LOCUS       40924_28187        6410 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLGFPl DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6410)
  AUTHORS   Kobayashi H.
  TITLE     Inducible suppression of global translation by overuse of rare 
            codons
  JOURNAL   Appl. Environ. Microbiol. 81 (7), 2544-2553 (2015)
  PUBMED    25636849
REFERENCE   2  (bases 1 to 6410)
  AUTHORS   Kobayashi H.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-JAN-2015) Contact:Hideki Kobayashi Agency for 
            Marine-Earth Science and Technology, Institute of Biogeosciences; 
            2-15 Natsushima, Yokosuka, Kanagawa 237-0061, Japan URL 
            :http://www.jamstec.go.jp/j/
REFERENCE   3  (bases 1 to 6410)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6410)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2015"; volume: "81"; issue: "7"; pages:
            "2544-2553"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-JAN-2015) Contact:Hideki Kobayashi Agency for Marine-Earth 
            Science and Technology, Institute of Biogeosciences; 2-15 
            Natsushima, Yokosuka, Kanagawa 237-0061, Japan URL 
            :http://www.jamstec.go.jp/j/"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(22..1101)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             complement(1317..1382)
                     /codon_start=1
                     /label=lambda N peptide
                     /note="N-terminal RNA-binding domain from the lambda
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     promoter        1723..1752
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    1760..1776
                     /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)."
     CDS             1796..2509
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     terminator      2723..2809
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2901..2928
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2948..3039
                     /label=AmpR promoter
     CDS             3040..3897
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3930..4024
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4516..4738
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4786..5733
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(6271..6357)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 aagcttataa ggcgcgcctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
       61 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
      121 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
      181 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
      241 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
      301 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
      361 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
      421 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
      481 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
      541 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
      601 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
      661 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
      721 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
      781 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
      841 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
      901 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
      961 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     1021 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     1081 gtataacgtt actggtttca tgacgtccat ccggccgtcc tttgcataca ccataggtgt
     1141 ggtttaattt gatgcccttt ttcagggctg gaatgtgtaa gagcggggtt atttatgctg
     1201 ttgttttttt gttactcggg aagggcttta cctcttccgc ataaacgctt ccatcagcgt
     1261 ttatagttaa aaaaatcttt cggaactggt tttgcgctta ccccaaccaa caggggattt
     1321 gctgctttcc attgagcctg tttctctgcg cgacgttcgc ggcggcgtgt ttgtgcatcc
     1381 atctggattc tcctgtcagt tagctttggt ggtgtgtggc agttgtagtc ctgaacgaaa
     1441 accccccgcg attggcacat tggcagctaa tccggaatcg cacttacggc caatgcttcg
     1501 tttcgtatca cacaccccaa agccttctgc tttgaatgct gcccttcttc agggcttaat
     1561 ttttaagagc gtcaccttca tggtggtcag tgcgtcctgc tgatgtgctc attataaccg
     1621 ccagtggtat ttatgtcaac accgccagag ataatttatc accgcagatg gttatctgtg
     1681 catgcattta cgttgacacc atcgaatggc tgaaatgagc tgttgacaat taatcatccg
     1741 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa ccggtatgtc
     1801 aaagggggag gagctattca caggggtagt acccatacta gtagagctag acggggacgt
     1861 aaatgggcaa aagttctcag tatcagggga gggggagggg gacgctacat acgggaagct
     1921 aacactaaag ttcatatgta caacagggaa gctacccgta ccctggccca cactagtaac
     1981 aacattcggg tacggggtac aatgtttcgc taggtacccc gaccacatga agcaacacga
     2041 cttcttcaag tcagctatgc ccgaggggta cgtacaagag aggacaatat tctacaagga
     2101 cgacgggaat tacaagacaa gggctgaggt aaagttcgag ggggacacac tagtaaatag
     2161 gatagagcta aaggggatag acttcaagga ggacgggaat atactagggc acaagatgga
