LOCUS       sequence-350315-        5874 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 5874)
  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 5874)
  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: pDN623
FEATURES             Location/Qualifiers
     source          1..5874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    40..73
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     110..128
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(166..188)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     288..307
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        454..641
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             642..1304
                     /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(1564..2019)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1701..1722)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1913..1932
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1996..2376)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2145..2167
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2159..2176
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2160..2176
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2186..2205
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2186..2204
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2213..2320
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2229..2245
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2230..2246
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2280..2296)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2280..2296)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2333..2353)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2333..2351)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2372..2388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2372..2388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2385..2407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2396..2412
                     /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(2420..2426,2427..2444,2445..2450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2465..2486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2603..2620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2774..3362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2854..2873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3533..4324,4325..4393))
                     /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     4156..4175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4394..4498)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    4524..4557
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      join(4565..5874,1..33)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5528..5575)
                     /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 cttcaatgct atcatttcct ttgatattgg atcgacgtca taacttcgta taatgtatgc
       61 tatacgaagt tattaagaaa ccattattat catgacatta acctataaaa ataggcgtat
      121 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
      181 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
      241 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
      301 gattgtactg agagtgcacc ataaattccc gttttaagag cttggtgagc gctaggagtc
      361 actgccaggt atcgtttgaa cacggcatta gtcagggaag tcataacaca gtcctttccc
      421 gcaattttct ttttctatta ctcttggcct cctctagtac actctatatt tttttatgcc
      481 tcggtaatga ttttcatttt tttttttcca cctagcggat gactcttttt ttttcttagc
      541 gattggcatt atcacataat gaattataca ttatataaag taatgtgatt tcttcgaaga
      601 atatactaaa aaatgagcag gcaagataaa cgaaggcaaa gatgacagag cagaaagccc
      661 tagtaaagcg tattacaaat gaaaccaaga ttcagattgc gatctcttta aagggtggtc
      721 ccctagcgat agagcactcg atcttcccag aaaaagaggc agaagcagta gcagaacagg
      781 ccacacaatc gcaagtgatt aacgtccaca caggtatagg gtttctggac catatgatac
      841 atgctctggc caagcattcc ggctggtcgc taatcgttga gtgcattggt gacttacaca
      901 tagacgacca tcacaccact gaagactgcg ggattgctct cggtcaagct tttaaagagg
      961 ccctaggggc cgtgcgtgga gtaaaaaggt ttggatcagg atttgcgcct ttggatgagg
     1021 cactttccag agcggtggta gatctttcga acaggccgta cgcagttgtc gaacttggtt
     1081 tgcaaaggga gaaagtagga gatctctctt gcgagatgat cccgcatttt cttgaaagct
     1141 ttgcagaggc tagcagaatt accctccacg ttgattgtct gcgaggcaag aatgatcatc
     1201 accgtagtga gagtgcgttc aaggctcttg cggttgccat aagagaagcc acctcgccca
     1261 atggtaccaa cgatgttccc tccaccaaag gtgttcttat gtagtgacac cgattattta
     1321 aagctgcagc atacgatata tatacatgtg tatatatgta tacctatgaa tgtcagtaag
     1381 tatgtatacg aacagtatga tactgaagat gacaaggtaa tgcatcattc tatacgtgtc
     1441 attctgaacg aggcgcgctt tccttttttc tttttgcttt ttcttttttt ttctcttgaa
     1501 ctcgacggat ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     1561 aggaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     1621 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     1681 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     1741 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     1801 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     1861 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     1921 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     1981 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg
     2041 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     2101 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     2161 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
     2221 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg
     2281 atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
     2341 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2401 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2461 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2521 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2581 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2941 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4501 catttccccg aaaagtgcca cctataactt cgtataatgt atgctatacg aagttattga
     4561 cgtcaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
     4621 ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
     4681 attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
     4741 ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
     4801 gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
     4861 gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
     4921 aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
     4981 ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
     5041 actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
     5101 cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
     5161 tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
     5221 tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
     5281 taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
     5341 aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
     5401 gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
     5461 agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa
     5521 gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga acttcaaagc
     5581 gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt
     5641 tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat
     5701 agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg atgaaaggta
     5761 gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc
     5821 actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct catc
//
