LOCUS       sequence-350298-        6865 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast shuttle plasmid that allows conversion between low copy, high 
            copy, and integrating vectors. High copy, LEU2-selected..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6865)
  AUTHORS   Nickerson DP, Quinn MA, Milnes JM
  TITLE     Rapid conversion of replicating and integrating Saccharomyces 
            cerevisiae plasmid vectors via Cre recombinase.
  JOURNAL   G3 (Bethesda). 2021 Dec 8;11(12). pii: 6380430. doi: 
            10.1093/g3journal/jkab336.
  PUBMED    34599813
REFERENCE   2  (bases 1 to 6865)
  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: "G3 
            (Bethesda)."; date: "2021-12-8"; volume: "11(12). pii: 6380430. 
            doi"; pages: " 10.1093/g3journal/jkab336"
COMMENT     Sequence Label: pDN525
FEATURES             Location/Qualifiers
     source          1..6865
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     83..101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(139..161)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     261..280
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(774..1868)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1869..2276)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2577..3032)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2714..2735)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2926..2945
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(3009..3389)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3158..3180
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3172..3189
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3173..3189
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3199..3218
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3199..3217
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3226..3333
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3242..3258
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3243..3259
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3293..3309)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3293..3309)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3346..3366)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3346..3364)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3385..3401)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3385..3401)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3398..3420)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3409..3425
                     /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(3433..3439,3440..3457,3458..3463))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3478..3499
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3616..3633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3787..4375)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3867..3886)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4546..5337,5338..5406))
                     /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     5169..5188
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5407..5511)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5553..6865,1..30)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(6516..6563)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 caatgctatc atttcctttg atattggatc gcatgcctag gacgtctaag aaaccattat
       61 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
      121 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
      181 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
      241 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatcga
      301 ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt caccattatg
      361 ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc attgagtgtt
      421 ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa attaggaatc
      481 atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct ccttgtcaat
      541 attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta tcaatttgct
      601 tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg tagattgcgt
      661 atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc gtttctatta
      721 tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc tttttaagca
      781 aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact gttggaacca
      841 cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc ttcaatggcc
      901 ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga caagatagtg
      961 gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca tggttcgtac
     1021 aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg caacaaaccc
     1081 aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat gttgctggtg
     1141 attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc agaatcaatc
     1201 aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc cacagttttt
     1261 ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat aggcaatggt
     1321 ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac ttctggaacg
     1381 gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct cttaccaaag
     1441 taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc aaattgtggc
     1501 ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct taagttggcg
     1561 tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac actaccggta
     1621 ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt ggaggcttcc
     1681 agcgcctcat ctggaagtgg aacacctgta gcatcgatag cagcaccacc aattaaatga
     1741 ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag aaccttaatg
     1801 gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat cttcttaggg
     1861 gcagacatta gaatggtata tccttgaaat atatatatat atattgctga aatgtaaaag
     1921 gtaagaaaag ttagaaagta agacgattgc taaccaccta ttggaaaaaa caataggtcc
     1981 ttaaataata ttgtcaactt caagtattgt gatgcaagca tttagtcatg aacgcttctc
     2041 tattctatat gaaaagccgg ttccggcgct ctcacctttc ctttttctcc caatttttca
     2101 gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa caaaaaattt ccagtcatcg
     2161 aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt atgttgagga aaaaaataat
     2221 ggttgctaag agattcgaac tcttgcatct tacgatacct gagtattccc acagttaact
     2281 gcggtcaaga tatttcttga atcaggcgcc ttagaccgct cggccaaaca accaattact
     2341 tgttgagaaa tagagtataa ttatcctata aatataacgt ttttgaacac acatgaacaa
     2401 ggaagtacag gacaattgat tttgaagaga atgtggattt tgatgtaatt gttgggattc
     2461 catttttaat aaggcaataa tattaggtat gtagatatac tagaagttct cctcgaccgg
     2521 tcgatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     2581 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2641 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2701 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2761 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2821 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2881 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2941 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3001 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     3061 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     3121 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     3181 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
     3241 ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact
     3301 agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     3361 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3421 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3481 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3541 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3601 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3661 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3721 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3781 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3841 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3901 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3961 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4021 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4081 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4141 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4201 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4261 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4321 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4381 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4441 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4501 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4561 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4621 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4681 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4741 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4801 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4861 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4921 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4981 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5041 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5101 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5161 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5221 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5281 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5341 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5401 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5461 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5521 ccgaaaagtg ccacctgacg tcctaggcat gcaacgaagc atctgtgctt cattttgtag
     5581 aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta
     5641 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     5701 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     5761 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     5821 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     5881 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     5941 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     6001 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     6061 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     6121 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     6181 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     6241 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     6301 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     6361 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     6421 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     6481 gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagagaat
     6541 aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc gaaaatgcaa
     6601 cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg tgttgcctgt
     6661 atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg cgtacttata
     6721 tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt atcccattcc
     6781 atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata tgctgccact
     6841 cctcaattgg attagtctca tcctt
//