     2221 gtacaattac aattcacaca atgtatacat aatggctgac aagcccaaga atgggataaa
     2281 ggtaaatttc aagataaggc acaatataaa ggacgggtca gtacaactag ctgaccacta
     2341 ccaacaaaat acacccatag gggacgggcc cgtactacta cccgacaatc actacctatc
     2401 aacacaatca gctctatcaa aggaccccaa tgagaagagg gaccacatga tactactaga
     2461 gttcgtaaca gctgctggga taacacacgg gatggacgag ctatacaagt aggctagctt
     2521 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
     2581 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     2641 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
     2701 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     2761 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     2821 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     2881 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     2941 aactcttttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     3001 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     3061 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     3121 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3181 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3241 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     3301 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3361 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3421 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3481 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3541 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctacagcaat ggcaacaacg
     3601 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3661 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3721 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3781 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3841 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3901 ctgtcagacc aagtttacga gctcgcttgg actcctgttg atagatccag taatgacctc
     3961 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
     4021 gaatccaagc actagggaca gtaagacggg taagcctgtt gatgataccg ctgccttact
     4081 gggtgcatta gccagtctga atgacctgtc acgggataat ccgaagtggt cagactggaa
     4141 aatcagaggg caggaactgc tgaacagcaa aaagtcagat agcaccacat agcagacccg
     4201 ccataaaacg ccctgagaag cccgtgacgg gcttttcttg tattatgggt agtttccttg
     4261 catgaatcca taaaaggcgc ctgtagtgcc atttaccccc attcactgcc agagccgtga
     4321 gcgcagcgaa ctgaatgtca cgaaaaagac agcgactcag gtgcctgatg gtcggagaca
     4381 aaaggaatat tcagcgattt gcccgagctt gcgagggtgc tacttaagcc tttagggttt
     4441 taaggtctgt tttgtagagg agcaaacagc gtttgcgaca tccttttgta atactgcgga
     4501 actgactaaa gtagtgagtt atacacaggg ctgggatcta ttctttttat ctttttttat
     4561 tctttcttta ttctataaat tataaccact tgaatataaa caaaaaaaac acacaaaggt
     4621 ctagcggaat ttacagaggg tctagcagaa tttacaagtt ttccagcaaa ggtctagcag
     4681 aatttacaga tacccacaac tcaaaggaaa aggactagta attatcattg actagcccat
     4741 ctcaattggt atagtgatta aaatcaccta gaccaattga gatgtatgtc tgaattagtt
     4801 gttttcaaag caaatgaact agcgattagt cgctatgact taacggagca tgaaaccaag
     4861 ctaattttat gctgtgtggc actactcaac cccacgattg aaaaccctac aaggaaagaa
     4921 cggacggtat cgttcactta taaccaatac gctcagatga tgaacatcag tagggaaaat
     4981 gcttatggtg tattagctaa agcaaccaga gagctgatga cgagaactgt ggaaatcagg
     5041 aatcctttgg ttaaaggctt tgagattttc cagtggacaa actatgccaa gttctcaagc
     5101 gaaaaattag aattagtttt tagtgaagag atattgcctt atcttttcca gttaaaaaaa
     5161 ttcataaaat ataatctgga acatgttaag tcttttgaaa acaaatactc tatgaggatt
     5221 tatgagtggt tattaaaaga actaacacaa aagaaaactc acaaggcaaa tatagagatt
     5281 agccttgatg aatttaagtt catgttaatg cttgaaaata actaccatga gtttaaaagg
     5341 cttaaccaat gggttttgaa accaataagt aaagatttaa acacttacag caatatgaaa
     5401 ttggtggttg ataagcgagg ccgcccgact gatacgttga ttttccaagt tgaactagat
     5461 agacaaatgg atctcgtaac cgaacttgag aacaaccaga taaaaatgaa tggtgacaaa
     5521 ataccaacaa ccattacatc agattcctac ctacgtaacg gactaagaaa aacactacac
     5581 gatgctttaa ctgcaaaaat tcagctcacc agttttgagg caaaattttt gagtgacatg
     5641 caaagtaagc atgatctcaa tggttcgttc tcatggctca cgcaaaaaca acgaaccaca
     5701 ctagagaaca tactggctaa atacggaagg atctgaggtt cttatggctc ttgtatctat
     5761 cagtgaagca tcaagactaa caaacaaaag tagaacaact gttcaccgtt agatatcaaa
     5821 gggaaaactg tccatatgca cagatgaaaa cggtgtaaaa aagatagata catcagagct
     5881 tttacgagtt tttggtgcat ttaaagctgt tcaccatgaa cagatcgaca atgtaacaga
     5941 tgaacagcat gtaacaccta atagaacagg tgaaaccagt aaaacaaagc aactagaaca
     6001 tgaaattgaa cacctgagac aacttgttac agctcaacag tcacacatag acagcctgaa
     6061 acaggcgatg ctgcttatcg aatcaaagct gccgacaaca cgggagccag tgacgcctcc
     6121 cgtggggaaa aaatcatggc aattctggaa gaaatagcgc tttcagccgg caaacctgaa
     6181 gccggatctg cgattctgat aacaaactag caacaccaga acagcccgtt tgcgggcagc
     6241 aaaacccgta ccctaggtct agggcggcgg atttgtccta ctcaggagag cgttcaccga
     6301 caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg
     6361 cctctagcac gcgtaccatg ggatcccccg ggctgcagga attcgatatc
//
