LOCUS       sequence-350296-        5825 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast shuttle plasmid that allows conversion between low copy, high 
            copy, and integrating vectors. High copy, HIS3-selected..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5825)
  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 5825)
  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: pDN523
FEATURES             Location/Qualifiers
     source          1..5825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     52..70
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(108..130)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     230..249
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        396..583
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             584..1246
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1506..1961)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1643..1664)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1855..1874
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1938..2318)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2087..2109
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2101..2118
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2102..2118
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2128..2147
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2128..2146
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2155..2262
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2171..2187
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2172..2188
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2222..2238)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2222..2238)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2275..2295)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2275..2293)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2314..2330)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2314..2330)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2327..2349)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2338..2354
                     /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(2362..2368,2369..2386,2387..2392))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2407..2428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2545..2562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2716..3304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2796..2815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3475..4266,4267..4335))
                     /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     4098..4117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4336..4440)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4482..5824
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5445..5492)
                     /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)."
ORIGIN
        1 catgcctagg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
       61 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
      121 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
      181 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
      241 cagattgtac tgagagtgca ccataaattc ccgttttaag agcttggtga gcgctaggag
      301 tcactgccag gtatcgtttg aacacggcat tagtcaggga agtcataaca cagtcctttc
      361 ccgcaatttt ctttttctat tactcttggc ctcctctagt acactctata tttttttatg
      421 cctcggtaat gattttcatt tttttttttc cacctagcgg atgactcttt ttttttctta
      481 gcgattggca ttatcacata atgaattata cattatataa agtaatgtga tttcttcgaa
      541 gaatatacta aaaaatgagc aggcaagata aacgaaggca aagatgacag agcagaaagc
      601 cctagtaaag cgtattacaa atgaaaccaa gattcagatt gcgatctctt taaagggtgg
      661 tcccctagcg atagagcact cgatcttccc agaaaaagag gcagaagcag tagcagaaca
      721 ggccacacaa tcgcaagtga ttaacgtcca cacaggtata gggtttctgg accatatgat
      781 acatgctctg gccaagcatt ccggctggtc gctaatcgtt gagtgcattg gtgacttaca
      841 catagacgac catcacacca ctgaagactg cgggattgct ctcggtcaag cttttaaaga
      901 ggccctaggg gccgtgcgtg gagtaaaaag gtttggatca ggatttgcgc ctttggatga
      961 ggcactttcc agagcggtgg tagatctttc gaacaggccg tacgcagttg tcgaacttgg
     1021 tttgcaaagg gagaaagtag gagatctctc ttgcgagatg atcccgcatt ttcttgaaag
     1081 ctttgcagag gctagcagaa ttaccctcca cgttgattgt ctgcgaggca agaatgatca
     1141 tcaccgtagt gagagtgcgt tcaaggctct tgcggttgcc ataagagaag ccacctcgcc
     1201 caatggtacc aacgatgttc cctccaccaa aggtgttctt atgtagtgac accgattatt
     1261 taaagctgca gcatacgata tatatacatg tgtatatatg tatacctatg aatgtcagta
     1321 agtatgtata cgaacagtat gatactgaag atgacaaggt aatgcatcat tctatacgtg
     1381 tcattctgaa cgaggcgcgc tttccttttt tctttttgct ttttcttttt ttttctcttg
     1441 aactcgacgg atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     1501 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     1561 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     1621 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     1681 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     1741 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     1801 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     1861 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     1921 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     1981 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     2041 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     2101 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
     2161 gggccccccc tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg
     2221 ggatccacta gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta
     2281 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2341 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2401 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2461 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2521 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3121 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4381 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4441 cacatttccc cgaaaagtgc cacctgacgt cctaggcatg caacgaagca tctgtgcttc
     4501 attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     4561 gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg
     4621 cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     4681 gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat
     4741 ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc
     4801 atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     4861 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     4921 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     4981 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     5041 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     5101 tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat
     5161 tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa
     5221 cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta
     5281 ggttatatag ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg
     5341 tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt
     5401 tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt
     5461 ctagagaata ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg agcgcttccg
     5521 aaaatgcaac gcgagctgcg cacatacagc tcactgttca cgtcgcacct atatctgcgt
     5581 gttgcctgta tatatatata catgagaaga acggcatagt gcgtgtttat gcttaaatgc
     5641 gtacttatat gcgtctattt atgtaggatg aaaggtagtc tagtacctcc tgtgatatta
     5701 tcccattcca tgcggggtat cgtatgcttc cttcagcact accctttagc tgttctatat
     5761 gctgccactc ctcaattgga ttagtctcat ccttcaatgc tatcatttcc tttgatattg
     5821 gatcg
//
