LOCUS       40924_49467        7428 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector YCplac22 5-1.2-FLuciferase, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7428)
  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 7428)
  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 7428)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7428)
  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..7428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..33
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             678..2327
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2632..2648)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(3407..4569)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     promoter        complement(5083..5184)
                     /label=TRP1 promoter
     promoter        5290..5394
                     /label=AmpR promoter
     CDS             5395..6252
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6426..7014
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7302..7323
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7338..7368
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7376..7392
                     /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)."
     primer_bind     7400..7416
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcccagtcac gacgttgtaa aacgacggcc agtgctacaa tcgcggccgc ataggtatcg
       61 atcgtcaccc tgcagggtga cggatcgatc cgtcacccgc atattacata taatacatat
      121 cacataggaa gcaacaggcg cgttggactt ttaattttcg aggaccgcga atccttacat
      181 cacacccaat cccccacaag tgatccccca cacaccatag cttcaaaatg tttctactcc
      241 ttttttactc ttccagattt tctcggactc cgcgcatcgc cgtaccactt caaaacaccc
      301 aagcacagca tactaaattt cccctctttc ttcctctagg gtgtcgttaa ttacccgtac
      361 taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt tttcttcgtc gaaaaaggca
      421 ataaaaattt ttatcacgtt tctttttctt gaaaattttt ttttttgatt tttttctctt
      481 tcgatgacct cccattgata tttaagttaa taaacggtct tcaatttctc aagtttcagt
      541 ttcatttttc ttgttctatt acaacttttt ttacttcttg ttcattagaa aggatctaat
      601 tatctaggat ccaggcagag aaaggtcgat acggacggaa tgtgatggcc tggatccaac
      661 acaaaactcg agaacatatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct
      721 atccactaga ggatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc
      781 tggttcctgg aacaattgct tttacagatg cacatatcga ggtgaacatc acgtacgcgg
      841 aatacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa
      901 atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg
      961 cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc
     1021 tcaacagtat gaacatttcg cagcctaccg tagtgtttgt ttccaaaaag gggttgcaaa
     1081 aaattttgaa cgtgcaaaaa aaattaccaa taatccagaa aattattatc atggattcta
     1141 aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg
     1201 gttttaatga atacgatttt gtaccagagt cctttgatcg tgacaaaaca attgcactga
     1261 taatgaattc ctctggatct actgggttac ctaagggtgt ggcccttccg catagaactg
     1321 cctgcgtcag attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata
     1381 ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat
     1441 atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttttac
     1501 gatcccttca ggattacaaa attcaaagtg cgttgctagt accaacccta ttttcattct
     1561 tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg
     1621 ggggcgcacc tctttcgaaa gaagtcgggg aagcggttgc aaaacgcttc catcttccag
     1681 ggatacgaca aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg
     1741 gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg
     1801 atctggatac cgggaaaacg ctgggcgtta atcagagagg cgaattatgt gtcagaggac
     1861 ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg
     1921 atggatggct acattctgga gacatagctt actgggacga agacgaacac ttcttcatag
     1981 ttgaccgctt gaagtcttta attaaataca aaggatatca ggtggccccc gctgaattgg
     2041 aatcgatatt gttacaacac cccaacatct tcgacgcggg cgtggcaggt cttcccgacg
     2101 atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg
     2161 aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag
     2221 gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa
     2281 tcagagagat cctcataaag gccaagaagg gcggaaagtc caaattgtaa tctagaattc
     2341 cttcgataga tcaatttttt tcttttctct ttccccatcc tttacgctaa aataatagtt
     2401 tattttattt tttgaatatt ttttatttat atacgtatat atagactatt atttatcttt
     2461 taatgattat taagattttt attaaaaaaa aattcgctcc tcttttaatg cctttatgca
     2521 gttttttttt cccattcgat atttctatgt tcgggttcag cgtattttaa gtttaataac
     2581 tcgaaaattc tgcgttcgtt aaagcttggc gtaatcatgg tcatagctgt tactggccgt
     2641 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     2701 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     2761 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     2821 gtgcggtatt tcacaccgca tatatcgctg ggccattctc atgaagaata tcttgaattt
     2881 attgtcatat tactagttgg tgtggaagtc catatatcgg tgatcaatat agtggttgac
     2941 atgctggcta gtcaacattg agccttttga tcatgcaaat atattacggt attttacaat
