LOCUS       sequence-373756-        5199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MfnG in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5199)
  AUTHORS   Wu KL, Moore JA, Miller MD, Chen Y, Lee C, Xu W, Peng Z, Duan Q, 
            Phillips GN Jr, Uribe RA, Xiao H
  TITLE     Expanding the eukaryotic genetic code with a biosynthesized 21st 
            amino acid.
  JOURNAL   Protein Sci. 2022 Oct;31(10):e4443. doi: 10.1002/pro.4443.
  PUBMED    36173166
REFERENCE   2  (bases 1 to 5199)
  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"; journalName: "Protein 
            Sci."; date: "2022-10"; volume: "31(10):e4443. doi"; pages: " 
            10.1002/pro.4443"
COMMENT     Sequence Label: pBad-MfnG
FEATURES             Location/Qualifiers
     source          1..5199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     72..91
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     325..342
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    357..497
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(423..442)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     583..605
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(761..1639)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     1779..1798
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1883..1905
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3265..3282)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3435..3521
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3613..3640
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3659..3750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3751..3819,3820..4611)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3969..3988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4782..5199,1..171)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
       61 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      121 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      181 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      241 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      301 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
      361 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
      421 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
      481 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
      541 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
      601 agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg
      661 cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
      721 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
      781 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
      841 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
      901 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
      961 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     1021 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     1081 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     1141 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     1201 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     1261 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     1321 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     1381 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     1441 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     1501 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     1561 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     1621 cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa accaattgtc
     1681 catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg ctaaccaaac
     1741 cggtaaccct gattatttgc acggagtcac actttgctat gccatagcat ttttatccat
     1801 aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc
     1861 gtttttttgg gctagaaata attttgttta actttaagaa ggagaataca tcaactagta
     1921 cgcaagttca cgtaaaaagg gtatctagag gttgaggtga ttttatgact ccagagggga
     1981 atgtgtcatt ggtggatgag agcttgttag tcggcgttac ggatgaggat cgcgcagtcc
     2041 gttctgccca ccaattctat gagcgtttaa ttggtttgtg ggcgcctgca gtaatggagg
     2101 ctgcccatga gttgggcgtt tttgctgcct tagcagaagc gcctgctgac tcaggggaac
     2161 tggctcgccg cctggattgc gacgcccgtg ctatgcgtgt attgctggac gcattatacg
     2221 catatgacgt cattgaccgc atccatgata ctaacggctt tcgttattta cttagtgccg
     2281 aggcacgtga atgccttctt ccgggtacgc tgttttcctt agtcggcaag ttcatgcatg
     2341 acatcaacgt ggcctggcct gcctggcgca atcttgctga agtggtgcgc cacggcgcgc
     2401 gtgatacgtc cggcgccgaa tctccaaatg gcatcgcaca ggaagactat gaatctttgg
     2461 tcggagggat caatttttgg gcacctccaa tcgtgacgac cttgtctcgc aaactgcgtg
     2521 cctctggacg ttcgggggat gcaacagcct ccgtgctgga tgttggctgc ggtacgggac
     2581 tgtactcaca gctgctgtta cgcgagttcc ctcgttggac agccacaggc ttagacgtcg
     2641 aacgtatcgc gactttagct aatgctcagg cactgcgctt gggagtcgaa gagcgttttg
     2701 caactcgtgc cggggatttc tggcgcggag gctggggaac aggctacgac ttagtcttgt
     2761 ttgccaatat tttccatttg caaacgccag catcagcagt ccgtcttatg cgtcatgccg
     2821 ccgcttgtct ggcgcccgac ggtttagtgg ctgtagtaga ccaaattgta gatgccgacc
     2881 gtgaacccaa aaccccacag gatcgttttg ctttattatt tgcggcctca atgaccaaca
     2941 ctggtggcgg cgacgcatat acattccaag agtatgaaga atggtttacc gcagcgggat
     3001 tgcagcgcat tgaaactttg gacactccaa tgcatcgcat tcttttagcc cgtcgcgcca
     3061 ccgaacctag cgcagtcccg gagggacagt gataagtcga ccgatgccct tgagagcctt
     3121 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     3181 tgtcttcttt atcatgcaac tcgtaggaca ggtgccaaac ggtctccagc ttggctgttt
     3241 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     3301 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     3361 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     3421 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     3481 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     3541 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     3601 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     3661 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3721 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3781 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3841 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3901 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3961 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     4021 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4081 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4141 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4201 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4261 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4321 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4381 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4441 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4501 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4561 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4621 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4681 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4741 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4801 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4861 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4921 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4981 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5041 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5101 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5161 ctacagcgtg agctatgaga aagcgccacg cttcccgaa
//
