LOCUS       Exported     2636 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..2636
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
 polymerase"
     terminator      38..85
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA polymerase"
     primer_bind     1933..1949
                     /label="M13 for"
                     /note="common sequencing primer, one of multiple similar variants"
     terminator      1950..1981
                     /note="bidirectional E. coli tonB-P14 transcription terminator"
                     /label="tonB terminator"
     CDS             2030..2542
                     /label="secM"
                     /note="Regulates secA expression by translational coupling of the secM secA operon. Ribosomes translating the C-terminal region of secM can disrupt an RNA repressor helix that normally blocks secA translation initiation, derepressing the expression of secA. Translational pausing of secM at Pro-166 under secretion-limiting conditions increases the duration of the disruption and thus increases secA expression. This is controlled by interaction of the secM signal peptide with secA and the translocon, possibly by secA pulling the paused secM out of the ribosome. The arrest sequence (150-FXXXXWIXXXXGIRAGP-166) is sufficient to cause arrest of unrelated proteins. Elongation arrest can be alleviated by mutations in the 23S rRNA or in ribosomal protein L22."
                     /translation="MSGILTRWRQFGKRYFWPHLLLGMVAASLGLPALSNAAEPNAPAKATTRNHEPSAKVNFGQLALLEANTRRPNSNYSVDYWHQHAIRTVIRHLSFAMAPQTLPVAEESLPLQAQHLALLDTLSALLTQEGTPSEKGYRIDYAHFTPQAKFSTPVWISQAQGIRAGPQRLT"
     primer_bind     complement(90..106)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar variants"
     rep_origin      complement(173..761)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(952..1812)
                     /label="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     promoter        complement(1813..1917)
                     /label="bla promoter"
                     /note="ampicillin resistance gene promoter"
     misc_feature    2577..2636
                     /label="FreeGenes Barcode"
                     /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences."
ORIGIN
        1 CGCTGAGGTG TCAATCGTCG GAGCCGCTGA GCAATAACTA GCATAACCCC TTGGGGCCTC
       61 TAAACGGGTC TTGAGGGGTT TTTTGCATGG TCATAGCTGT TTCCTGAGAG CTTGGCAGGT
      121 GATGACACAC ATTAACAAAT TTCGTGAGGA GTCTCCAGAA GAATGCCATT AATTTCCATA
      181 GGCTCCGCCC CCCTGACGAG CATCACAAAA ATCGACGCTC AAGTCAGAGG TGGCGAAACC
      241 CGACAGGACT ATAAAGATAC CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG CGCTCTCCTG
      301 TTCCGACCCT GCCGCTTACC GGATACCTGT CCGCCTTTCT CCCTTCGGGA AGCGTGGCGC
      361 TTTCTCATAG CTCACGCTGT AGGTATCTCA GTTCGGTGTA GGTCGTTCGC TCCAAGCTGG
