta gcgtttcgca ttgtactgga
     6361 tggcgcgcct tcagaaattt cacagttgat gaatcgaaaa gaatggtctc attgcaacac
     6421 gtaaggttaa gatgtccctt tttaccatta taggcaataa atgaatcata aaacgaccgt
     6481 atactggtga aatagtaggg agaacgagta cctgtagtaa aaagtataaa tcatagttaa
     6541 tcgggcaatg tccctcgatc aaggagtatt gtgtcatgtt cgagacaaac gccaacattt
     6601 ttgtttcttt tggacaaatg ttgtttgcat ttatgatccg ttatattttg atctaatgta
     6661 gagttgcacg tagttcttac tggcaaagaa atc
//
