LOCUS       sequence-78029-e        7693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a plant arginyl-tRNA synthetase in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7693)
  AUTHORS   Hogg J, Schiefermayr E, Schiltz E, Igloi GL
  TITLE     Expression and properties of arginyl-tRNA synthetase from jack bean 
            (Canavalia ensiformis).
  JOURNAL   Protein Expr Purif. 2008 Oct;61(2):163-7. doi: 
            10.1016/j.pep.2008.05.008. Epub 2008 May 23.
  PUBMED    18562210
REFERENCE   2  (bases 1 to 7693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.pep.2008.05.008"; journalName: "Protein Expr Purif"; 
            date: "2008-10"; volume: "61"; issue: "2"; pages: "163-7"
COMMENT     Sequence Label: pET-Ce25
FEATURES             Location/Qualifiers
     source          1..7693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2012..2026)
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             complement(2042..2086)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(2093..2110)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(2120..2137)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2159..2485)
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     RBS             2493..2515
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2530..2554
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     complement(2554..2573)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2555..2573)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2661..2680)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2886..2963
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2964..4046
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2983..3002)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4855..5046
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5040..5062)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5148..5290
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5203..5222
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5305..5322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5476..6064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5556..5575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6235..7026,7027..7095))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6858..6877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7096..7200)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7227..7682)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7364..7385)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7576..7595
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgatatca gccatggtca tagcttgtat acagggtcaa
      241 tcccaaggag ataaaagcac tttcgcatca cagttgccgt tgcttcacat agtaagagtc
      301 tactagtttg ctcaggcgac ccaatgacct gacaattagt ataaaacttt ttggtaaaga
      361 tttctgtcaa attatagagg tattcacaca acacattggg caataaattg gccaatgact
      421 ccacaaagac ctcaggaaat tgtatcaagt gaagccccaa tgcacgctca tcttcatgat
      481 tcaatactat attcccattt ttctttaatt cttctatatc tttaccagat ttcctgataa
      541 tggaacagat cctagcatgt gcatatagca aataaacagc agtgttcccc ttatcattaa
      601 gcatctgatc gaagttgaat gtgtagtttg ttaatctgtt gattttcaag tcagcatact
      661 taacagcccc ataaccaatt gcctctgatg ttttttcaat ctcttcttca gaccaatctt
      721 tagcagcatc acgttcaagc agggcagatt tacaacgcct tttggcttca tctagtagat
      781 caaccaatcg aacagtttcg ctgctgcgac tccgaaatcg ttttccatct tccccaagaa
      841 caagaccaaa accaatatgg gtacattttg ggaagtcatt ttcatccttt ggcagccatc
      901 ctgcacgcct gaaggcctta aaaagcatat caaagtgctg ccactgccca acatctgtaa
      961 cataaatgtt ccactcaagt ttttccacat ttaggcgata ccaaagtgac gctagatcag
     1021 atgaaaaata gttgtagcca ccatctcttt tcacagcaat aattggtatg tttacacctt
     1081 caacaaatat cacacgagca ccatcactgt cttcaatcag tcctaattta tccaacttct
     1141 ccaaagttgg agggattaaa tcattaaaga aactctctcc cattgcctct actcgaactc
     1201 caagccgttt atagaccttt tcaaattcag ccttgctaac atcacaaatt cgctcccatg
     1261 ctttgcgata cttcttgtcc ccactctgga gaaggactac tgactgttgt gccctcagct
     1321 taaattcagg atcattatca aacctcactt ttgaggcttt atagaatgcc tgaagatccc
     1381 caatagctgt ttcattaaca tcttctgggt tcggatacat gtcaaagagg tgtgctatta
     1441 gcataccaaa ctgtgttccc cagtcaccga gatgattccg gcggatgaga gtttctacac
     1501 atgcaaattc aagcatacga gctaatgtat ccccaataat ggtagacctc agatgaccaa
     1561 catgcatttc ctttgctatg ttaggtgagg aaaactcaac cataactacc ttcaccggga
     1621 gtcgaggtgc ccaggtatca ataccatcaa tcagcatcat ctctaacctc tgcgctatcc
     1681 acttctttga taagacaata ttcacaaatc caggaccagc gatagagcat gattctatca
     1741 tttcagactg tggaagattg gcaacaatag cctgtccaag tgcaggggga cccctgaatc
     1801 ctgtctgctt tccttttatc ttagcccaca aactcattgc attattacat tgataatcag
     1861 caaatttcac accagatttc gtatcgcaag catcaactaa aggcgctaca tccggttcat
     1921 caggcacagt tgccctcaga gacacctcaa acagctttgc cagctgtttt ttaacactgg
     1981 ccggacaatc cagttcaata aaaaccatgg ccttgtcgtc gtcgtcggta cccagatctg
     2041 ggctgtccat gtgctggcgt tcgaatttag cagcagcggt ttctttcata ccagaaccgc
     2101 gtggcaccag accagaagaa tgatgatgat gatggtgcat atggccagaa ccagaaccgg
     2161 ccaggttagc gtcgaggaac tctttcaact gacctttaga cagtgcaccc actttggttg
     2221 ccgccacttc accgtttttg aacagcagca gagtcgggat accacggatg ccatatttcg
     2281 gcgcagtgcc agggttttga tcgatgttca gttttgcaac ggtcagtttg ccctgatatt
     2341 cgtcagcgat ttcatccaga atcggggcga tcattttgca cggaccgcac cactctgccc
     2401 agaaatcgac gaggatcgcc ccgtccgctt tgagtacatc cgtgtcaaaa ctgtcgtcag
     2461 tcaggtgaat aattttatcg ctcatatgta tatctccttc ttaaagttaa acaaaattat
     2521 ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc
     2581 gggatcgaga tcgatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     2641 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     2701 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     2761 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     2821 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga
     2881 tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     2941 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     3001 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     3061 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     3121 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     3181 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     3241 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     3301 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     3361 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     3421 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     3481 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     3541 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     3601 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     3661 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     3721 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggacatctcg gtagtgggat
     3781 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     3841 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     3901 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     3961 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     4021 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     4081 taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     4141 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     4201 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     4261 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     4321 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     4381 gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     4441 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     4501 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     4561 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
     4621 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
     4681 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
     4741 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
     4801 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
     4861 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
     4921 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
     4981 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     5041 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     5101 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     5161 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     5221 tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     5281 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5341 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5401 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5461 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5521 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5581 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5641 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5701 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5761 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5821 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5881 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5941 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6001 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6061 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6121 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6181 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6241 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6301 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6361 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6421 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6481 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6541 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6601 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6661 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6721 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6781 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6841 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6901 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6961 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7021 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7081 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7141 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7201 cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt taaaattcgc
     7261 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     7321 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     7381 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     7441 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     7501 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     7561 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     7621 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     7681 gtcccattcg cca
//