     3001 caaatatcaa acttaactat tgactttata acttatttag gtggtaacat tcttataaaa
     3061 aagaaaaaaa ttactgcaaa acagtactag cttttaactt gtatcctagg ttatctatgc
     3121 tgtctcacca tagagaatat tacctatttc agaatgtatg tccatgattc gccgggtaaa
     3181 tacatataat acacaaatct ggcttaataa agtctataat atatctcata aagaagtgct
     3241 aaattggcta gtgctatata tttttaagaa aatttctttt gactaagtcc atatcgactt
     3301 tgtaaaagtt cactttagca tacatatatt acacgagcca gaaattgtaa cttttgccta
     3361 aaatcacaaa ttgcaaaatt taattgcttg caaaaggtca catgcttata atcaactttt
     3421 ttaaaaattt aaaatacttt tttatttttt atttttaaac ataaatgaaa taatttattt
     3481 attgtttatg attaccgaaa cataaaacct gctcaagaaa aagaaactgt tttgtccttg
     3541 gaaaaaaagc actacctagg agcggccaaa atgccgaggc tttcatagct taaactcttt
     3601 acagaaaata ggcattatag atcagttcga gttttcttat tcttccttcc ggttttatcg
     3661 tcacagtttt acagtaaata agtatcacct cttagagttc gatgataagc tgtcaaacat
     3721 gagaattaat tccacatgtt aaaatagtga aggagcatgt tcggcacaca gtggaccgaa
     3781 cgtggggtaa gtgcactagg gtccggttaa acggatctcg cattgatgag gcaacgctaa
     3841 ttatcaacat atagattgtt atctatctgc atgaacacga aatctttact tgacgacttg
     3901 aggctgatgg tgtttatgca aagaaaccac tgtgtttaat atgtgtcact gtttgatatt
     3961 actgtcagcg tagaagataa tagtaaaagc ggttaataag tgtatttgag ataagtgtga
     4021 taaagttttt acagcgaaaa gacgataaat acaagaaaat gattacgagg atacggagag
     4081 aggtatgtac atgtgtattt atatactaag ctgccggcgg ttgtttgcaa gaccgagaaa
     4141 aggctagcaa gaatcgggtc attgtagcgt atgcgcctgt gaacattctc ttcaacaagt
     4201 ttgattccat tgcggtgaaa tggtaaaagt caaccccctg cgatgtatat tttcctgtac
     4261 aatcaatcaa aaagccaaat gatttagcat tatctttaca tcttgttatt ttacagattt
     4321 tatgtttaga tcttttatgc ttgcttttca aaaggcttgc aggcaagtgc acaaacaata
     4381 cttaaataaa tactactcag taataaccta tttcttagca tttttgacga aatttgctat
     4441 tttgttagag tcttttacac catttgtctc cacacctccg cttacatcaa caccaataac
     4501 gccatttaat ctaagcgcat caccaacatt ttctggcgtc agtccaccag ctaacataaa
     4561 atgtaagctc tcggggctct cttgccttcc aacccagtca gaaatcgagt tccaatccaa
     4621 aagttcacct gtcccacctg cttctgaatc aaacaaggga ataaacgaat gaggtttctg
     4681 tgaagctgca ctgagtagta tgttgcagtc ttttggaaat acgagtcttt taataactgg
     4741 caaaccgagg aactcttggt attcttgcca cgactcatct ccatgcagtt ggacgatatc
     4801 aatgccgtaa tcattgacca gagccaaaac atcctcctta ggttgattac gaaacacgcc
     4861 aaccaagtat ttcggagtgc ctgaactatt tttatatgct tttacaagac ttgaaatttt
     4921 ccttgcaata accgggtcaa ttgttctctt tctattgggc acacatataa tacccagcaa
     4981 gtcagcatcg gaatctagtg cacattctgc ggcctctgtg ctctgcaagc cgcaaacttt
     5041 caccaatgga ccagaactac ctgtgaaatt aataacagac atactccaag ctgcctttgt
     5101 gtgcttaatc acgtatactc acgtgctcaa tagtcaccaa tgccctccct cttggccctc
     5161 tccttttctt ttttcgaccg aattaattct tgaagacgaa agggcctcgt gatacgccta
     5221 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5281 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5341 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5401 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5461 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5521 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5581 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5641 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5701 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5761 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5821 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5881 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5941 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6001 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6061 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6121 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6181 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6241 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6301 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6361 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6421 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6481 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6541 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     6601 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6661 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6721 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6781 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6841 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6901 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6961 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7021 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7081 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7141 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7201 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     7261 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7321 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7381 agcggataac aatttcacac aggaaacagc tatgaccatg attacgcc
//
