LOCUS       Exported     2504 bp     DNA     circular     SYN     24-JUL-2020
DEFINITION  synthetic linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2001)
  AUTHORS   Keoni Gandall
  TITLE     Direct Submission
  JOURNAL   Exported from Poly.
FEATURES             Location/Qualifiers
     terminator      1..2504
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
 polymerase"
     terminator      517..564
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA polymerase"
     primer_bind     2392..2408
                     /label="M13 for"
                     /note="common sequencing primer, one of multiple similar variants"
     terminator      2409..2440
                     /label="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription terminator"
     CDS             1..0
                     /label="MMSYN1_0776"
                     /note="Required for rescue of stalled ribosomes mediated by trans-translation. Binds to transfer-messenger RNA (tmRNA), required for stable association of tmRNA with ribosomes. tmRNA and SmpB together mimic tRNA shape, replacing the anticodon stem-loop with SmpB. tmRNA is encoded by the ssrA gene; the 2 termini fold to resemble tRNA(Ala) and it encodes a 'tag peptide', a short internal open reading frame. During trans-translation Ala-aminoacylated tmRNA acts like a tRNA, entering the A-site of stalled ribosomes, displacing the stalled mRNA. The ribosome then switches to translate the ORF on the tmRNA; the nascent peptide is terminated with the 'tag peptide' encoded by the tmRNA and targeted for degradation. The ribosome is freed to recommence translation, which seems to be the essential function of trans-translation."
                     /translation="MSEHLIVKNKKAYFNYEIIQTYQAGIVLNGPEIKSIRNHDVSINEAFVLIRKKEIYILNMNVKKYQFANYIKGLEETRTRKLLLHKKEIIKILNKIKQENLTIIPVKLYFKNDYVKLEIALAKGKKLHDKRQTIKKRDTERKELKDYK"
     primer_bind     complement(569..585)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar variants"
     rep_origin      complement(652..1240)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(1411..2271)
                     /label="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     promoter        complement(2272..2376)
                     /label="bla promoter"
                     /note="ampicillin resistance gene promoter"
     CDS             complement(1..0)
                     /label="MMSYN1_0776"
                     /note="Required for rescue of stalled ribosomes mediated by trans-translation. Binds to transfer-messenger RNA (tmRNA), required for stable association of tmRNA with ribosomes. tmRNA and SmpB together mimic tRNA shape, replacing the anticodon stem-loop with SmpB. tmRNA is encoded by the ssrA gene; the 2 termini fold to resemble tRNA(Ala) and it encodes a 'tag peptide', a short internal open reading frame. During trans-translation Ala-aminoacylated tmRNA acts like a tRNA, entering the A-site of stalled ribosomes, displacing the stalled mRNA. The ribosome then switches to translate the ORF on the tmRNA; the nascent peptide is terminated with the 'tag peptide' encoded by the tmRNA and targeted for degradation. The ribosome is freed to recommence translation, which seems to be the essential function of trans-translation."
                     /translation="MSEHLIVKNKKAYFNYEIIQTYQAGIVLNGPEIKSIRNHDVSINEAFVLIRKKEIYILNMNVKKYQFANYIKGLEETRTRKLLLHKKEIIKILNKIKQENLTIIPVKLYFKNDYVKLEIALAKGKKLHDKRQTIKKRDTERKELKDYK"
