LOCUS       sequence-240275-        7793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  translational elongation factor G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7793)
  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 7793)
  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: B7-1
FEATURES             Location/Qualifiers
     source          1..7793
                     /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"
     protein_bind    293..317
                     /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             332..354
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2476..2493
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2567..2585)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2581..2628
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2803..2808,2809..2825,2826..2831))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2858..2876)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2944..3048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3049..3117,3118..3909)
                     /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(3267..3286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4080..4668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4569..4588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4822..4839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4854..4993
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4919..4938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5079..5101
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5095..5286)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6598..7680)
                     /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     7642..7661
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7681..7758)
                     /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 ggggaattgt
      301 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
      361 atgcatgctc gtacaacacc catcgcacgc taccgtaaca tcggtatcag tgcgcacatc
      421 gacgccggta aaaccactac taccgaacgt attctgttct acaccggtgt aaaccataaa
      481 atcggtgaag ttcatgacgg cgctgcaacc atggactgga tggagcagga gcaggaacgt
      541 ggtattacca tcacttccgc tgcgactact gcattctggt ctggtatggc taagcagtat
      601 gagccgcatc gcatcaacat catcgacacc ccggggcacg ttgacttcac aatcgaagta
      661 gaacgttcca tgcgtgttct cgatggtgcg gtaatggttt actgcgcagt tggtggtgtt
      721 cagccgcagt ctgaaaccgt atggcgtcag gcaaacaaat ataaagttcc gcgcattgcg
      781 ttcgttaaca aaatggaccg catgggtgcg aacttcctga aagttgttaa ccagatcaaa
      841 acccgtctgg gcgcgaaccc ggttccgctg cagctggcga ttggtgctga agaacatttc
      901 accggtgttg ttgacctggt gaaaatgaaa gctatcaact ggaacgacgc tgaccagggc
      961 gtaaccttcg aatacgaaga tatcccggca gacatggttg aactggctaa cgaatggcac
     1021 cagaacctga tcgaatccgc agctgaagct tctgaagagc tgatggaaaa atacctgggt
     1081 ggtgaagaac tgactgaagc agaaatcaaa ggtgctctgc gtcagcgcgt tctgaacaac
     1141 gaaatcatcc tggtaacctg tggttctgcg ttcaagaaca aaggtgttca ggcgatgctg
     1201 gatgcggtaa ttgattacct gccatccccg gttgacgtac ctgcgatcaa cggtatcctg
     1261 gacgacggta aagacactcc ggctgaacgt cacgcaagtg atgacgagcc gttctctgca
     1321 ctggcgttca aaatcgctac cgacccgttt gttggtaacc tgaccttctt ccgtgtttac
     1381 tccggtgtgg ttaactctgg tgataccgta ctgaactccg tgaaagctgc acgtgagcgt
     1441 ttcggtcgta tcgttcagat gcacgctaac aaacgtgaag agatcaaaga agttcgcgcg
     1501 ggcgacatcg ctgctgctat cggtctgaaa gacgtaacca ctggtgacac cctgtgtgac
     1561 ccggatgcgc cgatcattct ggaacgtatg gaattccctg agccggtaat ctccatcgca
     1621 gttgaaccga aaaccaaagc tgaccaggaa aaaatgggtc tggctctggg ccgtctggct
     1681 aaagaagacc cgtctttccg tgtatggact gacgaagaat ctaaccagac catcatcgcg
     1741 ggtatgggcg aactgcacct cgacatcatc gttgaccgta tgaagcgtga attcaacgtt
     1801 gaagcgaacg taggtaaacc gcaggttgct taccgtgaaa ctatccgcca gaaagttacc
     1861 gatgttgaag gtaaacacgc gaaacagtct ggtggtcgtg gtcagtatgg tcatgttgtt
     1921 atcgacatgt acccgctgga gccgggttca aacccgaaag gctacgagtt catcaacgac
     1981 attaaaggtg gtgtaatccc tggcgaatac atcccggccg ttgataaagg tatccaggaa
     2041 cagctgaaag caggtccgct ggcaggctac ccggtagtag acatgggtat tcgtctgcac
     2101 ttcggttctt accatgacgt tgactcctct gaactggcgt ttaaactggc tgcttctatc
     2161 gcctttaaag aaggctttaa gaaagcgaaa ccagttctgc ttgagccgat catgaaggtt
     2221 gaagtagaaa ctccggaaga gaacaccggt gacgttatcg gtgacttgag ccgtcgtcgt
     2281 ggtatgctca aaggtcagga atctgaagtt actggcgtta agatccacgc tgaagtaccg
     2341 ctgtctgaaa tgttcggata cgcaactcag ctgcgttctc tgaccaaagg tcgtgcatca
     2401 tacactatgg aattcctgaa gtatgatgaa gcgccgagta acgttgctca ggccgtaatt
     2461 gaagcccgtg gtaaacatca tcatcaccat cattaaagca gaggccatat gctcgaggat
     2521 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2581 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga
     2641 actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     2701 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     2761 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     2821 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     2881 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2941 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3001 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3061 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3121 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3181 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3241 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     3301 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3361 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3421 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3481 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3541 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     3601 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3661 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3721 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3781 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3841 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3901 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3961 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4021 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4201 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4741 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4801 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4861 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     4921 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     4981 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5041 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5101 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     5161 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     5221 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     5281 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     5341 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     5401 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     5461 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     5521 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     5581 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     5641 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     5701 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     5761 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     5821 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     5881 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     5941 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     6001 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     6061 ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     6121 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     6181 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     6241 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     6301 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     6361 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     6421 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     6481 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     6541 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     6601 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6661 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     6721 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     6781 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     6841 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     6901 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     6961 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     7021 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     7081 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     7141 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     7201 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     7261 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     7321 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     7381 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     7441 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     7501 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     7561 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     7621 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     7681 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     7741 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgc
//
