LOCUS       sequence-350312-        6108 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 6108)
  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 6108)
  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: pDN615
FEATURES             Location/Qualifiers
     source          1..6108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(591..1382,1383..1451))
                     /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     1214..1233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1452..1556)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    1582..1615
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1651..2154
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     protein_bind    2166..2199
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2236..2254
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2292..2314)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2414..2433
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2927..4021)
                     /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(4022..4429)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(4730..5185)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4867..4888)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5079..5098
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(5162..5542)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     5311..5333
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5325..5342
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5326..5342
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5352..5371
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5352..5370
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5379..5486
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     5395..5411
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5396..5412
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5446..5462)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5446..5462)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5499..5519)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5499..5517)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5538..5554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5538..5554)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5551..5573)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5562..5578
                     /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(5586..5592,5593..5610,5611..5616))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5631..5652
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5769..5786)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5940..6108,1..420))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6020..6039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
       61 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      361 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      421 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      481 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      541 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      601 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      661 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      721 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      781 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      841 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      901 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      961 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1021 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1081 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1141 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1201 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1261 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1321 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1381 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1441 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1501 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1561 tttccccgaa aagtgccacc tataacttcg tataatgtat gctatacgaa gttattgacg
     1621 tccccgaaaa gtgccacctg ggtccttttc atcacgtgct ataaaaataa ttataattta
     1681 aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa
     1741 aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc
     1801 ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt
     1861 agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat
     1921 atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga
     1981 aattttttaa acctttgttt attttttttt cttcattccg taactcttct accttcttta
     2041 tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca
     2101 aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtccg
     2161 acgtcataac ttcgtataat gtatgctata cgaagttatt aagaaaccat tattatcatg
     2221 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     2281 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     2341 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     2401 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat cgactacgtc
     2461 gttaaggccg tttctgacag agtaaaattc ttgagggaac tttcaccatt atgggaaatg
     2521 gttcaagaag gtattgactt aaactccatc aaatggtcag gtcattgagt gttttttatt
     2581 tgttgtattt tttttttttt agagaaaatc ctccaatata taaattagga atcatagttt
     2641 catgattttc tgttacacct aactttttgt gtggtgccct cctccttgtc aatattaatg
     2701 ttaaagtgca attctttttc cttatcacgt tgagccatta gtatcaattt gcttacctgt
     2761 attcctttac atcctccttt ttctccttct tgataaatgt atgtagattg cgtatatagt
     2821 ttcgtctacc ctatgaacat attccatttt gtaatttcgt gtcgtttcta ttatgaattt
     2881 catttataaa gtttatgtac aaatatcata aaaaaagaga atctttttaa gcaaggattt
     2941 tcttaacttc ttcggcgaca gcatcaccga cttcggtggt actgttggaa ccacctaaat
     3001 caccagttct gatacctgca tccaaaacct ttttaactgc atcttcaatg gccttacctt
     3061 cttcaggcaa gttcaatgac aatttcaaca tcattgcagc agacaagata gtggcgatag
     3121 ggttgacctt attctttggc aaatctggag cagaaccgtg gcatggttcg tacaaaccaa
     3181 atgcggtgtt cttgtctggc aaagaggcca aggacgcaga tggcaacaaa cccaaggaac
     3241 ctgggataac ggaggcttca tcggagatga tatcaccaaa catgttgctg gtgattataa
     3301 taccatttag gtgggttggg ttcttaacta ggatcatggc ggcagaatca atcaattgat
     3361 gttgaacctt caatgtaggg aattcgttct tgatggtttc ctccacagtt tttctccata
     3421 atcttgaaga ggccaaaaca ttagctttat ccaaggacca aataggcaat ggtggctcat
     3481 gttgtagggc catgaaagcg gccattcttg tgattctttg cacttctgga acggtgtatt
     3541 gttcactatc ccaagcgaca ccatcaccat cgtcttcctt tctcttacca aagtaaatac
     3601 ctcccactaa ttctctgaca acaacgaagt cagtaccttt agcaaattgt ggcttgattg
     3661 gagataagtc taaaagagag tcggatgcaa agttacatgg tcttaagttg gcgtacaatt
     3721 gaagttcttt acggattttt agtaaacctt gttcaggtct aacactaccg gtaccccatt
     3781 taggaccacc cacagcacct aacaaaacgg catcagcctt cttggaggct tccagcgcct
     3841 catctggaag tggaacacct gtagcatcga tagcagcacc accaattaaa tgattttcga
     3901 aatcgaactt gacattggaa cgaacatcag aaatagcttt aagaacctta atggcttcgg
     3961 ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac gatcttctta ggggcagaca
     4021 ttagaatggt atatccttga aatatatata tatatattgc tgaaatgtaa aaggtaagaa
     4081 aagttagaaa gtaagacgat tgctaaccac ctattggaaa aaacaatagg tccttaaata
     4141 atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc atgaacgctt ctctattcta
     4201 tatgaaaagc cggttccggc gctctcacct ttcctttttc tcccaatttt tcagttgaaa
     4261 aaggtatatg cgtcaggcga cctctgaaat taacaaaaaa tttccagtca tcgaatttga
     4321 ttctgtgcga tagcgcccct gtgtgttctc gttatgttga ggaaaaaaat aatggttgct
     4381 aagagattcg aactcttgca tcttacgata cctgagtatt cccacagtta actgcggtca
     4441 agatatttct tgaatcaggc gccttagacc gctcggccaa acaaccaatt acttgttgag
     4501 aaatagagta taattatcct ataaatataa cgtttttgaa cacacatgaa caaggaagta
     4561 caggacaatt gattttgaag agaatgtgga ttttgatgta attgttggga ttccattttt
     4621 aataaggcaa taatattagg tatgtagata tactagaagt tctcctcgac cggtcgatat
     4681 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     4741 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4801 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4861 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4921 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4981 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5041 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5101 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5161 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     5221 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     5281 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     5341 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg
     5401 tcgacggtat cgataagctt gatatcgaat tcctgcagcc cgggggatcc actagttcta
     5461 gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg
     5521 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5581 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5641 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5701 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5761 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5821 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5881 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5941 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6001 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6061 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctc
//
