LOCUS       sequence-350726-        8173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses His tagged PheRS in Escherichia coli cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8173)
  TITLE     Purified and reconstituted E. coli in-vitro translation components
REFERENCE   2  (bases 1 to 8173)
  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"
COMMENT     Sequence Label: pEcH6PheRS
FEATURES             Location/Qualifiers
     source          1..8173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      107..695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     596..615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     849..866
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    881..1020
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(946..965)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1106..1128
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1122..1313)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2771..2790
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2874..2893
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2874..2892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2981..3003
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3023..3040
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3041..3061
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(6576..6594)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6590..6637
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7003..7008,7009..7025,7026..7031))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(7058..7076)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7144..7248
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7249..7317,7318..8109)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7467..7486)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
       61 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      121 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      181 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      241 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      301 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      361 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      421 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      481 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      541 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      601 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      661 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      721 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      781 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      841 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
      901 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
      961 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     1021 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     1081 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     1141 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     1201 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     1261 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     1321 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     1381 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     1441 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     1501 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     1561 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     1621 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     1681 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     1741 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
     1801 gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg
     1861 ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg
     1921 aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac
     1981 cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc
     2041 cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg
     2101 aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta
     2161 cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa
     2221 tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg
     2281 cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
     2341 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
     2401 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
     2461 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     2521 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgacgct
     2581 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     2641 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     2701 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     2761 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     2821 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
     2881 actcactata gggagaccac aacggtttcc ctctagtgcc ggctccggag agctctttaa
     2941 ttaagcggcc gccctgcagg actcgagttc tagaaataat tttgtttaac tttaagaagg
     3001 agatatacat atgaaatctt ctcaccatca ccatcaccat gaaaacctgt acttccaatc
     3061 caatgcatca catctcgcag aactggttgc cagtgcgaag gcggccatta gccaggcgtc
     3121 agatgttgcc gcgttagata atgtgcgcgt cgaatatttg ggtaaaaaag ggcacttaac
     3181 ccttcagatg acgaccctgc gtgagctgcc gccagaagag cgtccggcag ctggtgcggt
     3241 tatcaacgaa gcgaaagagc aggttcagca ggcgctgaat gcgcgtaaag cggaactgga
     3301 aagcgctgca ctgaatgcgc gtctggcggc ggaaacgatt gatgtctctc tgccaggtcg
     3361 tcgcattgaa aacggcggtc tgcatccggt tacccgtacc atcgaccgta tcgaaagttt
     3421 cttcggtgag cttggcttta ccgtggcaac cgggccggaa atcgaagacg attatcataa
