LOCUS       sequence-350295-        4935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast shuttle plasmid that allows conversion between low copy, high 
            copy, and integrating vectors. Low copy, URA3-selected..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4935)
  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 4935)
  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: pDN516
FEATURES             Location/Qualifiers
     source          1..4935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     266..284
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(322..344)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     444..463
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        487..702
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             703..1506
                     /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(1637..2092)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1774..1795)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1986..2005
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2069..2449)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2218..2240
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2232..2249
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2233..2249
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2259..2278
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2259..2277
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2286..2393
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2302..2318
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2303..2319
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2353..2369)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2353..2369)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2406..2426)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2406..2424)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2445..2461)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2445..2461)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2458..2480)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2469..2485
                     /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(2493..2499,2500..2517,2518..2523))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2538..2559
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2676..2693)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2847..3435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2927..2946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3606..4397,4398..4466))
                     /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     4229..4248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4467..4571)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(4623..4935,1..191)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 agtacgcttt ttgttgaaat tttttaaacc tttgtttatt tttttttctt cattccgtaa
       61 ctcttctacc ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa
      121 cacagagtaa attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttaca
      181 ggcaagcgat ccgtctaaga aaccattatt atcgcatgcc taggacgtct aagaaaccat
      241 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
      301 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
      361 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      421 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatac
      481 caccttttca attcatcatt ttttttttat tctttttttt gatttcggtt tccttgaaat
      541 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
      601 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
      661 caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa tcatgtcgaa agctacatat
      721 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
      781 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
      841 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
      901 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
      961 ttactcttcg aagacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     1021 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     1081 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     1141 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     1201 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     1261 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     1321 gatgacaagg gagacgcatt gggtcaacag tatagaaccg tggatgatgt ggtctctaca
     1381 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     1441 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     1501 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     1561 atttaattat atcagttatt accctatgcg gtgtgaaata ccgcacagat gcgtaaggag
     1621 aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     1681 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     1741 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     1801 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     1861 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     1921 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     1981 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2041 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc
     2101 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2161 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2221 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
     2281 aattgggtac cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattcc
     2341 tgcagcccgg gggatccact agttctagag cggccgccac cgcggtggag ctccagcttt
     2401 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     2461 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2521 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2581 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2641 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2701 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2761 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2821 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2881 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2941 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3001 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3061 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3121 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3181 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3241 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3301 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3361 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3421 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3481 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3541 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3601 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3661 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3721 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3781 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3841 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3901 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3961 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4021 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4081 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4141 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4201 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4261 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4321 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4381 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4441 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4501 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4561 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcctaggcat gcggtccttt
     4621 tcatcacgtg ctataaaaat aattataatt taaatttttt aatataaata tataaattaa
     4681 aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg aagatgtaaa
     4741 agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct gctctcaggt
     4801 attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa atcctgtgat
     4861 tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt tcttgtctaa
     4921 taaatatata tgtaa
//
