LOCUS       sequence-350319-        5792 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 5792)
  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 5792)
  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: pDN626
FEATURES             Location/Qualifiers
     source          1..5792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(320..367)
                     /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)."
     protein_bind    706..739
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     777..795
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(833..855)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     955..974
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        998..1213
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1214..2017
                     /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(2148..2603)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2285..2306)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2497..2516
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2580..2960)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2729..2751
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2743..2760
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2744..2760
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2770..2789
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2770..2788
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2797..2904
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2813..2829
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2814..2830
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2864..2880)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2864..2880)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2917..2937)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2917..2935)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2956..2972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2956..2972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2969..2991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2980..2996
                     /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(3004..3010,3011..3028,3029..3034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3049..3070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3187..3204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3358..3946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3438..3457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4117..4908,4909..4977))
                     /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     4740..4759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4978..5082)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    5108..5141
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      join(5149..5792,1..699)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
       61 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
      121 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
      181 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
      241 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt
      301 tggttttcaa aagcgctctg aagttcctat actttctaga gaataggaac ttcggaatag
      361 gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat
      421 acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat atatacatga
      481 gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc tatttatgta
      541 ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg ggtatcgtat
      601 gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa ttggattagt
      661 ctcatccttc aatgctatca tttcctttga tattggatcg acgtcataac ttcgtataat
      721 gtatgctata cgaagttatt taagaaacca ttattatcat gacattaacc tataaaaata
      781 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
      841 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
      901 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
      961 cagagcagat tgtactgaga gtgcaccata ccaccttttc aattcatcat ttttttttta
     1021 ttcttttttt tgatttcggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa
     1081 ggaagaacga aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga
     1141 agaaacatga aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa
     1201 acgaagataa atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc
     1261 tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt
     1321 ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg
     1381 tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc
     1441 gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa aatttgctga
     1501 cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc
     1561 agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc
     1621 ggcagaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa
     1681 gggctcccta tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa
     1741 agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga
     1801 ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat tgggtcaaca
     1861 gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg ttggaagagg
     1921 actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg
     1981 ggaagcatat ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc
     2041 atgtatacta aactcacaaa ttagagcttc aatttaatta tatcagttat taccctatgc
     2101 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt
     2161 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2221 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2281 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2341 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2401 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2461 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2521 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2581 taatgcgccg ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg
     2641 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     2701 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     2761 gagcgcgcgt aatacgactc actatagggc gaattgggta ccgggccccc cctcgaggtc
     2821 gacggtatcg ataagcttga tatcgaattc ctgcagcccg ggggatccac tagttctaga
     2881 gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg
     2941 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3001 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3061 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3121 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3181 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3241 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3301 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3361 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3421 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3481 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3541 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3601 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3661 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3721 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3781 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3841 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3901 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3961 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4021 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4081 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4141 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4201 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4261 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4321 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4381 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4441 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4501 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4561 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4621 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4681 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4741 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4801 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4861 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4921 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4981 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5041 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5101 gccacctata acttcgtata atgtatgcta tacgaagtta ttgacgtcaa cgaagcatct
     5161 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     5221 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     5281 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     5341 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     5401 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     5461 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     5521 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     5581 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     5641 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     5701 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     5761 ggtttcttct attttgtctc tatatactac gt
//
