LOCUS       sequence-331409-        8649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses machinery necessary for selenocysteine incorporation at 
            UAG codons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8649)
  AUTHORS   Mukai T, Sevostyanova A, Suzuki T, Fu X, Soll D
  TITLE     A Facile Method for Producing Selenocysteine-Containing Proteins.
  JOURNAL   Angew Chem Int Ed Engl. 2018 Jun 11;57(24):7215-7219. doi: 
            10.1002/anie.201713215. Epub 2018 May 9.
  PUBMED    29631320
REFERENCE   2  (bases 1 to 8649)
  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: 
            "10.1002/anie.201713215"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2018-06-11- 11"; volume: "57"; issue: "24"; pages: 
            "7215-7219"
COMMENT     Sequence Label: pSecUAG-Evol2
FEATURES             Location/Qualifiers
     source          1..8649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..285)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(59..78)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             312..1190
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(1346..1368)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1454..1594
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1509..1528
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2294..2313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2579..2670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2671..3486
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2671..3486
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2744..2763)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4741..4788
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     terminator      complement(8214..8300)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     8453..8470
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
ORIGIN
        1 atggagaaac agtagagagt tgcgataaaa agcgtcaggt aggatccgct aatcttatgg
       61 ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa ataatcaatg tggacttttc
      121 tgccgtgatt atagacactt ttgttacgcg tttttgtcat ggctttggtc ccgctttgtt
      181 acagaatgct tttaataagc ggggttaccg gtttggttag cgagaagagc cagtaaaaga
      241 cgcagtgacg gcaatgtctg atgcaatatg gacaattggt ttcttctctg aatggcggga
      301 gtatgaaaag tatggctgaa gcgcaaaatg atcccctgct gccgggatac tcgtttaatg
      361 cccatctggt ggcgggttta acgccgattg aggccaacgg ttatctcgat ttttttatcg
      421 accgaccgct gggaatgaaa ggttatattc tcaatctcac cattcgcggt cagggggtgg
      481 tgaaaaatca gggacgagaa tttgtttgcc gaccgggtga tattttgctg ttcccgccag
      541 gagagattca tcactacggt cgtcatccgg aggctcgcga atggtatcac cagtgggttt
      601 actttcgtcc gcgcgcctac tggcatgaat ggcttaactg gccgtcaata tttgccaata
      661 cggggttctt tcgcccggat gaagcgcacc agccgcattt cagcgacctg tttgggcaaa
      721 tcattaacgc cgggcaaggg gaagggcgct attcggagct gctggcgata aatctgcttg
      781 agcaattgtt actgcggcgc atggaagcga ttaacgagtc gctccatcca ccgatggata
      841 atcgggtacg cgaggcttgt cagtacatca gcgatcacct ggcagacagc aattttgata
      901 tcgccagcgt cgcacagcat gtttgcttgt cgccgtcgcg tctgtcacat cttttccgcc
      961 agcagttagg gattagcgtc ttaagctggc gcgaggacca acgtatcagc caggcgaagc
     1021 tgcttttgag caccacccgg atgcctatcg ccaccgtcgg tcgcaatgtt ggttttgacg
     1081 atcaactcta tttctcgcgg gtatttaaaa aatgcaccgg ggccagcccg agcgagttcc
     1141 gtgccggttg tgaagaaaaa gtgaatgatg tagccgtcaa gttgtcataa ttggtaacga
     1201 atcagacaat tgacggcttg acggagtagc atagggtttg cagaatccct gcttcgtcca
     1261 tttgacaggc acattatgca tgccgcttcg ccttcgcgcg cgaattgatc tgctgcctcg
     1321 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     1381 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     1441 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     1501 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     1561 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     1621 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1681 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1741 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccggccggc
     1801 cgaaatcggc cggcccggaa tagctgttcg ttgacttgat agaccgattg attcatcatc
