LOCUS       sequence-350297-        5648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast shuttle plasmid that allows conversion between low copy, high 
            copy, and integrating vectors. High copy, TRP1-selected..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5648)
  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 5648)
  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: pDN524
FEATURES             Location/Qualifiers
     source          1..5648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..67)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     167..186
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        207..488
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             489..1163
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1266..1721)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1403..1424)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1615..1634
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1698..2078)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1847..1869
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1861..1878
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1862..1878
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1888..1907
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1888..1906
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1915..2022
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     1931..1947
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1932..1948
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1982..1998)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1982..1998)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2035..2055)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2035..2053)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2074..2090)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2074..2090)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2087..2109)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2098..2114
                     /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(2122..2128,2129..2146,2147..2152))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2167..2188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2305..2322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2476..3064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2556..2575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3235..4026,4027..4095))
                     /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     3858..3877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4096..4200)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4242..5584
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5205..5252)
                     /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(5637..5648,1..7)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
       61 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
      121 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
      181 attgtactga gagtgcacca tagatcaacg acattactat atatataata taggaagcat
      241 ttaatagaac agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag
      301 tggaagatat tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc
      361 ttttcttttt ttgccgatta agaattaatt cggtcgaaaa aagaaaagga gagggccaag
      421 agggagggca ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag
      481 cttggagtat gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg
      541 gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta
      601 ttatatgtgt gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa
      661 gtcttgtaaa agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc
      721 gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc
      781 aactgcatgg agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta
      841 aaagactcgt atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc
      901 attcgtttat tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga
      961 actcgatttc tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag
     1021 ctggtggact gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg
     1081 ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt
     1141 tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt
     1201 gcacttgccg atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     1261 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     1321 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     1381 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     1441 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     1501 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     1561 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     1621 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     1681 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     1741 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     1801 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     1861 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
     1921 gggccccccc tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg
     1981 ggatccacta gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta
     2041 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2101 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     2161 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2221 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2281 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2341 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2401 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2461 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2521 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2581 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2641 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2701 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2761 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2821 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2881 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2941 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3001 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3061 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3121 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3181 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3241 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3301 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3361 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3421 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3481 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3541 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3601 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3661 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3721 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3781 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3841 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3901 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3961 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4021 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4081 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4141 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4201 cacatttccc cgaaaagtgc cacctgacgt cctaggcatg caacgaagca tctgtgcttc
     4261 attttgtaga acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct
     4321 gcatttttac agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg
     4381 cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     4441 gagctgcatt tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat
     4501 ctatacttct tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc
     4561 atcttagatt actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt
     4621 tgcactgtag gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat
     4681 aaaaaaagcc tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt
     4741 ttttcaagat aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg
     4801 tgaacagaaa gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct
     4861 tctattttgt ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat
     4921 tcactctatg aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa
     4981 cataaaaaat gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta
     5041 ggttatatag ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg
     5101 tggaagcggt attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt
     5161 tgaaagtgcg tcttcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt
     5221 ctagagaata ggaacttcgg aataggaact tcaaagcgtt tccgaaaacg agcgcttccg
     5281 aaaatgcaac gcgagctgcg cacatacagc tcactgttca cgtcgcacct atatctgcgt
     5341 gttgcctgta tatatatata catgagaaga acggcatagt gcgtgtttat gcttaaatgc
     5401 gtacttatat gcgtctattt atgtaggatg aaaggtagtc tagtacctcc tgtgatatta
     5461 tcccattcca tgcggggtat cgtatgcttc cttcagcact accctttagc tgttctatat
     5521 gctgccactc ctcaattgga ttagtctcat ccttcaatgc tatcatttcc tttgatattg
     5581 gatcgcatgc ctaggacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5641 ggcgtatc
//
