LOCUS       sequence-104600-        6884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ADE2 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 6884)
  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 6884)
  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: pXRA2
FEATURES             Location/Qualifiers
     source          1..6884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     105..124
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        316..691
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     CDS             692..2407
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     rep_origin      complement(2612..3067)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2749..2770)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2961..2980
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(3044..3424)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3193..3215
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3207..3224
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3208..3224
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3234..3253
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3234..3252
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3261..3368
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3277..3293
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3278..3294
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3328..3344)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3328..3344)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3381..3401)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3381..3399)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3420..3436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3420..3436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3433..3455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3444..3460
                     /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(3468..3474,3475..3492,3493..3498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3513..3534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3651..3668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3822..4410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3902..3921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4581..5372,5373..5441))
                     /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     5204..5223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5442..5533)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    5568..5601
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    6176..6209
                     /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 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
      181 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      241 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      301 taagatctga attaattctt gaataataca taacttttct taaaagaatc aaagacagat
      361 aaaatttaag agatattaaa tattagtgag aagccgagaa ttttgtaaca ccaacataac
      421 actgacatct ttaacaactt ttaattatga tacatttctt acgtcatgat tgattattac
      481 agctatgctg acaaatgact cttgttgcat ggctacgaac cgggtaatac taagtgattg
      541 actcttgctg accttttatt aagaactaaa tggacaatat tatggagcat ttcatgtata
      601 aattggtgcg taaaatcgtt ggatctctct tctaagtaca tcctactata acaatcaaga
      661 aaaacaagaa aatcggacaa aacaatcaag tatggattct agaacagttg gtatattagg
      721 agggggacaa ttgggacgta tgattgttga ggcagcaaac aggctcaaca ttaagacggt
      781 aatactagat gctgaaaatt ctcctgccaa acaaataagc aactccaatg accacgttaa
      841 tggctccttt tccaatcctc ttgatatcga aaaactagct gaaaaatgtg atgtgctaac
      901 gattgagatt gagcatgttg atgttcctac actaaagaat cttcaagtaa aacatcccaa
      961 attaaaaatt tacccttctc cagaaacaat cagattgata caagacaaat atattcaaaa
     1021 agagcattta atcaaaaatg gtatagcagt tacccaaagt gttcctgtgg aacaagccag
     1081 tgagacgtcc ctattgaatg ttggaagaga tttgggtttt ccattcgtct tgaagtcgag
     1141 gactttggca tacgatggaa gaggtaactt cgttgtaaag aataaggaaa tgattccgga
     1201 agctttggaa gtactgaagg atcgtccttt gtacgccgaa aaatgggcac catttactaa
     1261 agaattagca gtcatgattg tgaggtctgt taacggttta gtgttttctt acccaattgt
     1321 agagactatc cacaaggaca atatttgtga cttatgttat gcgcctgcta gagttccgga
     1381 ctccgttcaa cttaaggcga agttgttggc agaaaatgca atcaaatctt ttcccggttg
     1441 tggtatattt ggtgtggaaa tgttctattt agaaacaggg gaattgctta ttaacgaaat
     1501 tgccccaagg cctcacaact ctggacatta taccattgat gcttgcgtca cttctcaatt
     1561 tgaagctcat ttgagatcaa tattggattt gccaatgcca aagaatttca catctttctc
     1621 caccattaca acgaacgcca ttatgctaaa tgttcttgga gacaaacata caaaagataa
     1681 agagctagaa acttgcgaaa gagcattggc gactccaggt tcctcagtgt acttatatgg
     1741 aaaagagtct agacctaaca gaaaagtagg tcacataaat attattgcct ccagtatggc
     1801 ggaatgtgaa caaaggctga actacattac aggtagaact gatattccaa tcaaaatctc
     1861 tgtcgctcaa aagttggact tggaagcaat ggtcaaacca ttggttggaa tcatcatggg
     1921 atcagactct gacttgccgg taatgtctgc cgcatgtgcg gttttaaaag attttggcgt
     1981 tccatttgaa gtgacaatag tctctgctca tagaactcca cataggatgt cagcatatgc
     2041 tatttccgca agcaagcgtg gaattaaaac aattatcgct ggagctggtg gggctgctca
     2101 cttgccaggt atggtggctg caatgacacc acttcctgtc atcggtgtgc ccgtaaaagg
     2161 ttcttgtcta gatggagtag attctttaca ttcaattgtg caaatgccta gaggtgttcc
     2221 agtagctacc gtcgctatta ataatagtac gaacgctgcg ctgttggctg tcagactgct
     2281 tggcgcttat gattcaagtt atacaacgaa aatggaacag tttttattaa agcaagaaga
     2341 agaagttctt gtcaaagcac aaaagttaga aactgtcggt tacgaagctt atctagaaaa
     2401 caagtaatat ataagtttat tgatatactt gtacagcaaa taattataaa atgatatacc
     2461 tattttttag gctttgttat gattacatca aatgtggact tcatacatag aaatcaacgc
     2521 ttacaggtgt ccttttttaa gaatttcata cataagatct atgcggtgtg aaataccgca
     2581 cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa
     2641 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     2701 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     2761 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     2821 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     2881 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     2941 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3001 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3061 ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     3121 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     3181 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
     3241 actcactata gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc
     3301 ttgatatcga attcctgcag cccgggggat ccactagttc tagagcggcc gccaccgcgg
     3361 tggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     3421 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3481 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3541 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3601 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3661 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3721 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3781 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3841 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3901 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3961 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4021 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4081 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4141 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4201 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4261 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4321 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4381 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4441 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4501 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4561 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4621 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4681 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4741 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4801 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4861 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4921 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4981 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5041 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5101 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5161 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5221 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5281 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5341 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5401 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5461 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5521 aaaaataaac aaataggggt tcagctgaag cttcgacaac ccttaatata acttcgtata
     5581 atgtatgcta tacgaagtta ttaggtctag agatccccgc gcacatttcc ccgaaaagtg
     5641 ccacctgggt ccttttcatc acgtactata aaaataatta taatttaaat tttttaatat
     5701 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
     5761 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
     5821 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
     5881 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
     5941 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
     6001 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
     6061 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
     6121 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat gcgagaaccc ttaatataac
     6181 ttcgtataat gtatgctata cgaagttatt aggtgatatc agatccacta gctataaggt
     6241 tgatattctc acgagtaagc ggcacttgct acttattgac attgcagatt tttggctaca
     6301 gaaatagtat attagagatt ataattgcta atcaaatcaa aatataaaat tagtaaacca
     6361 aaccatttat acccttcctt agtagttatg gattgttttt taatgatatt tctgcaaacc
     6421 aaagaaagat tgttatccag atagaattta gttttgatat tcattttttt gttgaagatt
     6481 gaacgccata tctgggcctc ataattcaaa agacggtgcc attatcggta gcgtttcgca
     6541 ttgtactgga tggcgcgcct tcagaaattt cacagttgat gaatcgaaaa gaatggtctc
     6601 attgcaacac gtaaggttaa gatgtccctt tttaccatta taggcaataa atgaatcata
     6661 aaacgaccgt atactggtga aatagtaggg agaacgagta cctgtagtaa aaagtataaa
     6721 tcatagttaa tcgggcaatg tccctcgatc aaggagtatt gtgtcatgtt cgagacaaac
     6781 gccaacattt ttgtttcttt tggacaaatg ttgtttgcat ttatgatccg ttatattttg
     6841 atctaatgta gagttgcacg tagttcttac tggcaaagaa atct
//
