LOCUS       sequence-213203-        7748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gly-tRNA synthetase B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7748)
  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 7748)
  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: B8-1
FEATURES             Location/Qualifiers
     source          1..7748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2366..2383
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2457..2475)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2471..2518
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2693..2698,2699..2715,2716..2721))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2748..2766)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2834..2938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2939..3007,3008..3799)
                     /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(3157..3176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3970..4558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4459..4478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4712..4729
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4744..4883
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4809..4828)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4969..4991
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4985..5176)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6488..7570)
                     /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     7532..7551
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7571..7648)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgagagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatgca
      301 ttctgagaaa acttttctgg tggaaatcgg cactgaagag ctgccaccaa aagcactgcg
      361 cagcctggct gagtcctttg ctgcgaactt tactgcggag ctggataacg ctggcctcgc
      421 acacggcacc gttcaatggt ttgctgctcc gcgtcgtctg gcgctgaaag tagctaacct
      481 ggcggaagcg caaccggatc gtgaaatcga aaaacgcggc ccggcgattg cccaggcgtt
      541 cgacgctgaa ggcaaaccga gcaaagcggc agaaggttgg gcgcgtggtt gcggtattac
      601 cgttgaccag gctgagcgtc tgactaccga taaaggcgaa tggctgctgt atcgcgccca
      661 tgtgaagggc gaaagcaccg aagcactgct gccgaatatg gttgcgactt ctctggcgaa
      721 actgccgatc ccgaaactga tgcgttgggg cgcaagcgac gtgcacttcg tgcgtccggt
      781 gcacaccgtg accctgctgc tgggcgacaa agtcattccg gcaaccattc tgggcattca
      841 gtccgatcgc gtgattcgcg gccaccgctt tatgggcgag ccggaattca ccatcgataa
      901 cgccgatcag tatccggaaa ttctgcgtga gcgtgggaaa gtcatcgccg attacgaaga
      961 acgtaaggcg aagattaaag ccgatgccga agaagcagcg cgtaagattg gcggtaacgc
     1021 tgacttaagc gaaagcctgc tggaagaagt ggcttcgctg gtggagtggc cggtcgttct
     1081 gaccgcaaaa ttcgaagaga aattcctcgc ggtgccggct gaagcgctgg tttacaccat
     1141 gaaaggtgac cagaaatact tcccggtgta tgcgaacgac ggcaaactgc tgccgaactt
     1201 tatcttcgtt gccaacatcg aatcgaaaga tccgcagcag attatctccg gtaacgagaa
     1261 agtcgttcgt ccgcgtctgg cggatgccga gttcttcttc aacaccgacc gtaaaaaacg
     1321 tcttgaagat aacctgccgc gcctgcaaac cgtgttgttc cagcaacagt tggggacgct
     1381 gcgcgacaaa actgaccgca tccaggcgct ggctggctgg attgctgaac agattggcgc
     1441 tgacgttaac cacgctaccc gtgcgggtct gctgtctaag tgcgacctga tgaccaacat
     1501 ggtcttcgag ttcaccgaca cccagggcgt tatggggatg cactatgcgc gtcacgatgg
     1561 cgaagcggaa gatgtcgcgg tggcgctgaa tgagcagtat cagccgcgtt ttgctggtga
     1621 tgacctgccg tccaacccag tagcttgtgc gctggcgatt gctgacaaga tggataccct
     1681 ggcgggtatc ttcggtatcg gtcagcatcc gaaaggcgac aaagacccgt ttgcgctgcg
     1741 tcgtgccgcg cttggcgtgc tgcgaattat cgttgagaag aacctcaacc ttgatctgca
     1801 aacgctgacc gaagaagcgg tgcgtctgta tggcgataag ctgactaatg ccaacgtagt
     1861 tgatgatgtt atcgacttta tgctcggtcg cttccgcgcc tggtatcagg acgaaggtta
     1921 taccgttgac accatccagg cggtactggc gcgtcgtccg actcgtccgg ctgatttcga
     1981 tgcccgtatg aaagcggtat cgcatttccg taccctggat gcagctgctg cactggcggc
     2041 ggcgaacaaa cgtgtatcta acattctggc gaaatctgac gaagtgctga gcgaccgcgt
     2101 gaatgcctct accctgaaag agccggaaga aattaaactg gcgatgcagg ttgtggtgct
     2161 acgtgacaag ctggagccgt actttacgga aggtcgttac caggatgcgc tggtcgaact
     2221 ggctgagctg cgtgaaccgg ttgatgcttt cttcgataaa gtgatggtca tggttgatga
     2281 caaagaattg cgtatcaacc gtctgaccat gctggagaaa ctgcgcgaac tgttcctgcg
     2341 cgttgcggat atttcgctgt tgcaacatca tcatcaccat cattaaagca gaggccatat
     2401 gctcgaggat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     2461 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     2521 gaaaggagga actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag
     2581 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
     2641 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
     2701 ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg
     2761 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     2821 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2881 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2941 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3001 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3061 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3121 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3181 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3241 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3301 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     3361 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3421 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3481 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3541 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     3601 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     3661 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     3721 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3781 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     3841 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3901 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3961 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4021 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4081 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4141 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4201 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4261 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4321 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4381 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4441 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4501 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4561 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4621 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4681 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4741 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     4801 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     4861 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     4921 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4981 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     5041 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     5101 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     5161 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     5221 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     5281 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     5341 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     5401 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     5461 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     5521 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     5581 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     5641 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
     5701 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
     5761 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
     5821 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
     5881 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
     5941 gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
     6001 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
     6061 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
     6121 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     6181 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     6241 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     6301 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     6361 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     6421 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     6481 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6541 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     6601 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     6661 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     6721 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
     6781 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     6841 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     6901 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     6961 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     7021 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     7081 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     7141 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     7201 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     7261 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     7321 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     7381 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     7441 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     7501 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
     7561 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
     7621 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
     7681 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
     7741 ggtgcatg
//
