LOCUS       sequence-350299-        5742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast shuttle plasmid that allows conversion between low copy, high 
            copy, and integrating vectors. High copy, URA3-selected..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5742)
  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 5742)
  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: pDN526
FEATURES             Location/Qualifiers
     source          1..5742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..70)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     170..189
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        213..428
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             429..1232
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1363..1818)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1500..1521)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1712..1731
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1795..2175)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1944..1966
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1958..1975
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1959..1975
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1985..2004
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1985..2003
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2012..2119
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2028..2044
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2029..2045
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2079..2095)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2079..2095)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2132..2152)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2132..2150)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2171..2187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2171..2187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2184..2206)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2195..2211
                     /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(2219..2225,2226..2243,2244..2249))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2264..2285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2402..2419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2573..3161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2653..2672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3332..4123,4124..4192))
                     /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     3955..3974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4193..4297)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4339..5681
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5302..5349)
                     /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)."
     primer_bind     join(5734..5742,1..10)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
       61 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
      121 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
      181 cagattgtac tgagagtgca ccataccacc ttttcaattc atcatttttt ttttattctt
      241 ttttttgatt tcggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag
      301 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa
      361 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa
      421 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
      481 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
      541 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
      601 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
      661 aggcattatc cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg
      721 gtaatacagt caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca
      781 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
      841 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
      901 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
      961 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
     1021 tgattatgac acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata
     1081 gaaccgtgga tgatgtggtc tctacaggat ctgacattat tattgttgga agaggactat
     1141 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     1201 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta
     1261 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc tatgcggtgt
     1321 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata
     1381 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1441 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1501 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1561 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1621 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1681 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1741 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1801 cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     1861 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     1921 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg
     1981 cgcgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcgacgg
     2041 tatcgataag cttgatatcg aattcctgca gcccggggga tccactagtt ctagagcggc
     2101 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     2161 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2221 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2281 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2341 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2401 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2461 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2521 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2581 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2641 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2701 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2761 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2821 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2881 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2941 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3001 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3061 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3121 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3181 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3241 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3301 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3361 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3421 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3481 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3541 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3601 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3661 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3721 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3781 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3841 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3901 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3961 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4021 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4081 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4141 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4201 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4261 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4321 ctgacgtcct aggcatgcaa cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac
     4381 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     4441 aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat
     4501 gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga
     4561 aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt ttgttctaca
     4621 aaaatgcatc ccgagagcgc tatttttcta acaaagcatc ttagattact ttttttctcc
     4681 tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc cgttaaggtt
     4741 agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga ctccacttcc
     4801 cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa ggcatccccg
     4861 attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg atagcgttga
     4921 tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc tatatactac
     4981 gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat agttcttact
     5041 acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta gaggtcgagt
     5101 ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga tatagcacag
     5161 agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt cgcaatattt
     5221 tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtct tcagagcgct
     5281 tttggttttc aaaagcgctc tgaagttcct atactttcta gagaatagga acttcggaat
     5341 aggaacttca aagcgtttcc gaaaacgagc gcttccgaaa atgcaacgcg agctgcgcac
     5401 atacagctca ctgttcacgt cgcacctata tctgcgtgtt gcctgtatat atatatacat
     5461 gagaagaacg gcatagtgcg tgtttatgct taaatgcgta cttatatgcg tctatttatg
     5521 taggatgaaa ggtagtctag tacctcctgt gatattatcc cattccatgc ggggtatcgt
     5581 atgcttcctt cagcactacc ctttagctgt tctatatgct gccactcctc aattggatta
     5641 gtctcatcct tcaatgctat catttccttt gatattggat cgcatgccta ggacgtctaa
     5701 gaaaccatta ttatcatgac attaacctat aaaaataggc gt
//
