LOCUS       sequence-350278-        4840 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast shuttle plasmid that allows conversion between low copy, high 
            copy, and integrating vectors. Low copy, TRP1-selected..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4840)
  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 4840)
  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: pDN514
FEATURES             Location/Qualifiers
     source          1..4840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     84..102
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(140..162)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     262..281
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        302..583
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             584..1258
                     /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(1361..1816)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1498..1519)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1710..1729
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1793..2173)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1942..1964
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1956..1973
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1957..1973
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1983..2002
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1983..2001
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2010..2117
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2026..2042
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2027..2043
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2077..2093)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2077..2093)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2130..2150)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2130..2148)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2169..2185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2169..2185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2182..2204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2193..2209
                     /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(2217..2223,2224..2241,2242..2247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2262..2283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2400..2417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2571..3159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2651..2670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3330..4121,4122..4190))
                     /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     3953..3972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4191..4295)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(4347..4840,1..9)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 caagcgatcc gtctaagaaa ccattattat cgcatgccta ggacgtctaa gaaaccatta
       61 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
      121 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
      181 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
      241 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatagat
      301 caacgacatt actatatata taatatagga agcatttaat agaacagcat cgtaatatat
      361 gtgtactttg cagttatgac gccagatggc agtagtggaa gatattcttt attgaaaaat
      421 agcttgtcac cttacgtaca atcttgatcc ggagcttttc tttttttgcc gattaagaat
      481 taattcggtc gaaaaaagaa aaggagaggg ccaagaggga gggcattggt gactattgag
      541 cacgtgagta tacgtgatta agcacacaaa ggcagcttgg agtatgtctg ttattaattt
      601 cacaggtagt tctggtccat tggtgaaagt ttgcggcttg cagagcacag aggccgcaga
      661 atgtgctcta gattccgatg ctgacttgct gggtattata tgtgtgccca atagaaagag
      721 aacaattgac ccggttattg caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag
      781 ttcaggcact ccgaaatact tggttggcgt gtttcgtaat caacctaagg aggatgtttt
      841 ggctctggtc aatgattacg gcattgatat cgtccaactg catggagatg agtcgtggca
      901 agaataccaa gagttcctcg gtttgccagt tattaaaaga ctcgtatttc caaaagactg
      961 caacatacta ctcagtgcag cttcacagaa acctcattcg tttattccct tgtttgattc
     1021 agaagcaggt gggacaggtg aacttttgga ttggaactcg atttctgact gggttggaag
     1081 gcaagagagc cccgaaagct tacattttat gttagctggt ggactgacgc cagaaaatgt
     1141 tggtgatgcg cttagattaa atggcgttat tggtgttgat gtaagcggag gtgtggagac
     1201 aaatggtgta aaagactcta acaaaatagc aaatttcgtc aaaaatgcta agaaataggt
     1261 tattactgag tagtatttat ttaagtattg tttgtgcact tgccgatcta tgcggtgtga
     1321 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt
     1381 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1441 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1501 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1561 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1621 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1681 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1741 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1801 ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1861 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     1921 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     1981 cgtaatacga ctcactatag ggcgaattgg gtaccgggcc ccccctcgag gtcgacggta
     2041 tcgataagct tgatatcgaa ttcctgcagc ccgggggatc cactagttct agagcggccg
     2101 ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     2161 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2221 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2281 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2341 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2401 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2461 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2521 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2581 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2641 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2701 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2761 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2821 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2881 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2941 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3001 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3061 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3121 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3181 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3241 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3301 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3361 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3421 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3481 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3541 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3601 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3661 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3721 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3781 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3841 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3901 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3961 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4021 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4081 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4141 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4201 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4261 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4321 gacgtcctag gcatgcggtc cttttcatca cgtgctataa aaataattat aatttaaatt
     4381 ttttaatata aatatataaa ttaaaaatag aaagtaaaaa aagaaattaa agaaaaaata
     4441 gtttttgttt tccgaagatg taaaagactc tagggggatc gccaacaaat actacctttt
     4501 atcttgctct tcctgctctc aggtattaat gccgaattgt ttcatcttgt ctgtgtagaa
     4561 gaccacacac gaaaatcctg tgattttaca ttttacttat cgttaatcga atgtatatct
     4621 atttaatctg cttttcttgt ctaataaata tatatgtaaa gtacgctttt tgttgaaatt
     4681 ttttaaacct ttgtttattt tttttcttca ttccgtaact cttctacctt ctttatttac
     4741 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
     4801 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg
//