      421 GCTGTGTGCA CGAACCCCCC GTTCAGCCCG ACCGCTGCGC CTTATCCGGT AACTATCGTC
      481 TTGAGTCCAA CCCGGTAAGA CACGACTTAT CGCCACTGGC AGCAGCCACT GGTAACAGGA
      541 TTAGCAGAGC GAGGTATGTA GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG CCTAACTACG
      601 GCTACACTAG AAGAACAGTA TTTGGTATCT GCGCTCTGCT GAAGCCAGTT ACCTTCGGAA
      661 AAAGAGTTGG TAGCTCTTGA TCCGGCAAAC AAACCACCGC TGGTAGCGGT GGTTTTTTTG
      721 TTTGCAAGCA GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGGCCTA CTATTAGCAA
      781 CAACGATCCT TTGATCTTTT CTACGGGGTC TGACGCTCAG TGGAACGAAA ACTCACGTTA
      841 AGGGATTTTG GTCATGAGAT TATCAAAAAG GATCTTCACC TAGATCCTTT TAAATTAAAA
      901 ATGAAGTTTT AAATCAATCT AAAGTATATA TGAGTAAACT TGGTCTGACA GTTACCAATG
      961 CTTAATCAGT GAGGCACCTA TCTCAGCGAT CTGTCTATTT CGTTCATCCA TAGTTGCCTG
     1021 ACTCCCCGTC GTGTAGATAA CTACGATACG GGAGGGCTTA CCATCTGGCC CCAGTGCTGC
     1081 AATGATACCG CGAGAACCAC GCTCACCGGC TCCAGATTTA TCAGCAATAA ACCAGCCAGC
     1141 CGGAAGGGCC GAGCGCAGAA GTGGTCCTGC AACTTTATCC GCCTCCATCC AGTCTATTAA
     1201 TTGTTGCCGG GAAGCTAGAG TAAGTAGTTC GCCAGTTAAT AGTTTGCGCA ACGTTGTTGC
     1261 CATTGCTACA GGCATCGTGG TGTCACGCTC GTCGTTTGGT ATGGCTTCAT TCAGCTCCGG
     1321 TTCCCAACGA TCAAGGCGAG TTACATGATC CCCCATGTTG TGCAAAAAAG CGGTTAGCTC
     1381 CTTCGGTCCT CCGATCGTTG TCAGAAGTAA GTTGGCCGCA GTGTTATCAC TCATGGTTAT
     1441 GGCAGCACTG CATAATTCTC TTACTGTCAT GCCATCCGTA AGATGCTTTT CTGTGACTGG
     1501 TGAGTACTCA ACCAAGTCAT TCTGAGAATA GTGTATGCGG CGACCGAGTT GCTCTTGCCC
     1561 GGCGTCAATA CGGGATAATA CCGCGCCACA TAGCAGAACT TTAAAAGTGC TCATCATTGG
     1621 AAAACGTTCT TCGGGGCGAA AACTCTCAAG GATCTTACCG CTGTTGAGAT CCAGTTCGAT
     1681 GTAACCCACT CGTGCACCCA ACTGATCTTC AGCATCTTTT ACTTTCACCA GCGTTTCTGG
     1741 GTGAGCAAAA ACAGGAAGGC AAAATGCCGC AAAAAAGGGA ATAAGGGCGA CACGGAAATG
     1801 TTGAATACTC ATACTCTTCC TTTTTCAATA TTATTGAAGC ATTTATCAGG GTTATTGTCT
     1861 CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGGG TTCCGCGCAC
     1921 CTGCACCAGT CAGTAAAACG ACGGCCAGTA GTCAAAAGCC TCCGACCGGA GGCTTTTGAC
     1981 TTGGTTCAGG TGGAGTGGGA GAAACCGGAG CCATACAGTA CGGTCTCTAA TGTCAGGAAT
     2041 CCTAACCCGT TGGCGTCAGT TCGGAAAACG TTACTTCTGG CCGCACCTAC TACTAGGAAT
     2101 GGTCGCAGCA TCACTAGGAC TACCGGCACT ATCAAACGCA GCAGAACCGA ACGCACCGGC
     2161 AAAAGCAACC ACCCGTAACC ACGAACCGTC AGCAAAAGTC AACTTCGGAC AGCTAGCACT
     2221 ACTAGAAGCA AACACCCGTC GTCCGAACTC AAACTACTCA GTCGACTACT GGCACCAGCA
     2281 CGCAATCCGT ACCGTCATCC GTCACCTATC ATTCGCAATG GCACCGCAGA CCCTACCGGT
     2341 CGCAGAAGAA TCACTACCGC TACAGGCACA GCACCTAGCA CTACTAGACA CCCTATCAGC
     2401 ACTACTAACC CAGGAAGGAA CCCCGTCAGA AAAAGGATAC CGTATCGACT ACGCACACTT
     2461 CACCCCGCAG GCAAAATTCT CAACCCCGGT CTGGATCTCA CAGGCACAGG GAATCCGTGC
     2521 AGGACCGCAG CGTCTAACCT GAAGAGCTTC GAGACCAAGA ATTACTGACC CCTCGGAACG
     2581 ACCGTACGGT GGTTAGTTAG GGATAACAGG GTAATTTCAC TAAATTACTG ACCAGA
//