LOCUS       sequence-350317-        5697 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 5697)
  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 5697)
  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: pDN624
FEATURES             Location/Qualifiers
     source          1..5697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    29..62
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     99..117
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(155..177)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     277..296
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        317..598
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             599..1273
                     /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(1376..1831)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1513..1534)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1725..1744
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(1808..2188)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1957..1979
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1971..1988
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1972..1988
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1998..2017
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1998..2016
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2025..2132
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     2041..2057
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2042..2058
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2092..2108)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2092..2108)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2145..2165)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2145..2163)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2184..2200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2184..2200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2197..2219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2208..2224
                     /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(2232..2238,2239..2256,2257..2262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2277..2298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2415..2432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2586..3174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2666..2685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3345..4136,4137..4205))
                     /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     3968..3987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4206..4310)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    4336..4369
                     /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(4377..5697,1..22)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(5340..5387)
                     /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 tcatttcctt tgatattgga tcgacgtcat aacttcgtat aatgtatgct atacgaagtt
       61 attaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
      121 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
      181 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
      241 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
      301 gagtgcacca tagatcaacg acattactat atatataata taggaagcat ttaatagaac
      361 agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag tggaagatat
      421 tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc ttttcttttt
      481 ttgccgatta agaattaatt cggtcgaaaa aagaaaagga gagggccaag agggagggca
      541 ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag cttggagtat
      601 gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg gcttgcagag
      661 cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta ttatatgtgt
      721 gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa gtcttgtaaa
      781 agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc gtaatcaacc
      841 taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc aactgcatgg
      901 agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta aaagactcgt
      961 atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc attcgtttat
     1021 tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga actcgatttc
     1081 tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag ctggtggact
     1141 gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg ttgatgtaag
     1201 cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt tcgtcaaaaa
     1261 tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt gcacttgccg
     1321 atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     1381 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1441 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     1501 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     1561 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     1621 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     1681 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     1741 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     1801 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     1861 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     1921 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     1981 cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc gggccccccc
     2041 tcgaggtcga cggtatcgat aagcttgata tcgaattcct gcagcccggg ggatccacta
     2101 gttctagagc ggccgccacc gcggtggagc tccagctttt gttcccttta gtgagggtta
     2161 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2221 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2281 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2341 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2401 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2461 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2521 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2581 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2641 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2701 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2761 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2821 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2881 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2941 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3001 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3061 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3121 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3181 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3241 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3301 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3361 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3421 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3481 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3541 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3601 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3661 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3721 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3781 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3841 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3901 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3961 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4021 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4081 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4141 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4201 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4261 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4321 cgaaaagtgc cacctataac ttcgtataat gtatgctata cgaagttatt gacgtcaacg
     4381 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa
     4441 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc
     4501 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt
     4561 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt
     4621 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta
     4681 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt
     4741 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc
     4801 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga
     4861 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg
     4921 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa
     4981 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt
     5041 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt
     5101 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg
     5161 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact
     5221 tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt
     5281 gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg
     5341 aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga
     5401 aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg
     5461 cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg
     5521 tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta
     5581 cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct
     5641 ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgcta
//
