LOCUS       40924_37698        8799 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRR26, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8799)
  AUTHORS   Ramsden R, Arms L, Davis TN, Muller EG.
  TITLE     An intein with genetically selectable markers provides a new 
            approach to internally label proteins with GFP
  JOURNAL   BMC Biotechnol. 11 (1), 71 (2011)
  PUBMED    21708017
REFERENCE   2  (bases 1 to 8799)
  AUTHORS   Ramsden R, Arms L, Davis TN, Muller EGD.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JUN-2011) Biochemistry, University of Washington, 1705
            NE Pacific St, HSB J461, Seattle, WA 98195, USA
REFERENCE   3  (bases 1 to 8799)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8799)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Biotechnol."; date: "2011"; volume: "11"; issue: "1"; pages: "71"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-JUN-2011) Biochemistry, University of Washington, 1705 NE 
            Pacific St, HSB J461, Seattle, WA 98195, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        14..678
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     CDS             694..714
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             724..744
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             748..768
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             790..1500
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     promoter        2289..2509
                     /label=URA3 promoter
     CDS             2510..3310
                     /codon_start=1
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     misc_feature    complement(4658..6507)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     misc_feature    6649..6789
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6975..7563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7737..8594)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8595..8699)
                     /label=AmpR promoter
ORIGIN
        1 gaattccttg aattttcaaa aattcttact ttttttttgg atggacgcaa agaagtttaa
       61 taatcatatt acatggcatt accaccatat acatatccat atacatatcc atatctaatc
      121 ttacttatat gttgtggaaa tgtaaagagc cccattatct tagcctaaaa aaaccttctc
      181 tttggaactt tcagtaatac gcttaactgc tcattgctat attgaagtac ggattagaag
      241 ccgccgagcg ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc tcgtcttcac
      301 cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt
      361 ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa
      421 accttcaaat gaacgaatca aattaacaac cataggatga taatgcgatt agttttttag
      481 ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat
      541 aaatgcaaaa actgcataac cactttaact aatactttca acattttcgg tttgtattac
      601 ttcttattca aatgtaataa aagtatcaac aaaaaattgt taatatacct ctatacttta
      661 acgtcaagga gaaaaaaccc cggatccatg gttccaaaga agaagagaaa ggttttaatt
      721 gatccaaaaa agaaaagaaa ggttgatcca aagaaaaaga gaaaagtttt aattaaagga
      781 gcaggagcaa gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
      841 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca
      901 tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
      961 acacttgtca ctactttcac ttatggtgtt caatgctttt caagataccc agatcatatg
     1021 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
     1081 tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc
     1141 cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga
     1201 cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag
     1261 aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag cgttcaacta
     1321 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
     1381 cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg
     1441 gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa
     1501 tagtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     1561 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     1621 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     1681 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     1741 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
     1801 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
     1861 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
     1921 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
     1981 gatcactgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     2041 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     2101 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
     2161 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
     2221 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatcggg
     2281 gggggggttt caattcaatt catcattttt tttttattct tttttttgat ttcggtttct
     2341 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     2401 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     2461 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
     2521 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
     2581 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
     2641 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
     2701 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
     2761 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
     2821 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
     2881 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
     2941 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
     3001 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
     3061 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     3121 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     3181 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     3241 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     3301 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     3361 agcttcaatt taattatatc agttattacc cgggaatctc ggtcgtaatg atttttataa
     3421 tgacgaaaaa aaaaaaattg gaaagaaaac cccccccccg cagcgttggg tcctggccac
