LOCUS       40924_25276        6793 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pH_delta_NG DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6793)
  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 6793)
  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 6793)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6793)
  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..6793
                     /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..2332
                     /codon_start=1
                     /gene="hgfp_delta_1"
                     /product="GFP deletion mutant"
                     /label=hgfp_delta_1
                     /protein_id="BAQ25589.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQL"
     gene            1796..2332
                     /gene="hgfp_delta_1"
                     /label=hgfp_delta_1
     terminator      2543..2629
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2721..2748
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        2768..2859
                     /label=AmpR promoter
     CDS             2860..3717
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      3750..3844
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3932..4520
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4744..5457)
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     promoter        complement(5482..5555)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     CDS             5859..5924
                     /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"
     CDS             6140..6760
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline repressor TetR"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
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 ccggtatgag
     1801 caaaggcgaa gaactgttta ccggcgtggt gccgattctg gtggaactgg atggcgatgt
     1861 gaacggccag aaatttagcg tgagcggcga aggcgaaggc gatgcgacct atggcaaact
     1921 gaccctgaaa tttatttgca ccaccggcaa actgccggtg ccgtggccga ccctggtgac
     1981 cacctttggc tatggcgtgc agtgctttgc gcgctatccg gatcatatga aacagcatga
     2041 tttttttaaa agcgcgatgc cggaaggcta tgtgcaggaa cgcaccattt tttataaaga
     2101 tgatggcaac tataaaaccc gcgcggaagt gaaatttgaa ggcgataccc tggtgaaccg
     2161 cattgaactg aaaggcattg attttaaaga agatggcaac attctgggcc ataaaatgga
     2221 atataactat aacagccata acgtgtatat tatggcggat aaaccgaaaa acggcattaa
     2281 agtgaacttt aaaattcgcc ataacattaa agatggcagc gtgcagctgt aagctagctt
     2341 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
     2401 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     2461 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
     2521 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     2581 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     2641 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     2701 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     2761 aactcttttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2821 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2881 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2941 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     3001 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     3061 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     3121 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3181 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3241 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3301 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3361 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctacagcaat ggcaacaacg
     3421 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3481 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3541 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3601 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3661 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3721 ctgtcagacc aagtttacga gctcgcttgg actcctgttg atagatccag taatgacctc
     3781 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
     3841 gaatccaagc actagtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3901 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3961 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     4021 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4081 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4141 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4201 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4261 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4321 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4381 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4441 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4501 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4561 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4621 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgcctagc ttttatttgt
     4681 atagttcatc catgccatgt gtaatcccag caggggctgc aggaattcga tatcaagctt
     4741 ttatttgtat agttcatcca tgccatgtgt aatcccagca gctgttacaa actcaagaag
     4801 gatcatgtga tctctctttt cgttgggatc tttggaaagg gcagattgtg tggacaggta
     4861 atggttgtct ggtaaaagga cagggccatc gccaattgga gtattttgtt gataatggtc
     4921 tgctaattga acgcttccat ctttaatgtt gtgtctaatt ttgaagttaa ctttgattcc
     4981 attctttggt ttgtctgcca tgatgtatac attatgtgag ttatagttgt attccatttt
     5041 gtgtccaaga atgtttccat cttctttaaa atcaatacct tttaactcga ttctattaac
     5101 aagggtatca ccttcaaact tgacttcagc acgtgtcttg tagttcccgt catctttgta
     5161 aaatatagtt ctttcctgta cataaccttc gggcatggca ctcttgaaaa agtcatgctg
     5221 tttcatatga tctgggtatc tcgcaaagca ttgaacacca taaccgaaag tagtgacaag
     5281 tgttggccat ggaacaggta gcttcccagt agtgcaaata aatttaaggg taagttttcc
     5341 gtatgttgca tcaccttcac cctctccact gacagagaat ttttgcccat taacatcgcc
     5401 atctaattca acaagaattg ggacaactcc agtgaaaagt tcttctcctt tactcatgac
     5461 gtccatggtc tgtttcctcc tggtcagtgc gtcctgctga tgtgctcagt atctctatca
     5521 ctgataggga tgtcaatctc tatcactgat agggagcatg cacagataac catctgcggt
     5581 gataaattat ctctggcggt gttgacataa ataccactgg cggttataat gagcacatca
     5641 gcaggacgca ctgaccacca tgaaggtgac gctcttaaaa attaagccct gaagaagggc
     5701 agcattcaaa gcagaaggct ttggggtgtg tgatacgaaa cgaagcattg gccgtaagtg
     5761 cgattccgga ttagctgcca atgtgccaat cgcggggggt tttcgttcag gactacaact
     5821 gccacacacc accaaagcta actgacagga gaatccagat ggatgcacaa acacgccgcc
     5881 gcgaacgtcg cgcagagaaa caggctcaat ggaaagcagc aaatcccctg ttggttgggg
     5941 taagcgcaaa accagttccg aaagattttt ttaactataa acgctgatgg aagcgtttat
     6001 gcggaagagg taaagccctt cccgagtaac aaaaaaacaa cagcataaat aaccccgctc
     6061 ttacacattc cagccctgaa aaagggcatc aaattaaacc acacctatgg tgtatgcaaa
     6121 ggacggccgg atggacgtca tgtctagatt agataaaagt aaagtgatta acagcgcatt
     6181 agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct
     6241 aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc
     6301 cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag
     6361 ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg
     6421 cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga
     6481 aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact
     6541 cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc
     6601 taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac gacaagctat
     6661 cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat
     6721 atgcggatta gaaaaacaac ttaaatgtga aagtgggtct taaggatccc ccgggctgca
     6781 ggaattcgat atc
//
