LOCUS       sequence-316183-        8635 bp    DNA     circular SYN 14-APR-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 8635)
  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 8635)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..8635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     59..78
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(316..333)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      486..572
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(3998..4045)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             complement(5300..6115)
                     /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             complement(5300..6115)
                     /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     6023..6042
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(6116..6207)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6473..6492)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7178..7318
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7244..7263)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7404..7426
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7582..8460)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        join(8487..8635,1..136)
                     /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)"
ORIGIN
        1 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat
       61 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc
      121 tctactgttt ctccataccc gtttgaattc ggacgggggt tccgtctggt ggcggtcgcg
      181 ggctctaaac ccgtcaggac gctgtgcagg cgttaggttc gattcctccc ccgtccgcca
      241 ggatctagag tcgacctgca gatccttagc gaaagctaag gatttttttt aggaattaac
      301 catggatccg agcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
      361 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
      421 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca
      481 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
      541 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
      601 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
      661 agcagaaggc catcctgacg gatggccgcg gccgcctcta aacgggtctt gaggggtttt
      721 ttgctgaaac ctcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa
      781 ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga accgacgaca
      841 agctgacgac cgggtctccg caagtggcac ttttcgggga aatgtgcgcg gaacccctat
      901 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaga tatgctttga
      961 aacaaagttt acaaggctct tcttatccat atagcctgtg tgcctgtcca cctcttttcc
     1021 gtcttttaaa acgatgaggg taggtattga cataaactta aattttactg ccaattcacg
     1081 tgcattatcc acgttagact gtgctatcac agccttgtcg ccgagttccg cctcggcagc
     1141 ctgcagctca ggactgagct gtacgcatcc cggctaccac ggtgcccaaa agtcaattaa
     1201 aacgggcttg gcggtcttaa cggtttcatc aaaattagca tttgtaatat ccaatactgc
     1261 cataatcatg gtctgtttcc tgtgtgatca ttggataacc tcgataaact ggtttccggt
     1321 ataggctttc agctggccta tgggggagag ggtcagcccc gcttgtgtcg cgatagcaag
     1381 gagctcggct tcactttctt taccgacggc aaccagcaag ccgccgctgg tctgcggatc
     1441 gcacatgatg ttgcgggtgc gttcatccat ggcaccgagc ttggcgccat aggaatcgaa
     1501 gttgcgcagg gtaccgcccg gtacgcagcc ctcggacagg tagtaatcta cttcgtcgag
     1561 cagtggcagc gccttgaaat cgagggtggc acacacccct gagccttcgc acatctcaag
     1621 caggtgtccc gccaggccaa acccggtcac atccgtcatg gcgtgcacgc cgggcagttc
     1681 ggcaaagcgc tggccaatct tgttgagggt gcacatggcg ttgggggcca gctgctcatg
     1741 ctctggcttc aatttgccct tcttctgggc cgtggtgagg atgccgatac cgaggggctt
     1801 ggtcaggtag aggatgtcac ccgcctgggc cgtgtcgttc tgcttgatgg cattgagcgg
     1861 cactataccg gtcacagcaa gaccgaagat gggctcgggg gcatcgatac tgtggccgcc
     1921 agccaaggat atgcccgctt catggcacac ctggcggccg ccatctatca cctgctgggc
     1981 cacttccggg gctagggtgt tgatgggcca gccaaggatg gcaatggcaa cgatgggctt
     2041 gccgcccatg gcgtagatgt cgctgatggc gttggtggcc gcgatgcggc caaaggtaaa
