LOCUS       sequence-471093-        7383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses machinery necessary for selenocysteine incorporation at 
            UAG codons in BL21-derived cells, spectinomycin resistant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7383)
  TITLE     pSecUAG_UTu1D
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7383)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSecUAG_UTu1D
FEATURES             Location/Qualifiers
     source          1..7383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(355..374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1074..1214
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1140..1159)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1300..1322
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1478..2356)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2383..2667
                     /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     2590..2609
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     complement(2847..2864)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3017..3103
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(6529..6576)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             complement(6589..7380)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(join(7381..7383,1..89))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
       61 atttgaatgt atttagaaaa ataaacaaat aggcatgcag cgctcttccg cttcctcgct
      121 cactgactcg ctacgctcgg tcgttcgact gcggcgagcg gtgtcagctc actcaaaagc
      181 ggtaatacgg ttatccacag aatcagggga taaagccgga aagaacatgt gagcaaaaag
      241 caaagcaccg gaagaagcca acgccgcagg cgtttttcca taggctccgc ccccctgacg
      301 agcatcacaa aaatcgacgc tcaagccaga ggtggcgaaa cccgacagga ctataaagat
      361 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      421 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      481 gttggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      541 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      601 gacacgactt atcgccactg gcagcagcca ttggtaactg atttagagga ctttgtcttg
      661 aagttatgca cctgttaagg ctaaactgaa agaacagatt ttggtgagtg cggtcctcca
      721 acccacttac cttggttcaa agagttggta gctcagcgaa ccttgagaaa accaccgttg
      781 gtagcggtgg tttttcttta tttatgagat gatgaatcaa tcggtctatc aagtcaacga
      841 acagctattc cgggccggcc gatttcggcc ggccggagcc tatggaaaaa cgccagcaac
      901 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      961 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     1021 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     1081 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     1141 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
     1201 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     1261 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1321 ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga
     1381 agcggcatgc ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct
     1441 actccgtcaa gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat
     1501 cattcacttt ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt
     1561 taaatacccg cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga
     1621 taggcatccg ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc
     1681 agcttaagac gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca
     1741 agcaaacatg ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga
     1801 tgtactgaca agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg
     1861 cttccatgcg ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc
     1921 gcccttcccc ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt
     1981 gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt
     2041 catgccagta ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg
     2101 gatgacgacc gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc
     2161 aaacaaattc tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa
     2221 tataaccttt cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa
     2281 tcggcgttaa acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat
     2341 tttgcgcttc agccatactt ttcatactcc cgccattcag agaagaaacc aattgtccat
     2401 attgcatcag acattgccgt cactgcgtct tttactggct cttctcgcta accaaaccgg
     2461 taaccccgct tattaaaagc attctgtaac aaagcgggac caaagccatg acaaaaacgc
     2521 gtaacaaaag tgtctataat cacggcagaa aagtccacat tgattatttg cacggcgtca
     2581 cactttgcta tgccatagca tttttatcca taagattagc ggatcctacc tgacgctttt
     2641 tatcgcaact ctctactgtt tctccatacc cgtttgaatt cggaggggaa cttctgtctg
     2701 gtggcagacg ggaactctaa attccttgaa atgcctcgcc gcattgggtt cgattccctt
     2761 cccctccgcc aggatctaga gtcgacctgc agatccttag cgaaagctaa ggattttttt
     2821 taggaattaa ccatggatcc gagcagcctg atacagatta aatcagaacg cagaagcggt
     2881 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     2941 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     3001 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     3061 tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt
     3121 gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag
