LOCUS       Exported                5742 bp DNA     circular SYN 16-AUG-2021
DEFINITION  Yeast shuttle vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5742)
  AUTHORS   Daniel P. Nickerson, Monique A. Quinn, Joshua M. Milnes
  TITLE     Rapid conversion of replicating and integrating Saccharomyces 
            cerevisiae plasmid vectors via Cre recombinase
REFERENCE   2  (bases 1 to 5742)
  AUTHORS   Nickerson Lab
  TITLE     Direct Submission
  JOURNAL   Exported Aug 18, 2021 from SnapGene 5.3.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Preprint"; archive: "bioR-chi-iv"; doi: 
            "https://doi.org/10.1101/2020.11.03.367219"; datePosted: "2020"
FEATURES             Location/Qualifiers
     source          1..5742
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        198..413
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             414..1217
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /note="color: #ff7f50"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1348..1803)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     CDS             complement(1586..2160)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /note="color: #993366"
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWTRPVAAH*ARRVWWLRAA*PLHLPAP*RPLLSLSSLPFSPRSPA
                     FPVKL*IGGSL*GSDLVLYGTSTPKNLI"
     primer_bind     1944..1960
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        1970..1988
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    1997..2104
                     /label=MCS
                     /note="pBluescript multiple cloning site"
                     /note="color: #99ccff"
     promoter        complement(2117..2135)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(2156..2172)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    2180..2196
                     /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)."
                     /note="color: #31849b"
     promoter        complement(2204..2234)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 2204 .. 2210 / #ffffff / -10
                        2: 2211 .. 2228 / #ffffff
                        3: 2229 .. 2234 / #ffffff / -35"
     rep_origin      complement(2558..3146)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(3317..4177)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3317 .. 4108 / #ccffcc
                        2: 4109 .. 4177 / #ccffcc / signal sequence
                     Cleavage site after base 4108"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4178..4282)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    4308..4323
                     /label=AatII-AvrII-SphI
                     /note="color: #ffffff"
     rep_origin      4309..5666
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
                     /note="color: #ffff00"
     misc_feature    4593..4654
                     /label=STB-P
                     /note="Proximal domain of 2µ STB partitioning locus"
                     /note="color: #a6acb3"
     protein_bind    complement(5287..5334)
                     /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)."
                     /note="color: #31849b; direction: LEFT"
     misc_feature    5667..5682
                     /label=AatII-AvrII-SphI
                     /note="color: #ffffff"
ORIGIN
        1 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
       61 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
      121 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
      181 gtgcaccata ccaccttttc aattcatcat ttttttttta ttcttttttt tgatttcggt
      241 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
      301 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
      361 tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa atcatgtcga
      421 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
      481 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
      541 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
      601 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
      661 agtacaattt tttactcttc gaagacagaa aatttgctga cattggtaat acagtcaaat
      721 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
      781 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
      841 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
      901 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
      961 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     1021 gtgtgggttt agatgacaag ggagacgcat tgggtcaaca gtatagaacc gtggatgatg
     1081 tggtctctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     1141 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     1201 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     1261 ttagagcttc aatttaatta tatcagttat taccctatgc ggtgtgaaat accgcacaga
     1321 tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg
     1381 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     1441 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     1501 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     1561 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     1621 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     1681 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     1741 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     1801 cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     1861 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     1921 agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt aatacgactc
     1981 actatagggc gaattgggta ccgggccccc cctcgaggtc gacggtatcg ataagcttga
     2041 tatcgaattc ctgcagcccg ggggatccac tagttctaga gcggccgcca ccgcggtgga
     2101 gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     2161 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2221 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2281 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2341 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2401 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2461 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2521 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2581 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2641 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2701 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2761 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2821 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2881 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2941 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3001 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3061 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3121 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3181 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3241 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3301 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3361 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3421 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3481 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3541 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3601 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3661 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3721 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3781 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3841 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3901 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3961 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4021 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4081 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4141 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4201 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4261 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcctaggca
     4321 tgcaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     4381 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     4441 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     4501 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     4561 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     4621 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     4681 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     4741 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     4801 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     4861 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     4921 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     4981 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     5041 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     5101 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     5161 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     5221 gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag
     5281 cgctctgaag ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaaagcg
     5341 tttccgaaaa cgagcgcttc cgaaaatgca acgcgagctg cgcacataca gctcactgtt
     5401 cacgtcgcac ctatatctgc gtgttgcctg tatatatata tacatgagaa gaacggcata
     5461 gtgcgtgttt atgcttaaat gcgtacttat atgcgtctat ttatgtagga tgaaaggtag
     5521 tctagtacct cctgtgatat tatcccattc catgcggggt atcgtatgct tccttcagca
     5581 ctacccttta gctgttctat atgctgccac tcctcaattg gattagtctc atccttcaat
     5641 gctatcattt cctttgatat tggatcgcat gcctaggacg tctaagaaac cattattatc
     5701 atgacattaa cctataaaaa taggcgtatc acgaggccct tt
//
