LOCUS       40924_28202        4646 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLGFP_delta_2 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4646)
  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 4646)
  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 4646)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4646)
  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..4646
                     /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..2155
                     /codon_start=1
                     /gene="lgfp_delta_2"
                     /product="GFP deletion mutant"
                     /label=lgfp_delta_2
                     /protein_id="BAQ25563.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTL"
     gene            1796..2155
                     /gene="lgfp_delta_2"
                     /label=lgfp_delta_2
     terminator      2366..2452
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2544..2571
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2591..2682
                     /label=AmpR promoter
     CDS             2683..3540
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3573..3667
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3755..4343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4507..4593)
                     /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 tataggctag
     2161 cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc
     2221 agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac
     2281 cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat
     2341 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     2401 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     2461 agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     2521 aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     2581 acaaactctt tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2641 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2701 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2761 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2821 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2881 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     2941 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3001 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3061 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3121 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3181 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctacagc aatggcaaca
     3241 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3301 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3361 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3421 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3481 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3541 taactgtcag accaagttta cgagctcgct tggactcctg ttgatagatc cagtaatgac
     3601 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
     3661 tgagaatcca agcactagtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3721 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     3781 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3841 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3901 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4141 gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4201 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4261 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4321 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4381 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4441 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgcct aggtctaggg
     4501 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     4561 ccagtctttc gactgagcct ttcgttttat ttgatgcctc tagcacgcgt accatgggat
     4621 cccccgggct gcaggaattc gatatc
//
