LOCUS       sequence-213238-        8066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phe-tRNA synthetase B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8066)
  AUTHORS   Villarreal F, Contreras-Llano LE, Chavez M, Ding Y, Fan J, Pan T, 
            Tan C
  TITLE     Synthetic microbial consortia enable rapid assembly of pure 
            translation machinery.
  JOURNAL   Nat Chem Biol. 2018 Jan;14(1):29-35. doi: 10.1038/nchembio.2514. 
            Epub 2017 Nov 13.
  PUBMED    29131146
REFERENCE   2  (bases 1 to 8066)
  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: 
            "10.1038/nchembio.2514"; journalName: "Nat Chem Biol"; date: 
            "2018-01"; volume: "14"; issue: "1"; pages: "29-35"
COMMENT     Sequence Label: B6-1
FEATURES             Location/Qualifiers
     source          1..8066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     171..190
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     274..293
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        274..292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             332..354
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2749..2766
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2840..2858)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2854..2901
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3076..3081,3082..3098,3099..3104))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3131..3149)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3217..3321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3322..3390,3391..4182)
                     /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(3540..3559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4353..4941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4842..4861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5095..5112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5127..5266
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5192..5211)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5352..5374
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5368..5559)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6871..7953)
                     /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     7915..7934
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7954..8031)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
       61 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
      121 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
      181 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
      241 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattga
      301 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
      361 atgcataaat tcagtgaact gtggttacgc gaatgggtga acccggcgat tgatagcgat
      421 gcgctggcaa atcaaatcac tatggcgggc ctggaagttg acggtgtaga accggttgcc
      481 ggcagcttcc acggcgtggt cgttggtgaa gtggttgagt gtgcgcagca tccgaacgct
      541 gacaaactgc gtgtgacaaa agtgaatgtc ggcggcgatc gcctgctgga catcgtctgc
      601 ggtgcgccaa actgccgtca gggcctgcgt gtagcggtag cgaccattgg tgctgttctg
      661 ccgggtgatt tcaaaattaa agcggcgaaa ctgcgtggcg aaccgtctga agggatgctg
      721 tgctccttct ctgaactggg catttctgac gatcacagcg gcattatcga actgcctgcg
      781 gatgcgccga ttggcaccga tatccgtgaa tacctgaaac ttgatgacaa caccatcgaa
      841 atcagcgtga cgccaaaccg tgccgactgc ttaggcatca ttggtgttgc gcgtgacgtt
      901 gccgtgctga accagctgcc gctggttcaa ccggaaatcg ttccggttgg tgcgaccatc
      961 gacgacacgc tgccgattac agtcgaagcg ccggaagcct gcccgcgtta tcttggccgt
     1021 gtggtaaaag gcattaacgt taaagcgcca actccgctgt ggatgaaaga aaaactgcgt
     1081 cgttgcggga tccgttctat cgatgcagtt gttgacgtca ccaactatgt gctgctcgaa
     1141 ctgggccagc cgatgcacgc tttcgataaa gatcgcattg aaggcggcat tgtggtgcgg
     1201 atggcgaaag agggcgaaac gctggtgctg ctcgacggta ctgaagcgaa gctgaatgct
     1261 gacactctgg tcatcgccga ccacaacaag gcgctggcga tgggcggcat cttcggtggc
     1321 gaacactctg gcgtgaatga cgaaacacaa aacgtgctgc tggaatgcgc gttctttagc
     1381 ccgctgtcta tcaccggtcg tgctcgtcgt catggcctgc ataccgatgc gtctcaccgt
     1441 tatgagcgtg gcgttgatcc ggcactgcag cacaaagcga tggaacgtgc gacccgtctg
     1501 ctgatcgaca tctgcggtgg tgaggctggc ccggtaattg atatcaccaa cgaagcaacg
     1561 ctgccgaagc gtgcaaccat cactctacgt cgtagcaaac tggatcgcct gatcggccat
     1621 catattgcgg atgagcaggt aactgacatt ctgcgtcgtc tcggctgcga agtgaccgaa
     1681 ggcaaagacg aatggcaggc agttgcgccg agctggcgtt tcgatatgga gattgaagaa
     1741 gatctggttg aagaagtcgc gcgtgtttac ggctacaaca acatcccgga tgagccggta
     1801 caggcaagcc tgattatggg tactcaccgt gaagctgacc tgtcgctcaa gcgcgtgaaa
     1861 acgctgctca acgacaaagg ctatcaggaa gtgatcacct acagcttcgt tgatccgaaa
     1921 gtgcagcaga tgatccatcc aggcgttgaa gccttactgc tgccaagccc gatctctgtt
     1981 gaaatgtcag caatgcgtct ttctctgtgg actggcctgc tggcaaccgt ggtgtacaac
     2041 cagaaccgtc agcagaaccg tgtgcgcatt ttcgaaagcg gtctgcgttt cgtaccagat
     2101 actcaggcac cgttgggcat tcgtcaggat ctgatgttag ccggtgtgat ttgcggtaac
     2161 cgttacgaag agcactggaa cctggcaaaa gagaccgttg atttctatga tttgaaaggc
     2221 gatcttgaat ccgttctcga cctgaccggt aaactgaatg aggttgagtt ccgtgcagaa
     2281 gcgaatccgg cactgcatcc ggggcaatcc gcagcgattt atctgaaagg tgaacgtatt
     2341 ggttttgttg gggttgttca tcctgaactg gaacgtaaac tggatcttaa cggtcgcact
     2401 ctggtgttcg aactggagtg gaacaagctc gcagaccgcg tggtgcctca ggcgcgcgag
     2461 atttctcgct tcccggcgaa ccgtcgtgac atcgcggtgg tggtcgcaga aaacgttccc
     2521 gcagcggata ttttatccga atgtaagaaa gttggcgtaa atcaggtagt tggcgtaaac
     2581 ttatttgacg tgtaccgcgg taagggtgtt gcggaggggt ataagagcct cgccataagc
     2641 ctgatcctgc aagataccag ccgtacactc gaagaagagg agattgccgc taccgtcgcc
     2701 aaatgtgtag aggcattaaa agagcgattc caggcatcat tgagggatca tcatcatcac
     2761 catcattaaa gcagaggcca tatgctcgag gatccggctg ctaacaaagc ccgaaaggaa
     2821 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     2881 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc cgcaagaggc
     2941 ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga
     3001 tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact
     3061 gtgataaact accgcattaa agcttatcga tgataagctg tcaaacatga gaattcttga
     3121 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3181 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3241 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3301 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3361 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3421 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3481 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3541 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     3601 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3661 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3721 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3781 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3841 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     3901 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3961 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4021 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4081 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4141 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4201 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4261 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4321 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4381 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4441 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     4501 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4561 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4621 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4681 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4741 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4801 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4861 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4921 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4981 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5041 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5101 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     5161 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     5221 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     5281 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5341 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5401 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     5461 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     5521 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     5581 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     5641 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     5701 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     5761 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     5821 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     5881 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     5941 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     6001 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt
     6061 gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt
     6121 cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag
     6181 gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg
     6241 gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc
     6301 gccgaggcgg cataaatcgc cgtgacgatc agcggtccaa tgatcgaagt taggctggta
     6361 agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc
     6421 atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag
     6481 gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg
     6541 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     6601 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     6661 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     6721 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     6781 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     6841 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     6901 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6961 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     7021 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     7081 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     7141 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     7201 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     7261 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     7321 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     7381 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     7441 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     7501 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     7561 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     7621 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     7681 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     7741 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     7801 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     7861 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     7921 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     7981 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     8041 gacgctctcc cttatgcgac tcctgc
//