     3181 gcatcaaatt aagcagaagg ccatcctgac ggatggccgc ggccgcctct aaacgggtct
     3241 tgaggggttt tttgctgaaa cctcaggcat ttgagaagca cacggtcaca ctgcttccgg
     3301 tagtcaataa accggtaaac cagcaataga cataagcggc tatttaacga ccccgccctg
     3361 aaccgacgac aagctgacga ccgggtctcc gcaagtggca cttttcgggg aaatgtgcgc
     3421 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattag
     3481 atatgctttg aaacaaagtt tacaaggctc ttcttatcca tatagcctgt gtgcctgtcc
     3541 acctcttttc cgtcttttaa aacgatgagg gtaggtattg acataaactt aaattttact
     3601 gccaattcac gtgcattatc cacgttagac tgtgctatca cagccttgtc gccgagttcc
     3661 gcctcggcag cctgcagctc aggactgagc tgtacgcatc ccggctacca cggtgcccaa
     3721 aagtcaatta aaacgggctt ggcggtctta acggtttcat caaaattagc atttgtaata
     3781 tccaatactg ccataatcat ggtctgtttc ctgtgtgatc attggataac ctcgataaac
     3841 tggtttccgg tataggcttt cagctggcct atgggggaga gggtcagccc cgcttgtgtc
     3901 gcgatagcaa ggagctcggc ttcactttct ttaccgacgg caaccagcaa gccgccgctg
     3961 gtctgcggat cgcacatgat gttgcgggtg cgttcatcca tggcaccgag cttggcgcca
     4021 taggaatcga agttgcgcag ggtaccgccc ggtacgcagc cctcggacag gtagtaatct
     4081 acttcgtcga gcagtggcag cgccttgaaa tcgagggtgg cacacacccc tgagccttcg
     4141 cacatctcaa gcaggtgtcc cgccaggcca aacccggtca catccgtcat ggcgtgcacg
     4201 ccgggcagtt cggcaaagcg ctggccaatc ttgttgaggg tgcacatggc gttgggggcc
     4261 agctgctcat gctctggctt caatttgccc ttcttctggg ccgtggtgag gatgccgata
     4321 ccgaggggct tggtcaggta gaggatgtca cccgcctggg ccgtgtcgtt ctgcttgatg
     4381 gcattgagcg gcactatacc ggtcacagca agaccgaaga tgggctcggg ggcatcgata
     4441 ctgtggccgc cagccaagga tatgcccgct tcatggcaca cctggcggcc gccatctatc
     4501 acctgctggg ccacttccgg ggctagggtg ttgatgggcc agccaaggat ggcaatggca
     4561 acgatgggct tgccgcccat ggcgtagatg tcgctgatgg cgttggtggc cgcgatgcgg
     4621 ccaaaggtaa agggatcatc gacgatgggc atgaagaagt cggtggtgga aacaatgccc
     4681 tgaccgttgc cgatatcgac cacggccgcg tcatccttgc tgctgttgcc aaccaccagg
     4741 gtcgggtcgt caaagcccgg gatctggctc ttgagaatgg tgtcgagcac cttgggagaa
     4801 attttgcagc cgcagccagc cccgtggctg tattgggtca gacgaatgga agacacgatt
     4861 accccttgtt tggctgtttc tcaagatgaa acagcgtata tcaggcaaaa ggagatgacc
     4921 ctgagcgggc cattggacag ggcattatgc cacaaggact ctgcgggttc gaatcacaat
     4981 agcctgtcga aaatgccgcc agcggaactg gcggctgtgg gatcagggct cctcggtcgc
     5041 agtggtcgga gtgggcagta gcaactcctt gagttgggcg ataagcagtg caatctcggt
     5101 cggcagcagg gtcgccatat tgagcagcac cttctgctgg cgcacggtgg cgatgaccgg
     5161 caccggcagt ttccgcaggg catcgagcag ctgctgggcc ggacgcgggt cggtgcattc
     5221 gagcgcaggc gcagggtaga actcgtccgg cagggtgcca ccacccacca ccagctgggc
     5281 ggggaccggc acaaagcagc cgggcagggc ggccatcagc tgatcggcgc gggcctgcat
     5341 ggcggcaggg ttgctcaagg tgcgctgggc gatgccctcg ccgatggggg acttgttgag
     5401 cttgtggatg agcaggcgtt ccagcaggga gtagacgatg cggctcgggc ggaaggtgcg
     5461 catcatgggg tgtttttcca gccgcttgat gaggtcgctg cggccgctga tgatgcccga
     5521 ttgcgggcca cccagcagct tgtcgccgga gtagcagacc agatccgccc ccgccttgat
     5581 gtactgacgc accgaggttt cgtccggtgc aaactcctcg gtggtcaagc ccgagccctg
     5641 atccaccgcc agcaccacgt gctcgggcag ggcgcgggcc acctcgccaa tatcggggga
     5701 ttcggtaaag ccgcgaatgg cgaaattgga tctgtgtacc atcagcacca gcgcggtctg
     5761 atctgtgatg gcatcgaggt aatctttggc ggtagtgata ttggtggtgc ccacctccac
     5821 cagtttggcg ccggagagcg ccagaatgtc gggaatgcga aagccgccac caatctggat
     5881 ctgttcgccc cgcgagacga tcacctcgcg ccccttggct atctcctgca gcagcaagaa
     5941 gagcgaagcg gcgttgttgt tgaccaccag cgaatcctcg gcctgggtga ggcaacggag
     6001 caggggggcg atcagcccct tgcgcccgcc gcgcttgccg gtggcgagat ccagttccag
     6061 attgttgtag ccagtgttga ggtcgcgcac ctcgtcccac agctcgcgac ttagcggcga
     6121 gcgccccaga ttggtgtgca ccagggtgcc ggtggcgttg atcacccggg tctgacgttg
     6181 gcgcagctgc tgctggcaac gcttggcaat cagtgcctcg atttgctcgg gggcaacccc
     6241 atgctggcga aatgcctcgc tctggcgcaa ttcgctcagg acatcgcgca ccgcctgggt
     6301 caccagcggg cggctcagcg cctcgataaa accggtgaga aagggttgct gcagcagctg
     6361 ttccacttgc ggtagacggc gcgcttgttg ctggctgggc tgtggcagtg aatctggcag
     6421 tgaatctggc agtgaatcgt cggcagtggg acatgattcg ggctgactgt gagagtgggc
     6481 gatggctggc gcgtgagacg agttcggcat ggtttagttc ctcaccttgt cgtattatac
     6541 tatgccgata tactatgccg atgattaatt gtcaacacgt gttaattctt atttgccgac
     6601 taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat ctgcgcgcga
     6661 ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg
     6721 atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt
     6781 gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc
     6841 agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc
     6901 ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg
     6961 ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat
     7021 acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata
     7081 acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc
     7141 gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt
     7201 ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc
     7261 gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc
     7321 gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat
     7381 act
//