ORIGIN
        1 ATGTCTGAGC ATTTAATTGT GAAGAATAAA AAGGCGTACT TTAATTACGA GATTATCCAA
       61 ACCTATCAGG CTGGAATTGT TCTTAATGGC CCGGAGATTA AATCAATTCG TAATCATGAC
      121 GTGAGCATTA ACGAAGCCTT CGTACTGATC CGTAAAAAGG AGATTTATAT CCTTAATATG
      181 AATGTGAAGA AGTATCAGTT CGCAAATTAT ATCAAGGGAC TGGAAGAAAC ACGTACTCGC
      241 AAGTTGTTGT TACATAAGAA GGAGATCATT AAGATCCTGA ACAAAATTAA ACAGGAAAAC
      301 TTGACAATCA TCCCAGTAAA ACTTTATTTC AAAAATGATT ATGTCAAGTT GGAGATTGCG
      361 TTAGCCAAGG GGAAGAAATT GCATGACAAA CGCCAGACTA TCAAGAAGCG TGACACTGAA
      421 CGCAAAGAAC TGAAGGACTA CAAGTGAAGA GCTTAGAGAC CGGTACTAGT TCCTGCAGGG
      481 CTCGAGGTGT CAATCGTCGG AGCCGCTGAG CAATAACTAG CATAACCCCT TGGGGCCTCT
      541 AAACGGGTCT TGAGGGGTTT TTTGCATGGT CATAGCTGTT TCCTGAGAGC TTGGCAGGTG
      601 ATGACACACA TTAACAAATT TCGTGAGGAG TCTCCAGAAG AATGCCATTA ATTTCCATAG
      661 GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA AGTCAGAGGT GGCGAAACCC
      721 GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC GCTCTCCTGT
      781 TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA GCGTGGCGCT
      841 TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG GTCGTTCGCT CCAAGCTGGG
      901 CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC TTATCCGGTA ACTATCGTCT
      961 TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA GCAGCCACTG GTAACAGGAT
     1021 TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC CTAACTACGG
     1081 CTACACTAGA AGAACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA CCTTCGGAAA
     1141 AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT GGTAGCGGTG GTTTTTTTGT
     1201 TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA GAAGATCCTT TGATCTTTTC
     1261 TACGGGGTCT GACGCTCAGT GGAACGAAAA CTCACGTTAA GGGATTTTGG TCATGAGATT
     1321 ATCAAAAAGG ATCTTCACCT AGATCCTTTT AAATTAAAAA TGAAGTTTTA AATCAATCTA
     1381 AAGTATATAT GAGTAAACTT GGTCTGACAG TTACCAATGC TTAATCAGTG AGGCACCTAT
     1441 CTCAGCGATC TGTCTATTTC GTTCATCCAT AGTTGCCTGA CTCCCCGTCG TGTAGATAAC
     1501 TACGATACGG GAGGGCTTAC CATCTGGCCC CAGTGCTGCA ATGATACCGC GAGAACCACG
     1561 CTCACCGGCT CCAGATTTAT CAGCAATAAA CCAGCCAGCC GGAAGGGCCG AGCGCAGAAG
     1621 TGGTCCTGCA ACTTTATCCG CCTCCATCCA GTCTATTAAT TGTTGCCGGG AAGCTAGAGT
     1681 AAGTAGTTCG CCAGTTAATA GTTTGCGCAA CGTTGTTGCC ATTGCTACAG GCATCGTGGT
     1741 GTCACGCTCG TCGTTTGGTA TGGCTTCATT CAGCTCCGGT TCCCAACGAT CAAGGCGAGT
     1801 TACATGATCC CCCATGTTGT GCAAAAAAGC GGTTAGCTCC TTCGGTCCTC CGATCGTTGT
     1861 CAGAAGTAAG TTGGCCGCAG TGTTATCACT CATGGTTATG GCAGCACTGC ATAATTCTCT
     1921 TACTGTCATG CCATCCGTAA GATGCTTTTC TGTGACTGGT GAGTACTCAA CCAAGTCATT
     1981 CTGAGAATAG TGTATGCGGC GACCGAGTTG CTCTTGCCCG GCGTCAATAC GGGATAATAC
     2041 CGCGCCACAT AGCAGAACTT TAAAAGTGCT CATCATTGGA AAACGTTCTT CGGGGCGAAA
     2101 ACTCTCAAGG ATCTTACCGC TGTTGAGATC CAGTTCGATG TAACCCACTC GTGCACCCAA
     2161 CTGATCTTCA GCATCTTTTA CTTTCACCAG CGTTTCTGGG TGAGCAAAAA CAGGAAGGCA
     2221 AAATGCCGCA AAAAAGGGAA TAAGGGCGAC ACGGAAATGT TGAATACTCA TACTCTTCCT
     2281 TTTTCAATAT TATTGAAGCA TTTATCAGGG TTATTGTCTC ATGAGCGGAT ACATATTTGA
     2341 ATGTATTTAG AAAAATAAAC AAATAGGGGT TCCGCGCACC TGCACCAGTC AGTAAAACGA
     2401 CGGCCAGTAG TCAAAAGCCT CCGACCGGAG GCTTTTGACT TGGTCAGGTG GAGTGAATTC
     2461 GCGGCCGCTT CTAGAGGCTC TTCGTAATTA CGAAGAGGTC TCTA
//