LOCUS       sequence-83944-e        5525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for in vitro transcription/translation of soybean His-tagged 
            arginyl-tRNA synthetase in the Cell Free Sciences system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5525)
  TITLE     Wheat germ in vitro translation of soybean arginyl-tRNA synthetase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5525)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pEU-Gm23
FEATURES             Location/Qualifiers
     source          1..5525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             119..136
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2612..2628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2612..2628)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2625..2647)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2636..2652
                     /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)."
     promoter        complement(join(2660..2666,2667..2684,2685..2690))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2705..2726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3014..3602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3094..3113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3773..4564,4565..4633))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIRYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4396..4415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4634..4738)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4806..4824
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4862..4884)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4984..5003
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5217..5236
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atttaggtga cactatagaa ctcacctatc tccccaacac ctaataacat tcaatcactc
       61 tttccactaa ccacctatct acatcaccaa gatatcacta gttctcgagc tcatgaaaca
      121 tcatcaccat caccactcga cctgggcgta tgttgaagta tattgtccag ctagtgttaa
      181 gaagcagttg gcgaaggagt ttgaggagtc tctccggata accgtgcctg atgaacccga
      241 tgttgtgcct tctattgatc cttgcgctgc caacaaagct ggtattaaat tcgctgatta
      301 tcaatgtaat aatgcaatgg gtctatttgc taagatgaaa gggaagcaga caggattcag
      361 gggtccccag gcagttggac aggccatact taaccatctt ccgcagtctg aaatgataga
      421 atcatgctct gtagctggtc ctgggtttgt gaatattgtc ttgtcgaaga agtggatagc
      481 gcagaggtta gagaggttgc tgattgatgg tattgataag tgggcacctc gacttcccgt
      541 gaagacagtt ctggttgatt tttcctcacc taacatagca aaggaaatgc atgttggaca
      601 cctgagatct accattattg gggacacatt agctcgcatg tttgaatttt cacgtgtgga
      661 aactcttatc cgcagaaatc atcttggtga ctgggggact cagtttggga tgctaatagc
      721 acacctcttt gacacttatc caaacccaga agattttaat gaagcagcta ttggggatct
      781 tcaggcattc tataaagcct ccaaagtgag gtttgataat gatcctgaat ttaagttgag
      841 ggcacaacag tcagtggtcc ggctccagag tggagaagaa aagtatcaca gggcatggca
      901 gcaaatttgt gatattagtc aggctgaatt tgaaaaggtc tatcaacgtc ttggagttcg
      961 attggaagca aggggagaga gctactataa tgatttaatc cctcaaactt tggagagatt
     1021 ggataattta ggactgattg aagaagatga tggtgctcgt gtgatatttg ttgagggtgt
     1081 aaatatacca attattgctg tgaaaagaga tggtggctac aactattctt caactgatct
     1141 agcagcactt tggaatcgtc taaatgtgga aaatgttgag tggaatatat acgttacaga
     1201 tgttgggcag tggcaacact ttgatatggt ttttaaggcc tttaggcgtg caggatggct
     1261 gccaaatcat gaaaatgagt tccccaaatg tactcatatt ggttttggtc ttgttcttgg
     1321 ggaagatgga aaacgatttc ggacacgcag cagtgaaact gttagattgg ttgagttgct
     1381 tgatgaagcc aaaaggcgtt gtaaagcttc cctcctcgag cgtgatgctg ttaaagattg
     1441 gtctgaggaa gagatcgaaa agacagcaga agcagttggt tacggagctg ttaagtatgc
     1501 tgacttgagg atcaacagat taacgaatta cacattcagc tttgatcaga tgcttaatga
     1561 taagggaaat actgctgttt atttgcagta tgcgcatgct aggatctgtt ctatcatgag
     1621 gaaatctggt aaagatatgg aagaaataaa gaaaaatggg aatatagtgt cgaatcatga
     1681 agatgaacgt gcattggggc ttcatttgat acaatttcct gaggtttttg aggagtcgtt
     1741 gaccaatttg ttgcccaatg tgctgtgtga atacctctat aatttgacag aaatctttac
     1801 aaaaaaattt tatactaatt gccaggttgt ggggtcgcct gaggagacta gtagactctt
     1861 gctatgtgaa gcaacagtga ctgtgatgag acaatgcttt tatctccttg gaattgaacc
     1921 tgtatacagg ctttgaggat ccatatatag ggcccgggtt ataattacct caggtcgacg
     1981 tcccatggtt ttgtatagaa tttacggcta gcgccggatg cgacgccggt cgcgtcttat
     2041 ccggccttcc tatatcaggc ggtgtttaag acgccgccgc ttcgcccaaa tccttatgcc
     2101 ggttcgacga ctggacaaaa tactgtttat cttcccagcg caggcaggtt aatgtaccac
     2161 cccagcagca gccggtatcc agcgcgtata taccttccgg cgtacctttg ccctccagcg
     2221 atgcccagtg accaaaggcg atgctgtatt cttcagcgac agggccagga atcgcaaacc
     2281 acggtttcag tggggcaggg gcctcttccg gcgattctta ctagctagta tgcataggtg
     2341 ctgaaatata aagtttgtgt ttctaaaaca cacgtggtac gtacgataac gtacagtgtt
     2401 tttccctcca cttaaatcga agggtagtgt cttggagcgc gcggagtaaa catatatggt
     2461 tcatatatgt ccgtaggcac gtaaaaaaag cgagggattc gaattccccc ggaacccccg
     2521 gttggggccc acgcctcgat cgagcaaaaa aaaaaaaaaa gaaaaaaaaa aaaaaaaaaa
     2581 gctttcccgc ggccagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     2641 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     2701 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2761 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2821 gtattgggcg ctcttccgct tcctcactca ctgactcgct gcgctcggtc gctcggctgc
     2881 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2941 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3001 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3061 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3121 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3181 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3241 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3301 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3361 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3421 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3481 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3541 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3601 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3661 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3721 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3781 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3841 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3901 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3961 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4021 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4081 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4141 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4201 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4261 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4321 gtgagtactc aaccaagtca ttctgagaat agcgtatgcg gcgaccgagt tgctcttgcc
     4381 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4441 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4501 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4561 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4621 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4681 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4741 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     4801 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     4861 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4921 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     4981 atgcggcatc agagcagatt gtactgagag tgcaccattc gacgctctcc cttatgcgac
     5041 tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg
     5101 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     5161 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     5221 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     5281 cgtccggcgt agaggatctg gctagcgatg accctgctga ttggttcgct gaccatttcc
     5341 gggtgcggga cggcgttacc agaaactcag aaggttcgtc caaccaaacc gactctggcg
     5401 gcagtttacg agagagatga tagggtctgc ttcagtaagc cagatgctac acaattaggc
     5461 ttgtacatac tgtcgttaga acgcggctac aattaataca taaccttatg tatcatacac
     5521 atacg
//