     2101 gggatcatcg acgatgggca tgaagaagtc ggtggtggaa acaatgccct gaccgttgcc
     2161 gatatcgacc acggccgcgt catccttgct gctgttgcca accaccaggg tcgggtcgtc
     2221 aaagcccggg atctggctct tgagaatggt gtcgagcacc ttgggagaaa ttttgcagcc
     2281 gcagccagcc ccgtggctgt attgggtcag acgaatggaa gacacgatta ccccttgttt
     2341 ggctgtttct caagatgaaa cagcgtatat caggcaaaag gagatgaccc tgagcgggcc
     2401 attggacagg gcattatgcc acaaggactc tgcgggttcg aatcacaata gcctgtcgaa
     2461 aatgccgcca gcggaactgg cggctgtggg atcagggctc ctcggtcgca gtggtcggag
     2521 tgggcagtag caactccttg agttgggcga taagcagtgc aatctcggtc ggcagcaggg
     2581 tcgccatatt gagcagcacc ttctgctggc gcacggtggc gatgaccggc accggcagtt
     2641 tccgcagggc atcgagcagc tgctgggccg gacgcgggtc ggtgcattcg agcgcaggcg
     2701 cagggtagaa ctcgtccggc agggtgccac cacccaccac cagctgggcg gggaccggca
     2761 caaagcagcc gggcagggcg gccatcagct gatcggcgcg ggcctgcatg gcggcagggt
     2821 tgctcaaggt gcgctgggcg atgccctcgc cgatggggga cttgttgagc ttgtggatga
     2881 gcaggcgttc cagcagggag tagacgatgc ggctcgggcg gaaggtgcgc atcatggggt
     2941 gtttttccag ccgcttgatg aggtcgctgc ggccgctgat gatgcccgat tgcgggccac
     3001 ccagcagctt gtcgccggag tagcagacca gatccgcccc cgccttgatg tactgacgca
     3061 ccgaggtttc gtccggtgca aactcctcgg tggtcaagcc cgagccctga tccaccgcca
     3121 gcaccacgtg ctcgggcagg gcgcgggcca cctcgccaat atcgggggat tcggtaaagc
     3181 cgcgaatggc gaaattggat ctgtgtacca tcagcaccag cgcggtctga tctgtgatgg
     3241 catcgaggta atctttggcg gtagtgatat tggtggtgcc cacctccacc agtttggcgc
     3301 cggagagcgc cagaatgtcg ggaatgcgaa agccgccacc aatctggatc tgttcgcccc
     3361 gcgagacgat cacctcgcgc cccttggcta tctcctgcag cagcaagaag agcgaagcgg
     3421 cgttgttgtt gaccaccagc gaatcctcgg cctgggtgag cacacggagc aggggggcga
     3481 tcagcccctt gcgcccgccg cgcttgccgg tggcgagatc cagttccaga ttgttgtagc
     3541 cagtgttgag gtcgcgcacc tcgtcccaca gctcgcgact tagcggcgag cgccccagat
     3601 tggtgtgcac cagggtgccg gtggcgttga tcacccgggt ctgacgttgg cgcagctgct
     3661 gctggcaacg cttggcaatc agtgcctcga tttgctcggg ggcaacccca tgctggcgaa
     3721 atgcctcgct ctggcgcaat tcgctcagga catcgcgcac cgccgaggtc acaaaagggc
     3781 ggctcagcgc ctcgataaaa ccggtgagaa agggttgctg cagcagctgt tccacttgcg
     3841 gtagacggcg cgcttgttgc tggctgggct gtggcagtga atctggcagt gaatctggca
     3901 gtgaatcgtc ggcagtggga catgattcgg gctgactgtg agagtgggcg atggctggcg
     3961 cgtgagacga gttcgtcacg gtttagttcc tcaccttgtc gtattatact atgccgatat
     4021 actatgccga tgattaattg tcaacacgtg attatcccag caaacggtga atacgttgca
     4081 ggccggtcac cagacgatcc acttcttcat gggtgttata catggccagc gacgcccgac
     4141 acatcgcagg gacgttgtaa taggccatca atggcattgc ggcgtgatgt ccggtacgca
     4201 cagcaatgcc gtaattatcg agaaaactgc caacatcata ggcgtggtgt ttaccgagat
     4261 taaaagcaat aacgccaagc ctgttttgtg ggccatagag agtgagatcc ggtacagatt
     4321 ccagctgtga tagcgcataa tgcatcagat tctgttcata ctcggctatg ttattaagcc
     4381 ccagcgccga aacatactcc agcgccgcgc caagaccaat gatgcccccg gtattgggtg
     4441 taccggcttc aaaccgccat ggtgctttgg tccaggtagt gccttcactc aggctgacgg
     4501 tggcgatcat agaaccgccc ccttcccacg gcggcatctc ctgcaacaag gcttctttca
     4561 cataaagaat gccaattccg gtggggccat acagtttatg cccggagaac acgtaaaagt
     4621 cgcaatccag cgcctgaaca tccaccggat gatgcatcac cgcctgagcg ccatccacca
     4681 gcacttttgc gccatgctgg tgcgcaagcg tgatcatttc cgccagtgga ttttctgtgc
     4741 caagcacgtt ggagacatga gtaattgcca gcaggcgagt tttctcatca aacagcgtag
     4801 gcagcgtctc cagttgcaac gtaccatcgg gattgagcgg gatcacacgc agctctgcgc
     4861 caacgcgtgc gcaaagcatc tgccagggaa caatgttagc gtggtgctcc atctgactga
     4921 tgatgatgtt atcgcccgcc cgcacgttgc tgttgcccca gctattggcg accagattga
     4981 tcccttccgt cgtgccgcgg acgaacacca gctcttccgc cgaacgggca ttaataaaca
     5041 gcgatgcccg cttgcgcacg ttctccattt tctcggtcgc ctgggcgctt aaggtatgaa
     5101 taccacgatg caccgccgcg tagccatgac gataaaactc ggcctcggcg tcaatcacct
     5161 ggctcggttt ctgcgcactg gcggcgctgt cgagataagc cagcggcaaa ccgtttacct
     5221 cacgcgaaag caccggaaag tcggcccgca ctttgtcgac ggaaaaaatc atcttgcacc