     1861 tcataaataa agaaaaacca ccgctaccaa cggtggtttt ctcaaggttc gctgagctac
     1921 caactctttg aaccaaggta agtgggttgg aggaccgcac tcaccaaaat ctgttctttc
     1981 agtttagcct taacaggtgc ataacttcaa gacaaagtcc tctaaatcag ttaccaatgg
     2041 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2101 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2161 cgacctacac cgaactgaga taccaacagc gtgagctatg agaaagcgcc acgcttcccg
     2221 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2281 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2341 ggcttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcct
     2401 gcggcgttgg cttcttccgg tgctttgctt tttgctcaca tgttctttcc ggctttatcc
     2461 cctgattctg tggataaccg tattaccgct tttgagtgag ctgacaccgc tcgccgcagt
     2521 cgaacgaccg agcgtagcga gtcagtgagc gaggaagcgg aagagcgctg catgcctatt
     2581 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2641 atgcttcaat aatattgaaa aaggaagagt atgagccata ttcaacggga aacgtcttgc
     2701 tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc
     2761 gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca
     2821 gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc
     2881 agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact
     2941 cctgatgatg catggttact caccactgcg atccccggga aaacagcatt ccaggtatta
     3001 gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg
     3061 ttgcattcga ttcctgtttg taattgtcct tttaacagcg accgcgtatt tcgtctcgct
     3121 caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt
     3181 aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg
     3241 gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa
     3301 ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc
     3361 atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa
     3421 tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt
     3481 ttctaagaat taataactgc caggaggtgc aagatgattt tttccgtcga caaagtgcgg
     3541 gccgactttc cggtgctttc gcgtgaggta aacggtttgc cgctggctta tctcgacagc
     3601 gccgccagtg cgcagaaacc gagccaggtg attgacgccg aggccgagtt ttatcgtcat
     3661 ggctacgcgg cggtgcatcg tggtattcat accttaagcg cccaggcgac cgagaaaatg
     3721 gagaacgtgc gcaagcgggc atcgctgttt attaatgccc gttcggcgga agagctggtg
     3781 ttcgtccgcg gcacgacgga agggatcaat ctggtcgcca atagctgggg caacagcaac
     3841 gtgcgggcgg gcgataacat catcatcagt cagatggagc accacgctaa cattgttccc
     3901 tggcagatgc tttgcgcacg cgttggcgca gagctgcgtg tgatcccgct caatcccgat
     3961 ggtacgttgc aactggagac gctgcctacg ctgtttgatg agaaaactcg cctgctggca
     4021 attactcatg tctccaacgt gcttggcaca gaaaatccac tggcggaaat gatcacgctt
     4081 gcgcaccagc atggcgcaaa agtgctggtg gatggcgctc aggcggtgat gcatcatccg
     4141 gtggatgttc aggcgctgga ttgcgacttt tacgtgttct ccgggcataa actgtatggc
     4201 cccaccggaa ttggcattct ttatgtgaaa gaagccttgt tgcaggagat gccgccgtgg
     4261 gaagggggcg gttctatgat cgccaccgtc agcctgagtg aaggcactac ctggaccaaa
     4321 gcaccatggc ggtttgaagc cggtacaccc aataccgggg gcatcattgg tcttggcgcg
     4381 gcgctggagt atgtttcggc gctggggctt aataacatag ccgagtatga acagaatctg
     4441 atgcattatg cgctatcaca gctggaatct gtaccggatc tcactctcta tggcccacaa
     4501 aacaggcttg gcgttattgc ttttaatctc ggtaaacacc acgcctatga tgttggcagt
     4561 tttctcgata attacggcat tgctgtgcgt accggacatc acgccgcaat gccattgatg
     4621 gcctattaca acgtccctgc gatgtgtcgg gcgtcgctgg ccatgtataa cacccatgaa
     4681 gaagtggatc gtctggtgac cggcctgcaa cgtattcacc gtttgctggg ataatcacgt
     4741 gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac
     4801 taaaccgtga cgaactcgtc tcacgcgcca gccatcgcca gttctgctag tcagcccgaa
     4861 tcatgtccca ctgccgacga ttcactgcca gattcactgc cagattcact gccacagccc
     4921 agccagcaac aagcgcgccg tctaccgcaa gtggaacagc tgctgcagca accctttctc
     4981 accggtttta tcgaggcgct gagccgccct tttgtgacct cggcggtgcg cgatgtcctg
     5041 agcgaattgc gccagagcga ggcatttcgc cagcatgggg ttgcccccga gcaaatcgag
     5101 gcactgattg ccaagcgttg ccagcagcag ctgcgccaac gtcagacccg ggtgatcaac
     5161 gccaccggca ccctggtgca caccaatctg gggcgctcgc cgctaagtcg cgagctgtgg
     5221 gacgaggtgc gcgacctcaa cactggctac aacaatctgg aactggatct cgccaccggc
     5281 aagcgcggcg ggcgcaaggg gctgatcgcc cccctgctcc gtgtgctcac ccaggccgag
