LOCUS       sequence-104606-        6080 bp    DNA     circular SYN 15-APR-2025
DEFINITION  HPH 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 6080)
  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 6080)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pXRhMX
FEATURES             Location/Qualifiers
     source          1..6080
                     /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..1716
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        141..484
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             485..1513
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     CDS             485..1513
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1519..1716
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     rep_origin      complement(1808..2263)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1945..1966)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2157..2176
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2240..2620)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2389..2411
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2403..2420
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2404..2420
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2430..2449
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2430..2448
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2457..2564
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2473..2489
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2474..2490
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2524..2540)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2524..2540)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2577..2597)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2577..2595)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2616..2632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2616..2632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2629..2651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2640..2656
                     /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(2664..2670,2671..2688,2689..2694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2709..2730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2847..2864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3018..3606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3098..3117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3777..4568,4569..4637))
                     /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     4400..4419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4638..4729)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    4764..4797
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5372..5405
                     /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 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
      541 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
      601 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
      661 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
      721 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
      781 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
      841 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
      901 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
      961 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     1021 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     1081 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     1141 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     1201 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     1261 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     1321 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     1381 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     1441 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     1501 ggcaaaggaa taatcagtac tgacaataaa aagattcttg ttttcaagaa cttgtcattt
     1561 gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt gatttatatt
     1621 ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa agtaatatca
     1681 tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata ctaacgccgc
     1741 catccagtgt cgaaaactgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     1801 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     1861 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     1921 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     1981 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     2041 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     2101 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     2161 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     2221 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     2281 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     2341 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     2401 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
     2461 ccgggccccc cctcgaggtc gacggtatcg ataagcttga tatcgaattc ctgcagcccg
     2521 ggggatccac tagttctaga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     2581 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     2641 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     2701 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     2761 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     2821 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     2881 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     2941 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3001 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3061 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3121 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3181 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3241 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3301 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3361 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3421 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     3481 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3541 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     3601 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     3661 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3721 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3781 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3841 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3901 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3961 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4021 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4081 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4141 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4201 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4261 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4321 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4381 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4441 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4501 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4561 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     4621 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     4681 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttcag
     4741 ctgaagcttc gacaaccctt aatataactt cgtataatgt atgctatacg aagttattag
     4801 gtctagagat ccccgcgcac atttccccga aaagtgccac ctgggtcctt ttcatcacgt
     4861 actataaaaa taattataat ttaaattttt taatataaat atataaatta aaaatagaaa
     4921 gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag
     4981 ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc
     5041 gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga ttttacattt
     5101 tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta ataaatatat
     5161 atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt tttcttcatt
     5221 ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa cataaaaata
     5281 aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa
     5341 gttacaggca agcgatgcga gaacccttaa tataacttcg tataatgtat gctatacgaa
     5401 gttattaggt gatatcagat ccactagcta taaggttgat attctcacga gtaagcggca
     5461 cttgctactt attgacattg cagatttttg gctacagaaa tagtatatta gagattataa
     5521 ttgctaatca aatcaaaata taaaattagt aaaccaaacc atttataccc ttccttagta
     5581 gttatggatt gttttttaat gatatttctg caaaccaaag aaagattgtt atccagatag
     5641 aatttagttt tgatattcat ttttttgttg aagattgaac gccatatctg ggcctcataa
     5701 ttcaaaagac ggtgccatta tcggtagcgt ttcgcattgt actggatggc gcgccttcag
     5761 aaatttcaca gttgatgaat cgaaaagaat ggtctcattg caacacgtaa ggttaagatg
     5821 tcccttttta ccattatagg caataaatga atcataaaac gaccgtatac tggtgaaata
     5881 gtagggagaa cgagtacctg tagtaaaaag tataaatcat agttaatcgg gcaatgtccc
     5941 tcgatcaagg agtattgtgt catgttcgag acaaacgcca acatttttgt ttcttttgga
     6001 caaatgttgt ttgcatttat gatccgttat attttgatct aatgtagagt tgcacgtagt
     6061 tcttactggc aaagaaatct
//
