LOCUS       sequence-350276-        5018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast shuttle plasmid that allows conversion between low copy, high 
            copy, and integrating vectors. Low copy, HIS3-selected..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5018)
  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 5018)
  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: pDN513
FEATURES             Location/Qualifiers
     source          1..5018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     269..287
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(325..347)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     447..466
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        613..800
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             801..1463
                     /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(1723..2178)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1860..1881)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2072..2091
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2155..2535)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2304..2326
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2318..2335
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2319..2335
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2345..2364
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2345..2363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2372..2479
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2388..2404
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2389..2405
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2439..2455)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2439..2455)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2492..2512)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2492..2510)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2531..2547)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2531..2547)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2544..2566)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2555..2571
                     /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(2579..2585,2586..2603,2604..2609))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2624..2645
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2762..2779)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2933..3521)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3013..3032)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3692..4483,4484..4552))
                     /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     4315..4334
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4553..4657)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    join(4709..5018,1..194)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg
       61 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat
      121 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt
      181 acaggcaagc gatccgtcta agaaaccatt attatcgcat gcctaggacg tctaagaaac
      241 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
      301 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
      361 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      421 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      481 taaattcccg ttttaagagc ttggtgagcg ctaggagtca ctgccaggta tcgtttgaac
      541 acggcattag tcagggaagt cataacacag tcctttcccg caattttctt tttctattac
      601 tcttggcctc ctctagtaca ctctatattt ttttatgcct cggtaatgat tttcattttt
      661 ttttttccac ctagcggatg actctttttt tttcttagcg attggcatta tcacataatg
      721 aattatacat tatataaagt aatgtgattt cttcgaagaa tatactaaaa aatgagcagg
      781 caagataaac gaaggcaaag atgacagagc agaaagccct agtaaagcgt attacaaatg
      841 aaaccaagat tcagattgcg atctctttaa agggtggtcc cctagcgata gagcactcga
      901 tcttcccaga aaaagaggca gaagcagtag cagaacaggc cacacaatcg caagtgatta
      961 acgtccacac aggtataggg tttctggacc atatgataca tgctctggcc aagcattccg
     1021 gctggtcgct aatcgttgag tgcattggtg acttacacat agacgaccat cacaccactg
     1081 aagactgcgg gattgctctc ggtcaagctt ttaaagaggc cctaggggcc gtgcgtggag
     1141 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
     1201 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
     1261 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
     1321 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
     1381 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
     1441 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
     1501 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
     1561 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
     1621 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt
     1681 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata
     1741 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1801 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1861 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1921 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1981 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2041 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2101 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2161 cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     2221 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     2281 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg
     2341 cgcgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcgacgg
     2401 tatcgataag cttgatatcg aattcctgca gcccggggga tccactagtt ctagagcggc
     2461 cgccaccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     2521 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2581 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2641 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2701 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2761 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2821 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2881 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2941 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3001 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3061 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3121 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3181 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3241 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3301 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3361 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3421 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3481 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3541 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3601 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3661 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3721 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3781 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3841 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3901 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3961 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4021 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4081 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4141 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4201 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4261 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4321 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4381 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4441 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4501 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4561 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4621 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4681 ctgacgtcct aggcatgcgg tccttttcat cacgtgctat aaaaataatt ataatttaaa
     4741 ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa
     4801 tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt
     4861 ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag
     4921 aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat
     4981 ctatttaatc tgcttttctt gtctaataaa tatatatg
//