     5341 gattcgctgg tggtcaacaa caacgccgct tcgctcttct tgctgctgca ggagatagcc
     5401 aaggggcgcg aggtgatcgt ctcgcggggc gaacagatcc agattggtgg cggctttcgc
     5461 attcccgaca ttctggcgct ctccggcgcc aaactggtgg aggtgggcac caccaatatc
     5521 actaccgcca aagattacct cgatgccatc acagatcaga ccgcgctggt gctgatggta
     5581 cacagatcca atttcgccat tcgcggcttt accgaatccc ccgatattgg cgaggtggcc
     5641 cgcgccctgc ccgagcacgt ggtgctggcg gtggatcagg gctcgggctt gaccaccgag
     5701 gagtttgcac cggacgaaac ctcggtgcgt cagtacatca aggcgggggc ggatctggtc
     5761 tgctactccg gcgacaagct gctgggtggc ccgcaatcgg gcatcatcag cggccgcagc
     5821 gacctcatca agcggctgga aaaacacccc atgatgcgca ccttccgccc gagccgcatc
     5881 gtctactccc tgctggaacg cctgctcatc cacaagctca acaagtcccc catcggcgag
     5941 ggcatcgccc agcgcacctt gagcaaccct gccgccatgc aggcccgcgc cgatcagctg
     6001 atggccgccc tgcccggctg ctttgtgccg gtccccgccc agctggtggt gggtggtggc
     6061 accctgccgg acgagttcta ccctgcgcct gcgctcgaat gcaccgaccc gcgtccggcc
     6121 cagcagctgc tcgatgccct gcggaaactg ccggtgccgg tcatcgccac cgtgcgccag
     6181 cagaaggtgc tgctcaatat ggcgaccctg ctgccgaccg agattgcact gcttatcgcc
     6241 caactcaagg agttgctact gcccactccg accactgcga ccgaggagcc ctgatcccac
     6301 agccgccagt tccgctggcg gcattttcga caggctattg tgattcgaac ccgcagagtc
     6361 cttgtggcat aatgccctgt ccaatggccc gctcagggtc atctcctttt gcctgatata
     6421 cgctgtttca tcttgagaaa cagccaaaca aggggtaatc gtgtcttcca ttcgtctgac
     6481 ccaatacagc cacggggctg gctgcggctg caaaatttct cccaaggtgc tcgacaccat
     6541 tctcaagagc cagatcccgg gctttgacga cccgaccctg gtggttggca acagcagcaa
     6601 ggatgacgcg gccgtggtcg atatcggcaa cggtcagggc attgtttcca ccaccgactt
     6661 cttcatgccc atcgtcgatg atccctttac ctttggccgc atcgcggcca ccaacgccat
     6721 cagcgacatc tacgccatgg gcggcaagcc catcgttgcc attgccatcc ttggctggcc
     6781 catcaacacc ctagccccgg aagtggccca gcaggtgata gatggcggcc gccaggtgtg
     6841 ccatgaagcg ggcatatcct tggctggcgg ccacagtatc gatgcccccg agcccatctt
     6901 cggtcttgct gtgaccggta tagtgccgct caatgccatc aagcagaacg acacggccca
     6961 ggcgggtgac atcctctacc tgaccaagcc cctcggtatc ggcatcctca ccacggccca
     7021 gaagaagggc aaattgaagc cagagcatga gcagctggcc cccaacgcca tgtgcaccct
     7081 caacaagatt ggccagcgct ttgccgaact gcccggcgtg cacgccatga cggatgtgac
     7141 cgggtttggc ctggcgggac acctgcttga gatgtgcgaa ggctcagggg tgtgtgccac
     7201 cctcgatttc aaggcgctgc cactgctcga cgaagtagat tactacctgt ccgagggctg
     7261 cgtaccgggc ggtaccctgc gcaacttcga ttcctatggc gccaagctcg gtgccatgga
     7321 tgaacgcacc cgcaacatca tgtgcgatcc gcagaccagc ggcggcttgc tggttgccgt
     7381 cggtaaagaa agtgaagccg agctccttgc tatcgcgaca caagcggggc tgaccctctc
     7441 ccccataggc cagctgaaag cctataccgg aaaccagttt atcgaggtta tccaatgatc
     7501 acacaggaaa cagaccatga ttatggcagt attggatatt acaaatgcta attttgatga
     7561 aaccgttaag accgccaagc ccgttttaat tgacttttgg gcaccgtggt agccgggatg
     7621 cgtacagctc agtcctgagc tgcaggctgc cgaggcggaa ctcggcgaca aggctgtgat
     7681 agcacagtct aacgtggata atgcacgtga attggcagta aaatttaagt ttatgtcaat
     7741 acctaccctc atcgttttaa aagacggaaa agaggtggac aggcacacag gctatatgga
     7801 taagaagagc cttgtaaact ttgtttcaaa gcatatctaa ttcatgagcg gatacatatt
     7861 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7921 acttgcggag acccggtcgt cagcttgtcg tcggttcagg gcggggtcgt taaatagccg
     7981 cttatgtcta ttgctggttt accggtttat tgactaccgg aagcagtgtg accgtgtgct
     8041 tctcaaatgc ctgaggtttc agcaaaaaac ccctcaagac ccgtttagag gcggccgcgg
     8101 ccatccgtca ggatggcctt ctgcttaatt tgatgcctgg cagtttatgg cgggcgtcct
     8161 gcccgccacc ctccgggccg ttgcttcgca acgttcaaat ccgctcccgg cggatttgtc
     8221 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac
     8281 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcatggg gagaccccac
     8341 actaccatcg gcgctacggc gtttcacttc tgagttcggc atggggtcag gtgggaccac
     8401 cgcgctactg ccgccaggca aattctgttt tatcagaccg cttctgcgtt ctgatttaat
     8461 ctgtatcagg ctgctcggat ccatggttaa ttcctaaaaa aaatccttag ctttcgctaa
     8521 ggatctgcag gtcgactcta gatcctggcg gacgggggag gaatcgaacc taacgcctgc
     8581 acagcgtcct gacgggttta gagcccgcga ccgccaccag acggaacccc cgtccgaatt
     8641 caaacgggt
//
