LOCUS       sequence-350311-        4891 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 4891)
  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 4891)
  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: pDN614
FEATURES             Location/Qualifiers
     source          1..4891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    238..271
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     308..326
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(364..386)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     486..505
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        526..807
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             808..1482
                     /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(1585..2040)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1722..1743)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1934..1953
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(2017..2397)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     2166..2188
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2180..2197
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2181..2197
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2207..2226
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2207..2225
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2234..2341
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2250..2266
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2251..2267
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2301..2317)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2301..2317)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2354..2374)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2354..2372)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2393..2409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2393..2409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2406..2428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2417..2433
                     /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(2441..2447,2448..2465,2466..2471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2486..2507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2624..2641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2795..3383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2875..2894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3554..4345,4346..4414))
                     /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     4177..4196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4415..4519)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    4545..4578
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(4614..4891,1..226)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 aatctgcttt tcttgtctaa taaatatata tgtaaagtac gctttttgtt gaaatttttt
       61 aaacctttgt ttattttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt
      121 ctaaaatcca aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc
      181 catcattaaa agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc cgacgtcata
      241 acttcgtata atgtatgcta tacgaagtta ttaagaaacc attattatca tgacattaac
      301 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
      361 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
      421 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
      481 ctatgcggca tcagagcaga ttgtactgag agtgcaccat agatcaacga cattactata
      541 tatataatat aggaagcatt taatagaaca gcatcgtaat atatgtgtac tttgcagtta
      601 tgacgccaga tggcagtagt ggaagatatt ctttattgaa aaatagcttg tcaccttacg
      661 tacaatcttg atccggagct tttctttttt tgccgattaa gaattaattc ggtcgaaaaa
      721 agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg
      781 attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt
      841 ccattggtga aagtttgcgg cttgcagagc acagaggccg cagaatgtgc tctagattcc
      901 gatgctgact tgctgggtat tatatgtgtg cccaatagaa agagaacaat tgacccggtt
      961 attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg cactccgaaa
     1021 tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct ggtcaatgat
     1081 tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc
     1141 ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt
     1201 gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca
     1261 ggtgaacttt tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgaa
     1321 agcttacatt ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga
     1381 ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac
     1441 tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat
     1501 ttatttaagt attgtttgtg cacttgccga tctatgcggt gtgaaatacc gcacagatgc
     1561 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt
     1621 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     1681 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     1741 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     1801 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     1861 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     1921 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     1981 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     2041 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     2101 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     2161 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     2221 atagggcgaa ttgggtaccg ggccccccct cgaggtcgac ggtatcgata agcttgatat
     2281 cgaattcctg cagcccgggg gatccactag ttctagagcg gccgccaccg cggtggagct
     2341 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     2401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     2581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     2641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2701 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2761 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3301 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3361 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3421 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3481 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3541 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3841 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4141 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4201 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     4441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctataact tcgtataatg
     4561 tatgctatac gaagttattg acgtccccga aaagtgccac ctgggtcctt ttcatcacgt
     4621 gctataaaaa taattataat ttaaattttt taatataaat atataaatta aaaatagaaa
     4681 gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag
     4741 ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc
     4801 gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga ttttacattt
     4861 tacttatcgt taatcgaatg tatatctatt t
//
