LOCUS       sequence-104614-        5624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nat1 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 5624)
  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 5624)
  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: pXRnMX
FEATURES             Location/Qualifiers
     source          1..5624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     gene            142..1261
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        142..485
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             486..1058
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDTEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1064..1261
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(1353..1808)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1490..1511)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1702..1721
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1785..2165)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1934..1956
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1948..1965
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1949..1965
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1975..1994
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1975..1993
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2002..2109
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2018..2034
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2019..2035
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2069..2085)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2069..2085)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2122..2142)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2122..2140)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2161..2177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2161..2177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2174..2196)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2185..2201
                     /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(2209..2215,2216..2233,2234..2239))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2254..2275
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2392..2409)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2563..3151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2643..2662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3322..4113,4114..4182))
                     /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     3945..3964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4183..4274)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    4309..4342
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4917..4950
                     /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 agacatggag gcccagaata ccctccttga cagtcttgac
      181 gtgcgcagct caggggcatg atgtgactgt cgcccgtaca tttagcccat acatccccat
      241 gtataatcat ttgcatccat acattttgat ggccgcacgg cgcgaagcaa aaattacggc
      301 tcctcgctgc agacctgcga gcagggaaac gctcccctca cagacgcgtt gaattgtccc
      361 cacgccgcgc ccctgtagag aaatataaaa ggttaggatt tgccactgag gttcttcttt
      421 catatacttc cttttaaaat cttgctagga tacagttctc acatcacatc cgaacataaa
      481 caaccatggg taccactctt gacgacacgg cttaccggta ccgcaccagt gtcccggggg
      541 acaccgaggc catcgaggca ctggatgggt ccttcaccac cgacaccgtc ttccgcgtca
      601 ccgccaccgg ggacggcttc accctgcggg aggtgccggt ggacccgccc ctgaccaagg
      661 tgttccccga cgacgaatcg gacgacgaat cggacgacgg ggaggacggc gacccggact
      721 cccggacgtt cgtcgcgtac ggggacgacg gcgacctggc gggcttcgtg gtcgtctcgt
      781 actccggctg gaaccgccgg ctgaccgtcg aggacatcga ggtcgccccg gagcaccggg
      841 ggcacggggt cgggcgcgcg ttgatggggc tcgcgacgga gttcgcccgc gagcggggcg
      901 ccgggcacct ctggctggag gtcaccaacg tcaacgcacc ggcgatccac gcgtaccggc
      961 ggatggggtt caccctctgc ggcctggaca ccgccctgta cgacggcacc gcctcggacg
     1021 gcgagcaggc gctctacatg agcatgccct gcccctaatc agtactgaca ataaaaagat
     1081 tcttgttttc aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta
     1141 atcaaatgtt agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa
     1201 gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact
     1261 gctgtcgatt cgatactaac gccgccatcc agtgtcgaaa actgcggtgt gaaataccgc
     1321 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     1381 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     1441 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     1501 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     1561 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     1621 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     1681 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     1741 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     1801 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     1861 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     1921 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
     1981 gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcgacgg tatcgataag
     2041 cttgatatcg aattcctgca gcccggggga tccactagtt ctagagcggc cgccaccgcg
     2101 gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     2161 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2221 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2281 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2341 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2401 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2461 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2521 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2581 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2641 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2701 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2761 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2821 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2881 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2941 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3001 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3061 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3121 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3181 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3241 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3301 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3361 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3421 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3481 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3541 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3601 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3661 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3721 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3781 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3841 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3901 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3961 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4021 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4081 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4141 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4201 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4261 gaaaaataaa caaatagggg ttcagctgaa gcttcgacaa cccttaatat aacttcgtat
     4321 aatgtatgct atacgaagtt attaggtcta gagatccccg cgcacatttc cccgaaaagt
     4381 gccacctggg tccttttcat cacgtactat aaaaataatt ataatttaaa ttttttaata
     4441 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     4501 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     4561 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     4621 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     4681 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     4741 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     4801 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     4861 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tgcgagaacc cttaatataa
     4921 cttcgtataa tgtatgctat acgaagttat taggtgatat cagatccact agctataagg
     4981 ttgatattct cacgagtaag cggcacttgc tacttattga cattgcagat ttttggctac
     5041 agaaatagta tattagagat tataattgct aatcaaatca aaatataaaa ttagtaaacc
     5101 aaaccattta tacccttcct tagtagttat ggattgtttt ttaatgatat ttctgcaaac
     5161 caaagaaaga ttgttatcca gatagaattt agttttgata ttcatttttt tgttgaagat
     5221 tgaacgccat atctgggcct cataattcaa aagacggtgc cattatcggt agcgtttcgc
     5281 attgtactgg atggcgcgcc ttcagaaatt tcacagttga tgaatcgaaa agaatggtct
     5341 cattgcaaca cgtaaggtta agatgtccct ttttaccatt ataggcaata aatgaatcat
     5401 aaaacgaccg tatactggtg aaatagtagg gagaacgagt acctgtagta aaaagtataa
     5461 atcatagtta atcgggcaat gtccctcgat caaggagtat tgtgtcatgt tcgagacaaa
     5521 cgccaacatt tttgtttctt ttggacaaat gttgtttgca tttatgatcc gttatatttt
     5581 gatctaatgt agagttgcac gtagttctta ctggcaaaga aatc
//
