LOCUS       sequence-213176-        6976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tyr-tRNA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6976)
  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 6976)
  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: B7-3
FEATURES             Location/Qualifiers
     source          1..6976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..80
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        61..79
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    80..104
                     /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)."
     RBS             119..141
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1423..1440
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1537..1555)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1551..1598
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1773..1778,1779..1795,1796..1801))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1828..1846)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1914..2018
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2019..2087,2088..2879)
                     /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(2237..2256)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3050..3638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3539..3558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3792..3809
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3824..3963
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3889..3908)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4049..4071
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4065..4256)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5568..6650)
                     /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     6612..6631
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6651..6728)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6934..6953
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
       61 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
      121 tgtttaactt taagaaggag atataccatg catgcaagca gtaacttgat taaacaattg
      181 caagagcggg ggctggtagc ccaggtgacg gacgaggaag cgttagcaga gcgactggcg
      241 caaggcccga tcgcgctcta ttgcggcttc gatcctaccg ctgacagctt gcatttgggg
      301 catcttgttc cattgttatg cctgaaacgc ttccagcagg cgggccacaa gccggttgcg
      361 ctggtaggcg gcgcgacggg tctgattggc gacccgagct tcaaagctgc cgagcgtaag
      421 ctgaacaccg aagaaactgt tcaggagtgg gtggacaaaa tccgtaagca ggttgccccg
      481 ttcctcgatt tcgactgtgg agaaaactct gctatcgcgg cgaacaacta tgactggttc
      541 ggcaatatga atgtgctgac cttcctgcgc gatattggca aacacttctc cgttaaccag
      601 atgatcaaca aagaagcggt taagcagcgt ctcaaccgtg aagatcaggg gatttcgttc
      661 actgagtttt cctacaacct gttgcagggt tatgacttcg cctgtctgaa caaacagtac
      721 ggtgtggtgc tgcaaattgg tggttctgac cagtggggta acatcacttc tggtatcgac
      781 ctgacccgtc gtctgcatca gaatcaggtg tttggcctga ccgttccgct gatcactaaa
      841 gcagatggca ccaaatttgg taaaactgaa ggcggcgcag tctggttgga tccgaagaaa
      901 accagcccgt acaaattcta ccagttctgg atcaacactg cggatgccga cgtttaccgc
      961 ttcctgaagt tcttcacctt tatgagcatt gaagagatca acgccctgga agaagaagat
     1021 aaaaacagcg gtaaagcacc gcgcgcccag tatgtactgg cggagcaggt gactcgtctg
     1081 gttcacggtg aagaaggttt acaggcggca aaacgtatta ccgaatgcct gttcagcggt
     1141 tctttgagtg cgctgagtga agcggacttc gaacagctgg cgcaggacgg cgtaccgatg
     1201 gttgagatgg aaaagggcgc agacctgatg caggcactgg tcgattctga actgcaacct
     1261 tcccgtggtc aggcacgtaa aactatcgcc tccaatgcca tcaccattaa cggtgaaaaa
     1321 cagtccgatc ctgaatactt ctttaaagaa gaagatcgtc tgtttggtcg ttttacctta
     1381 ctgcgtcgcg gtaaaaagaa ttactgtctg atttgctgga aacatcatca tcaccatcat
     1441 taaagcagcg gcatcaccat cattaaagca gaggccatat gctcgaggat ccggctgcta
     1501 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     1561 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     1621 gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
     1681 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
     1741 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
     1801 aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     1861 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     1921 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1981 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2041 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2101 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2161 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2221 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     2281 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2341 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2401 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2461 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2521 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     2581 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2641 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2701 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2761 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     2821 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2881 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2941 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3001 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3061 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3121 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3181 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3241 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3301 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3361 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3421 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3481 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3541 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3601 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3661 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3721 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3781 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     3841 tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc
     3901 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     3961 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4021 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4081 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     4141 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4201 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     4261 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     4321 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     4381 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     4441 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     4501 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     4561 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     4621 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     4681 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga cccaacgctg
     4741 cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat gttctgccaa
     4801 gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat tcttggagtg
     4861 gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc cggctccatg
     4921 caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca atccatgcca
     4981 acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc ggtccaatga
     5041 tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga tggtcgtcat
     5101 ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa
     5161 gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag acgtagccca
     5221 gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     5281 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     5341 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     5401 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     5461 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     5521 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     5581 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5641 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     5701 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     5761 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     5821 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     5881 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     5941 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     6001 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     6061 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     6121 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     6181 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     6241 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     6301 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     6361 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     6421 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     6481 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     6541 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     6601 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     6661 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     6721 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     6781 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     6841 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     6901 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     6961 aaccgcacct gtggcg
//
