LOCUS       sequence-350318-        5524 bp    DNA     circular SYN 15-DEC-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 5524)
  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 5524)
  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: pDN625
FEATURES             Location/Qualifiers
     source          1..5524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(549..1340,1341..1409))
                     /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     1172..1191
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1410..1514)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    1540..1573
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1610..1628
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1666..1688)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1788..1807
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2301..3395)
                     /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(3396..3803)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(4104..4559)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4241..4262)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4453..4472
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(4536..4916)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     4685..4707
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4699..4716
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4700..4716
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4726..4745
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4726..4744
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    4753..4860
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     4769..4785
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4770..4786
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4820..4836)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4820..4836)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4873..4893)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4873..4891)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4912..4928)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4912..4928)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4925..4947)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4936..4952
                     /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(4960..4966,4967..4984,4985..4990))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5005..5026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5143..5160)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5314..5524,1..378))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5394..5413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
       61 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      121 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
      181 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
      241 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
      301 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
      361 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
      421 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
      481 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
      541 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
      601 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
      661 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
      721 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
      781 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
      841 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
      901 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
      961 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     1021 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     1081 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     1141 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     1201 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     1261 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     1321 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     1381 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     1441 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     1501 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta taacttcgta taatgtatgc
     1561 tatacgaagt tattaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     1621 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     1681 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     1741 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     1801 gattgtactg agagtgcacc atatcgacta cgtcgttaag gccgtttctg acagagtaaa
     1861 attcttgagg gaactttcac cattatggga aatggttcaa gaaggtattg acttaaactc
     1921 catcaaatgg tcaggtcatt gagtgttttt tatttgttgt attttttttt ttttagagaa
     1981 aatcctccaa tatataaatt aggaatcata gtttcatgat tttctgttac acctaacttt
     2041 ttgtgtggtg ccctcctcct tgtcaatatt aatgttaaag tgcaattctt tttccttatc
     2101 acgttgagcc attagtatca atttgcttac ctgtattcct ttacatcctc ctttttctcc
     2161 ttcttgataa atgtatgtag attgcgtata tagtttcgtc taccctatga acatattcca
     2221 ttttgtaatt tcgtgtcgtt tctattatga atttcattta taaagtttat gtacaaatat
     2281 cataaaaaaa gagaatcttt ttaagcaagg attttcttaa cttcttcggc gacagcatca
     2341 ccgacttcgg tggtactgtt ggaaccacct aaatcaccag ttctgatacc tgcatccaaa
     2401 acctttttaa ctgcatcttc aatggcctta ccttcttcag gcaagttcaa tgacaatttc
     2461 aacatcattg cagcagacaa gatagtggcg atagggttga ccttattctt tggcaaatct
     2521 ggagcagaac cgtggcatgg ttcgtacaaa ccaaatgcgg tgttcttgtc tggcaaagag
     2581 gccaaggacg cagatggcaa caaacccaag gaacctggga taacggaggc ttcatcggag
     2641 atgatatcac caaacatgtt gctggtgatt ataataccat ttaggtgggt tgggttctta
     2701 actaggatca tggcggcaga atcaatcaat tgatgttgaa ccttcaatgt agggaattcg
     2761 ttcttgatgg tttcctccac agtttttctc cataatcttg aagaggccaa aacattagct
     2821 ttatccaagg accaaatagg caatggtggc tcatgttgta gggccatgaa agcggccatt
     2881 cttgtgattc tttgcacttc tggaacggtg tattgttcac tatcccaagc gacaccatca
     2941 ccatcgtctt cctttctctt accaaagtaa atacctccca ctaattctct gacaacaacg
     3001 aagtcagtac ctttagcaaa ttgtggcttg attggagata agtctaaaag agagtcggat
     3061 gcaaagttac atggtcttaa gttggcgtac aattgaagtt ctttacggat ttttagtaaa
     3121 ccttgttcag gtctaacact accggtaccc catttaggac cacccacagc acctaacaaa
     3181 acggcatcag ccttcttgga ggcttccagc gcctcatctg gaagtggaac acctgtagca
     3241 tcgatagcag caccaccaat taaatgattt tcgaaatcga acttgacatt ggaacgaaca
     3301 tcagaaatag ctttaagaac cttaatggct tcggctgtga tttcttgacc aacgtggtca
     3361 cctggcaaaa cgacgatctt cttaggggca gacattagaa tggtatatcc ttgaaatata
     3421 tatatatata ttgctgaaat gtaaaaggta agaaaagtta gaaagtaaga cgattgctaa
     3481 ccacctattg gaaaaaacaa taggtcctta aataatattg tcaacttcaa gtattgtgat
     3541 gcaagcattt agtcatgaac gcttctctat tctatatgaa aagccggttc cggcgctctc
     3601 acctttcctt tttctcccaa tttttcagtt gaaaaaggta tatgcgtcag gcgacctctg
     3661 aaattaacaa aaaatttcca gtcatcgaat ttgattctgt gcgatagcgc ccctgtgtgt
     3721 tctcgttatg ttgaggaaaa aaataatggt tgctaagaga ttcgaactct tgcatcttac
     3781 gatacctgag tattcccaca gttaactgcg gtcaagatat ttcttgaatc aggcgcctta
     3841 gaccgctcgg ccaaacaacc aattacttgt tgagaaatag agtataatta tcctataaat
     3901 ataacgtttt tgaacacaca tgaacaagga agtacaggac aattgatttt gaagagaatg
     3961 tggattttga tgtaattgtt gggattccat ttttaataag gcaataatat taggtatgta
     4021 gatatactag aagttctcct cgaccggtcg atatgcggtg tgaaataccg cacagatgcg
     4081 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     4141 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4201 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4261 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4321 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4381 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4441 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4501 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4561 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     4621 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     4681 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     4741 tagggcgaat tgggtaccgg gccccccctc gaggtcgacg gtatcgataa gcttgatatc
     4801 gaattcctgc agcccggggg atccactagt tctagagcgg ccgccaccgc ggtggagctc
     4861 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     4921 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4981 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5041 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5101 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5521 tagg
//