     3481 gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
     3541 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
     3601 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
     3661 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
     3721 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
     3781 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
     3841 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
     3901 tcattacccc catgaacaga aattccccct tacacggagg catcaagtga ccaaacagga
     3961 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
     4021 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     4081 tgatgagctt taccgcaggt gggccattct catgaagaat atcttgaatt tattgtcata
     4141 ttactagttg gtgtggaagt ccatatatcg gtgatcaata tagtggttga catgctggct
     4201 agtcaacatt gagccttttg atcatgcaaa tatattacgg tattttacaa tcaaatatca
     4261 aacttaacta ttgactttat aacttattta ggtggtaaca ttcttataaa aaagaaaaaa
     4321 attactgcaa aacagtacta gcttttaact tgtatcctag gttatctatg ctgtctcacc
     4381 atagagaata ttacctattt cagaatgtat gtccatgatt cgccgggtaa atacatataa
     4441 tacacaaatc tggcttaata aagtctataa tatatctcat aaagaagtgc taaattggct
     4501 agtgctatat atttttaaga aaatttcttt tgactaagtc catatcgact ttgtaaaagt
     4561 tcactttagc atacatatat tacacgagcc agaaattgta acttttgcct aaaatcacaa
     4621 attgcaaaat ttaattgctt gcaaaaggtc acatgcttat aatcaacttt tttaaaaatt
     4681 taaaatactt ttttattttt tatttttaaa cataaatgaa ataatttatt tattgtttat
     4741 gattaccgaa acataaaacc tgctcaagaa aaagaaactg ttttgtcctt ggaaaaaaag
     4801 cactacctag gagcggccaa aatgccgagg ctttcatagc ttaaactctt tacagaaaat
     4861 aggcattata gatcagttcg agttttctta ttcttccttc cggttttatc gtcacagttt
     4921 tacagtaaat aagtatcacc tcttagagtt aactatgaga taagcaagta tcatctcatt
     4981 tcatttacct gaagtcgagt aaacagaaaa tccaattgtt gatgaacctc aatgacttag
     5041 aactatctat cggcagatca tataaagagg atttaggtac ctagaggact gtacctggag
     5101 tatatatata tatatatata tattatctca actatagtcc atagaggttt ctttcttgag
     5161 gccttaaact gctaaagaat gatattggtg gaatgcaagc accaatctct cttctttcgt
     5221 aactgttcat atacttcaaa ccaagaatgt aacgggcatt gacccatcca aaaccttcag
     5281 tagctgcccc tttaaagtca gcaccttgat taccgtattc tgcttcaaca cgatgaggat
     5341 ctgttcctct tgtgacatca tatttttcaa ccacaatacc attataatcg acaaaagcct
     5401 ttgtcatcat gaaaagccat ctataagcta gcctattcgt tacagttaaa taaccataag
     5461 aacggaggcc ttcccaagca agaatttgat ggggtgccca accaaatgga tagtcccatt
     5521 gtctaattgg tctcgaaata gaaattgggc ctcgagaacg ctccgtacat gcagctaaac
     5581 ctccaagcat ctctaacttg ggtagtgctt tctccaccat tttctgtgct tgctccttcg
     5641 tggcaagtcc agcccataat gcccagaatg tagttgcgga ttcgtatgac gttctgtgct
     5701 tgatttttgt gttgtagtca aagaaaaacc ccgactcgtc atcccacata tatttggtaa
     5761 ttgatgaggc aacgctaatt atcaacatat agattgttat ctatctgcat gaacacgaaa
     5821 tctttacttg acgacttgag gctgatggtg tttatgcaaa gaaaccactg tgtttaatat
     5881 gtgtcactgt ttgatattac tgtcagcgta gaagataata gtaaaagcgg ttaataagtg
     5941 tatttgagat aagtgtgata aagtttttac agcgaaaaga cgataaatac aagaaaatga
     6001 ttacgaggat acggagagag gtatgtacat gtgtatttat atactaagct gccggcggtt
     6061 gtttgcaaga ccgagaaaag gctagcaaga atcgggtcat tgtagcgtat gcgcctgtga
     6121 acattctctt caacaagttt gattccattg cggtgaaatg gtaaaagtca accccctgcg
     6181 atgtatattt tcctgtacaa tcaatcaaaa agccaaatga tttagcatta tctttacatc
     6241 ttgttatttt acagatttta tgtttagatc ttttatgctt gcttttcaaa aggcctgcag
     6301 gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt
     6361 tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct
     6421 tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag
     6481 tccaccagct aacataaaat gtaagctctg cctcgcgcgt ttcggtgatg acggtgaaaa
     6541 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     6601 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     6661 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     6721 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     6781 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6841 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6901 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6961 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7021 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7081 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7141 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7201 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7261 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7321 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7381 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7441 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7501 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7561 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7621 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7681 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7741 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7801 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7861 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7921 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7981 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8041 gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8101 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8161 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8221 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8281 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8341 ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8401 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8461 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8521 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8581 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8641 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8701 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     8761 tataaaaata ggcgtatcac gaggcccttt cgtcttcaa
//