     3481 cttcgatgct ctgaacattc ctggtcacca cccggcgcgc gctgaccacg acactttctg
     3541 gtttgacact acccgcctgc tgcgtaccca gacctctggc gtacagatcc gcaccatgaa
     3601 agcccagcag ccaccgattc gtatcatcgc gcctggccgt gtttatcgta acgactacga
     3661 ccagactcac acgccgatgt tccatcagat ggaaggtctg attgttgata ccaacatcag
     3721 ctttaccaac ctgaaaggca cgctgcacga cttcctgcgt aacttctttg aggaagattt
     3781 gcagattcgc ttccgtcctt cctacttccc gtttaccgaa ccttctgcag aagtggacgt
     3841 catgggtaaa aacggtaaat ggctggaagt gctgggctgc gggatggtgc atccgaacgt
     3901 gttgcgtaac gttggcatcg acccggaagt ttactctggt ttcgccttcg ggatggggat
     3961 ggagcgtctg actatgttgc gttacggcgt caccgacctg cgttcattct tcgaaaacga
     4021 tctgcgtttc ctcaaacagt ttaaataagg caggaataga ttatgaaatt cagtgaactg
     4081 tggttacgcg aatgggtgaa cccggcgatt gatagcgatg cgctggcaaa tcaaatcact
     4141 atggcgggcc tggaagttga cggtgtagaa ccggttgccg gcagcttcca cggcgtggtc
     4201 gttggtgaag tggttgagtg tgcgcagcat ccgaacgctg acaaactgcg tgtgacaaaa
     4261 gtgaatgtcg gcggcgatcg cctgctggac atcgtctgcg gtgcgccaaa ctgccgtcag
     4321 ggcctgcgtg tagcggtagc gaccattggt gctgttctgc cgggtgattt caaaattaaa
     4381 gcggcgaaac tgcgtggcga accgtctgaa gggatgctgt gctccttctc tgaactgggc
     4441 atttctgacg atcacagcgg cattatcgaa ctgcctgcgg atgcgccgat tggcaccgat
     4501 atccgtgaat acctgaaact tgatgacaac accatcgaaa tcagcgtgac gccaaaccgt
     4561 gccgactgct taggcatcat tggtgttgcg cgtgacgttg ccgtgctgaa ccagctgccg
     4621 ctggttcaac cggaaatcgt tccggttggt gcgaccatcg acgacacgct gccgattaca
     4681 gtcgaagcgc cggaagcctg cccgcgttat cttggccgtg tggtaaaagg cattaacgtt
     4741 aaagcgccaa ctccgctgtg gatgaaagaa aaactgcgtc gttgcgggat ccgttctatc
     4801 gatgcagttg ttgacgtcac caactatgtg ctgctcgaac tgggccagcc gatgcacgct
     4861 ttcgataaag atcgcattga aggcggcatt gtggtgcgga tggcgaaaga gggcgaaacg
     4921 ctggtgctgc tcgacggtac tgaagcgaag ctgaatgctg acactctggt catcgccgac
     4981 cacaacaagg cgctggcgat gggcggcatc ttcggtggcg aacactctgg cgtgaatgac
     5041 gaaacacaaa acgtgctgct ggaatgcgcg ttctttagcc cgctgtctat caccggtcgt
     5101 gctcgtcgtc atggcctgca taccgatgcg tctcaccgtt atgagcgtgg cgttgatccg
     5161 gcactgcagc acaaagcgat ggaacgtgcg acccgtctgc tgatcgacat ctgcggtggt
     5221 gaggctggcc cggtaattga tatcaccaac gaagcaacgc tgccgaagcg tgcaaccatc
     5281 actctacgtc gtagcaaact ggatcgcctg atcggccatc atattgcgga tgagcaggta
     5341 actgacattc tgcgtcgtct cggctgcgaa gtgaccgaag gcaaagacga atggcaggca
     5401 gttgcgccga gctggcgttt cgatatggag attgaagaag atctggttga agaagtcgcg
     5461 cgtgtttacg gctacaacaa catcccggat gagccggtac aggcaagcct gattatgggt
     5521 actcaccgtg aagctgacct gtcgctcaag cgcgtgaaaa cgctgctcaa cgacaaaggc
     5581 tatcaggaag tgatcaccta cagcttcgtt gatccgaaag tgcagcagat gatccatcca
     5641 ggcgttgaag ccttactgct gccaagcccg atctctgttg aaatgtcagc aatgcgtctt
     5701 tctctgtgga ctggcctgct ggcaaccgtg gtgtacaacc agaaccgtca gcagaaccgt
     5761 gtgcgcattt tcgaaagcgg tctgcgtttc gtaccagata ctcaggcacc gttgggcatt
     5821 cgtcaggatc tgatgttagc cggtgtgatt tgcggtaacc gttacgaaga gcactggaac
     5881 ctggcaaaag agaccgttga tttctatgat ttgaaaggcg atcttgaatc cgttctcgac
     5941 ctgaccggta aactgaatga ggttgagttc cgtgcagaag cgaatccggc actgcatccg
     6001 gggcaatccg cagcgattta tctgaaaggt gaacgtattg gttttgttgg ggttgttcat
     6061 cctgaactgg aacgtaaact ggatcttaac ggtcgcactc tggtgttcga actggagtgg
     6121 aacaagctcg cagaccgcgt ggtgcctcag gcgcgcgaga tttctcgctt cccggcgaac
     6181 cgtcgtgaca tcgcggtggt ggtcgcagaa aacgttcccg cagcggatat tttatccgaa
     6241 tgtaagaaag ttggcgtaaa tcaggtagtt ggcgtaaact tatttgacgt gtaccgcggt
     6301 aagggtgttg cggaggggta taagagcctc gccataagcc tgatcctgca agataccagc
     6361 cgtacactcg aagaagagga gattgccgct accgtcgcca aatgtgtaga ggcattaaaa
     6421 gagcgattcc aggcatcatt gagggattaa taacattgga agtggataac ggatccgcga
     6481 tcgcggcgcg ccacctggtg gccggccggt accacgcgtg cgcgctgatc cggctgctaa
     6541 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     6601 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     6661 atatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg
     6721 aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga acgggtgcgc
     6781 atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg gcgatgctgt
     6841 cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     6901 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     6961 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     7021 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     7081 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     7141 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     7201 agacaataac cctgataaat gcttcaataa cattgaaaaa ggaagagtat gagtattcaa
     7261 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     7321 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     7381 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     7441 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     7501 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7561 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7621 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7681 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7741 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     7801 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7861 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     7921 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     7981 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     8041 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     8101 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     8161 ttttaattta aaa
//
