LOCUS       sequence-350279-        6058 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast shuttle plasmid that allows conversion between low copy, high 
            copy, and integrating vectors. Low copy, LEU2-selected..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6058)
  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 6058)
  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: "G3 
            (Bethesda)."; date: "2021-12-8"; volume: "11(12). pii: 6380430. 
            doi"; pages: " 10.1093/g3journal/jkab336"
COMMENT     Sequence Label: pDN515
FEATURES             Location/Qualifiers
     source          1..6058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..41
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(79..101)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     201..220
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(714..1808)
                     /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(1809..2216)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2517..2972)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2654..2675)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2866..2885
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2949..3329)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     3098..3120
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3112..3129
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3113..3129
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3139..3158
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3139..3157
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    3166..3273
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     3182..3198
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3183..3199
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3233..3249)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3233..3249)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3286..3306)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3286..3304)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3325..3341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3325..3341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3338..3360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3349..3365
                     /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(3373..3379,3380..3397,3398..3403))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3418..3439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3556..3573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3727..4315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3807..3826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4486..5277,5278..5346))
                     /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     5109..5128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5347..5451)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5503..6006
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
       61 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
      121 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
      181 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatcga
      241 ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt caccattatg
      301 ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc attgagtgtt
      361 ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa attaggaatc
      421 atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct ccttgtcaat
      481 attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta tcaatttgct
      541 tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg tagattgcgt
      601 atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc gtttctatta
      661 tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc tttttaagca
      721 aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact gttggaacca
      781 cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc ttcaatggcc
      841 ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga caagatagtg
      901 gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca tggttcgtac
      961 aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg caacaaaccc
     1021 aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat gttgctggtg
     1081 attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc agaatcaatc
     1141 aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc cacagttttt
     1201 ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat aggcaatggt
     1261 ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac ttctggaacg
     1321 gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct cttaccaaag
     1381 taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc aaattgtggc
     1441 ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct taagttggcg
     1501 tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac actaccggta
     1561 ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt ggaggcttcc
     1621 agcgcctcat ctggaagtgg aacacctgta gcatcgatag cagcaccacc aattaaatga
     1681 ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag aaccttaatg
     1741 gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat cttcttaggg
     1801 gcagacatta gaatggtata tccttgaaat atatatatat atattgctga aatgtaaaag
     1861 gtaagaaaag ttagaaagta agacgattgc taaccaccta ttggaaaaaa caataggtcc
     1921 ttaaataata ttgtcaactt caagtattgt gatgcaagca tttagtcatg aacgcttctc
     1981 tattctatat gaaaagccgg ttccggcgct ctcacctttc ctttttctcc caatttttca
     2041 gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa caaaaaattt ccagtcatcg
     2101 aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt atgttgagga aaaaaataat
     2161 ggttgctaag agattcgaac tcttgcatct tacgatacct gagtattccc acagttaact
     2221 gcggtcaaga tatttcttga atcaggcgcc ttagaccgct cggccaaaca accaattact
     2281 tgttgagaaa tagagtataa ttatcctata aatataacgt ttttgaacac acatgaacaa
     2341 ggaagtacag gacaattgat tttgaagaga atgtggattt tgatgtaatt gttgggattc
     2401 catttttaat aaggcaataa tattaggtat gtagatatac tagaagttct cctcgaccgg
     2461 tcgatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     2521 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2581 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2641 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2701 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2761 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2821 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2881 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2941 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     3001 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     3061 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     3121 acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc
     3181 ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact
     3241 agttctagag cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     3301 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3361 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3421 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3481 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3541 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3601 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3661 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3721 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3781 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3841 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3901 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3961 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4021 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4081 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4141 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4201 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4261 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4321 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4381 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4441 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4501 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4561 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4621 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4681 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4741 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4801 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4861 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4921 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4981 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5041 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5101 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5161 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5221 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5281 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5341 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5401 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5461 ccgaaaagtg ccacctgacg tcctaggcat gcggtccttt tcatcacgtg ctataaaaat
     5521 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     5581 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     5641 acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca
     5701 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     5761 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     5821 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     5881 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     5941 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     6001 gcgatccgtc taagaaacca ttattatcgc atgcctagga cgtctaagaa accattat
//