     5281 tcctggcagt tattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
     5341 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
     5401 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
     5461 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
     5521 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
     5581 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
     5641 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcggtcgctg ttaaaaggac
     5701 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
     5761 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
     5821 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
     5881 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     5941 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     6001 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     6061 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcatactct
     6121 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6181 ttgaatgtat ttagaaaaat aaacaaatag gcatgcagcg ctcttccgct tcctcgctca
     6241 ctgactcgct acgctcggtc gttcgactgc ggcgagcggt gtcagctcac tcaaaagcgg
     6301 taatacggtt atccacagaa tcaggggata aagccggaaa gaacatgtga gcaaaaagca
     6361 aagcaccgga agaagccaac gccgcaggcg tttttccata ggctccgccc ccctgacgag
     6421 catcacaaaa atcgacgctc aagccagagg tggcgaaacc cgacaggact ataaagatac
     6481 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6541 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6601 tggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6661 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6721 cacgacttat cgccactggc agcagccatt ggtaactgat ttagaggact ttgtcttgaa
     6781 gttatgcacc tgttaaggct aaactgaaag aacagatttt ggtgagtgcg gtcctccaac
     6841 ccacttacct tggttcaaag agttggtagc tcagcgaacc ttgagaaaac caccgttggt
     6901 agcggtggtt tttctttatt tatgagatga tgaatcaatc ggtctatcaa gtcaacgaac
     6961 agctattccg ggccggccgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7021 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7081 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7141 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     7201 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     7261 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     7321 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     7381 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     7441 accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc atgcataatg
     7501 tgcctgtcaa atggacgaag cagggattct gcaaacccta tgctactccg tcaagccgtc
     7561 aattgtctga ttcgttacca attatgacaa cttgacggct acatcattca ctttttcttc
     7621 acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata cccgcgagaa
     7681 atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca tccgggtggt
     7741 gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta agacgctaat
     7801 ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa catgctgtgc
     7861 gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact gacaagcctc
     7921 gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca tgcgccgcag
     7981 taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt ccccttgccc
     8041 ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt catccgggcg
     8101 aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc agtaggcgcg
     8161 cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac gaccgtagtg
     8221 atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa attctcgtcc
     8281 ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac ctttcattcc
     8341 cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg ttaaacccgc
     8401 caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg cttcagccat
     8461 acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca tcagacattg
     8521 ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc cgcttattaa
     8581 aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca aaagt
//
