LOCUS       40924_30790        5363 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMGK, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5363)
  AUTHORS   Boel G, Letso R, Neely H, Price WN, Wong KH, Su M, Luff JD, Valecha 
            M, Everett JK, Acton TB, Xiao R, Montelione GT, Aalberts DP, Hunt 
            JF.
  TITLE     Codon influence on protein expression in E. coli correlates with 
            mRNA levels
  JOURNAL   Nature 529, 358-363 (2016)
  PUBMED    26760206
REFERENCE   2  (bases 1 to 5363)
  AUTHORS   Boel G, Hunt JF, Montelione GT, Acton T.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JUN-2015) Biological Sciences, Columbia University, 
            1212 Amsterdam Ave., New York, NY 10027, USA
REFERENCE   3  (bases 1 to 5363)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5363)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature 529,
            358-363 (2016)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-JUN-2015) Biological Sciences, Columbia University, 1212 
            Amsterdam Ave., New York, NY 10027, 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..5363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     tRNA            complement(893..969)
                     /label=argU
     promoter        1694..1771
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1772..2851
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    2867..2888
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     terminator      3056..3090
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(3802..4614)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(join(5209..5363,1..391))
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
ORIGIN
        1 ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca
       61 gttcgctcca agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta
      121 tccggtaact atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca
      181 gccactggta attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac
      241 tgaaaggaca agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt
      301 ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca
      361 agagattacg cgcagaccaa aacgatctca agaagatcat cttattaagg ggtctgacgc
      421 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
      481 cacctagatc ctttttcaaa taggcaagca tcacaaatgg gtgtttgtac ataccaaggc
      541 ggctaagcga gcagatggaa catcaacgcc tgcggtcagg aagatgcgca tcgacagcaa
      601 gacatcatgg ctatcagctt gtcgtcgtgc aggaattgaa gatttccgtt tccatgacct
      661 cagacacacc tgggcaagct ggctgattca gtcaggcgtc ccattatcag tgcttcagga
      721 aatgggcgga tgggagtcca tagaaatggt tcgtaggtat gctcaccttg cgcctaatca
      781 tttgacagag catgcgagga aaatagacga catttttggt gataatgtcc caaatatgtc
      841 ccactctgaa attatggagg atataaagaa ggcgtaactg attgaattgt aatggcgcgc
      901 cctgcaggat tcgaacctgc ggcccacgac ttagaaggtc gttgctctat ccaactgagc
      961 taagggcgcg ttgataccgc aatgcggtgt aatcgcgtga attatacggt caacccttgc
     1021 tgagtcaatg gcttttgaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     1081 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1141 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1201 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1261 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1321 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1381 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1441 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1501 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1561 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1621 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1681 cttgcccggc gtcgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg
     1741 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     1801 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     1861 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     1921 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     1981 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     2041 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     2101 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     2161 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     2221 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     2281 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     2341 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     2401 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     2461 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     2521 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     2581 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     2641 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     2701 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     2761 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     2821 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     2881 tcactcatta ggcaccccag gctttacact ttatgcttcc gggttttctt tgcctcacga
     2941 tcgcccccaa aacacataac caattgtatt tattgaaaaa taaatagata caactcacta
     3001 aacatagcaa ttcagatccg gtccagtaat gacctcagaa ctccatctgg atttgttcag
     3061 aacgctcggt tgccgccggg cgttttttat tggtgagaat cgcagcaact tgtcgcgcca
     3121 atcgagccat gtcgtcgtca acgacccccc attcaagaac agcaagcagc attgagaact
     3181 ttggaatcca gtccctcttc cacctgctga ccggatcctc cggcgttcag cctgtgccac
     3241 agccgacagg atggtgacca ccatttgccc catatcaccg tcggtactga tcccgtcgtc
     3301 aataaaccga accgctacac cctgagcatc aaactctttt atcagttgga tcatgtcggc
     3361 ggtgtcgcgg ccaagacggt cgagcttctt caccagaatg acatcacctt cctccacctt
     3421 catcctcagc aaatccagcc cttcccgatc tgttgaactg ccggatgcct tgtcggtaaa
     3481 gatgcggtta gcttttaccc ctgcatcttt gagcgctgag gtctgcctcg tgaagaaggt
     3541 gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggagcca
     3601 cggttgatga gagctttgtt gtaggtggac cagttggtga ttttgaactt ttgctttgcc
     3661 acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt
     3721 cgatttattc aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca
     3781 acaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     3841 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     3901 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     3961 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     4021 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
     4081 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     4141 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     4201 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     4261 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     4321 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     4381 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     4441 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     4501 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     4561 gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcataacacc
     4621 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
     4681 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac
     4741 ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc gttccgtggc
     4801 aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca tcaaccgtgg
     4861 ctccctcact ttctggctgg atgatggggc gattcaggcc tggtatgagt cagcaacacc
     4921 ttcttcacga ggcagacctc agcgctagcg gagtgtatac tggcttacta tgttggcact
     4981 gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
     5041 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
     5101 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     5161 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     5221 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     5281 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     5341 tgcctttcgg tttaccggtg tca
//
