LOCUS       40924_37783        5779 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRS304 vYFPdel-5-1.2(MS2), complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5779)
  AUTHORS   Belmont BJ, Niles JC.
  TITLE     Inducible Control of Subcellular RNA Localization Using a Synthetic 
            Protein-RNA Aptamer Interaction
  JOURNAL   PLoS ONE 7 (10), E46868 (2012)
  PUBMED    23056498
REFERENCE   2  (bases 1 to 5779)
  AUTHORS   Belmont BJ, Niles JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-SEP-2012) Biological Engineering, Massachusetts 
            Institute of Technology, 77 Massachusetts Ave, Cambridge, MA 02139, 
            USA
REFERENCE   3  (bases 1 to 5779)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5779)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2012"; volume: "7"; issue: "10"; pages: "E46868"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-SEP-2012) Biological Engineering, Massachusetts Institute of 
            Technology, 77 Massachusetts Ave, Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        185..465
                     /label=TRP1 promoter
     CDS             466..1137
                     /codon_start=1
                     /label=TRP1
                     /note="phosphoribosylanthranilate isomerase, required for 
                     tryptophan biosynthesis"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1237..1692)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     1837..1853
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1860..1878
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2277..2948)
                     /codon_start=1
                     /label=(3-F)Tyr-EGFP
                     /note="(3-F)Tyr-EGFP is a basic (constitutively
                     fluorescent) green fluorescent protein published in 2004, 
                     derived from Aequorea victoria."
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     ICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLEF
                     VT"
     promoter        complement(3519..3537)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3558..3574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3582..3598)
                     /label=lac operator
                     /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(3606..3636)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3651..3672)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3960..4548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4722..5579)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5580..5684)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accaaacgac attactatat atataatata ggaagcattt aatagacagc atcgtaatat
      241 atgtgtactt tgcagttatg acgccagatg gcagtagtgg aagatattct ttattgaaaa
      301 atagcttgtc accttacgta caatcttgat ccggagcttt tctttttttg ccgattaaga
      361 attaattcgg tcgaaaaaag aaaaggagag ggccaagagg gagggcattg gtgactattg
      421 agcacgtgag tatacgtgat taagcacaca aaggcagctt ggagtatgtc tgttattaat
      481 ttcacaggta gttctggtcc attggtgaaa gtttgcggct tgcagagcac agaggccgca
      541 gaatgtgctc tagattccga tgctgacttg ctgggtatta tatgtgtgcc caatagaaag
      601 agaacaattg acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat
      661 agttcaggca ctccgaaata cttggttggc gtgtttcgta atcaacctaa ggaggatgtt
      721 ttggctctgg tcaatgatta cggcattgat atcgtccaac tgcatggaga tgagtcgtgg
      781 caagaatacc aagagttcct cggtttgcca gttattaaaa gactcgtatt tccaaaagac
      841 tgcaacatac tactcagtgc agcttcacag aaacctcatt cgtttattcc cttgtttgat
      901 tcagaagcag gtgggacagg tgaacttttg gattggaact cgatttctga ctgggttgga
      961 aggcaagaga gccccgaaag cttacatttt atgttagctg gtggactgac gccagaaaat
     1021 gttggtgatg cgcttagatt aaatggcgtt attggtgttg atgtaagcgg aggtgtggag
     1081 acaaatggtg taaaagactc taacaaaata gcaaatttcg tcaaaaatgc taagaaatag
     1141 gttattactg agtagtattt atttaagtat tgtttgtgca cttgcctgcg gtgtgaaata
     1201 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaaacgtt aatattttgt
     1261 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     1321 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     1381 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     1441 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     1501 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     1561 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     1621 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     1681 tacagggcgc gtcgcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1741 tgcgggcctc ttcgctatta cgccagctgg cgaagggggg atgtgctgca aggcgattaa
     1801 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt
     1861 aatacgactc actatagggc gaattggagc tcaagcttta acgaacgcag aattttcgag
     1921 ttattaaact taaaatacgc tgaacccgaa catagaaata tcgaatggga aaaaaaaact
     1981 gcataaaggc attaaaagag gagcgaattt ttttttaata aaaatcttaa taatcattaa
     2041 aagataaata atagtctata tatacgtata taaataaaaa atattcaaaa aataaaataa
     2101 actattattt tagcgtaaag gatggggaaa gagaaaagaa aaaaattgat ctatcgaagg
     2161 aattctaatg aacccgggaa tactgcaggg atccaggcca tcacattccg tccgtatcga
     2221 cctttctctg cctggatcct ttctaggcaa ttaggtacct taggatctct agattaagta
     2281 acaaattcta acaagaccat gtggtctctc ttttcgtttg gatctttgga taaggcagat
     2341 tgataggata agtaatggtt gtctggtaac aagactggac catcaccaat tggagtattt
     2401 tgttgataat ggtcagctaa ttgaacacca ccatcttcaa tgttgtgtct aattttgaag
     2461 ttagctttga taccattctt ttgtttgtca gcagtgatgt aaacattgtg agagttatag
     2521 ttgtattcca atttgtgacc taaaatgtta ccatcttctt taaaatcaat accttttaat
     2581 tcgattctat taactaaggt atcaccttca aacttgactt cagctctggt cttgtagtta
     2641 ccgtcatctt tgaaaaaaat agttctttct tgaacataac cttctggcat ggcagacttg
     2701 aaaaagtcat gttgtttcat atgatctggg tatctagcaa aacacattaa accataacct
     2761 aaagtagtga ctaaggttgg ccatggaact ggcaatttac cagtagtaca aattaatttt
     2821 aaggtcaatt taccgtaagt agcatcacct tcaccttcac cggagacaga aaatttgtga
     2881 ccattaacat caccatctaa ttcaaccaaa attgggacaa caccagtgaa taattcttca
     2941 cctttagaca tatgttctcg agttttgtgt tcttaagtag ataattggat cctttctaat
     3001 gaacaagaag taaaaaaagt tgtaatagaa caagaaaaat gaaactgaaa cttgagaaat
     3061 tgaagaccgt ttattaactt aaatatcaat gggaggtcat cgaaagagaa aaaaatcaaa
     3121 aaaaaaaatt ttcaagaaaa agaaacgtga taaaaatttt tattgccttt ttcgacgaag
     3181 aaaaagaaac gaggcggtct cttttttctt ttccaaacct ttagtacggg taattaacga
     3241 caccctagag gaagaaagag gggaaattta gtatgctgtg cttgggtgtt ttgaagtggt
     3301 acggcgatgc gcggagtccg agaaaatctg gaagagtaaa aaaggagtag aaacattttg
     3361 aagctatggt gtgtggggga tcacttgtgg gggattgggt gtgatgtaag gattcgcggt
     3421 cctcgaaaat taaaagtcca acgcgcctgt tgcttcctat gtgatatgta ttatatgtaa
     3481 tatgcgtcga gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaattccg
     3541 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3601 ccacacaaca taggagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagg
     3661 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3721 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3781 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3841 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3901 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3961 ttccataggc tcggcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4021 cgaaacccga caggactata aagataccag gcgttccccc ctggaagctc cctcgtgcgc
     4081 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4141 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4201 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4261 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4321 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4381 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4441 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4501 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4561 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4621 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4681 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4741 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact gcccgtcgtg
     4801 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4861 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4921 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4981 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5041 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5101 aggcgagtta catgatcccc catgttgtga aaaaaagcgg ttagctcctt cggtcctccg
     5161 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5221 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5281 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     5341 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     5401 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     5461 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5521 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5581 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5641 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5701 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     5761 atcacgaggc cctttcgtc
//
