LOCUS       Exported     3476 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..3476
                     /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
                     /label="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription terminator"
     CDS             100..3475
                     /label="rseP"
                     /note="A site-2 regulated intramembrane protease (S2P) that cleaves the peptide bond between 'Ala-108' and 'Cys-109' in the transmembrane region of RseA. Part of a regulated intramembrane proteolysis (RIP) cascade. Acts on DegS-cleaved RseA to release the cytoplasmic domain of RseA, residue 'Val-148' of RseA may be required for this. This provides the cell with sigma-E (RpoE) activity through the proteolysis of RseA. Can also cleave sequences in transmembrane regions of other proteins (such as LacY) as well as liberated signal peptides of beta-lactamase, OmpF, LivK, SecM, PhoA, LivJ, OmpC, Lpp and TorA, probably within the membrane."
                     /translation="MLSFLWDLASFIVALGVLITVHEFGHFWVARRCGVRVERFSIGFGKALWRRTDKLGTEYVIALIPLGGYVKMLDERAEPVVPELRHHAFNNKSVGQRAAIIAAGPVANFIFAIFAYWLVFIIGVPGVRPVVGEIAANSIAAEAQIAPGTELKAVDGIETPDWDAVRLQLVDKIGDESTTITVAPFGSDQRRDVKLDLRHWAFEPDKEDPVSSLGIRPRGPQIEPVLENVQPNSAASKAGLQAGDRIVKVDGQPLTQWVTFVMLVRDNPGKSLALEIERQGSPLSLTLIPESKPGNGKAIGFVGIEPKVIPLPDEYKVVRQYGPFNAIVEATDKTWQLMKLTVSMLGKLITGDVKLNNLSGPISIAKGAGMTAELGVVYYLPFLALISVNLGIINLFPLPVLDGGHLLFLAIEKIKGGPVSERVQDFCYRIGSILLVLLMGLALFNDFSRL"
     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    3417..3476
                     /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 GAAACACGTG GCAAACATTC CGGTCTCTAA TGCTATCATT
     2041 CCTATGGGAC CTAGCATCAT TCATCGTCGC ACTAGGAGTC CTAATCACCG TCCACGAATT
     2101 CGGACACTTC TGGGTCGCAC GTCGTTGCGG AGTCCGTGTC GAACGTTTCT CAATCGGATT
     2161 CGGAAAAGCA CTATGGCGTC GTACCGACAA ACTAGGAACC GAATACGTCA TCGCACTAAT
     2221 CCCGCTAGGA GGATACGTCA AAATGCTAGA CGAACGTGCA GAACCGGTCG TCCCGGAACT
     2281 ACGTCACCAC GCATTCAACA ACAAATCAGT CGGACAGCGT GCAGCAATCA TCGCAGCAGG
     2341 ACCGGTCGCA AACTTCATCT TCGCAATCTT CGCATACTGG CTAGTCTTCA TCATCGGAGT
     2401 CCCGGGAGTC CGTCCGGTCG TCGGAGAAAT CGCAGCAAAC TCAATCGCAG CAGAAGCACA
     2461 GATCGCACCG GGAACCGAAC TAAAAGCAGT CGACGGAATC GAAACCCCGG ACTGGGACGC
     2521 AGTCCGTCTA CAGCTAGTCG ACAAAATCGG AGACGAATCA ACCACCATCA CCGTCGCACC
     2581 GTTCGGATCA GACCAGCGTC GTGACGTCAA ACTAGACCTA CGTCACTGGG CATTCGAACC
     2641 GGACAAAGAA GACCCGGTCT CATCACTAGG AATCCGTCCG CGTGGACCGC AGATCGAACC
     2701 GGTCCTAGAA AACGTCCAGC CGAACTCAGC AGCATCAAAA GCAGGACTAC AGGCAGGAGA
     2761 CCGTATCGTC AAAGTCGACG GACAGCCGCT AACCCAGTGG GTCACCTTCG TCATGCTAGT
     2821 CCGTGACAAC CCGGGAAAAT CACTAGCACT AGAAATCGAA CGTCAGGGAT CACCGCTATC
     2881 ACTAACCCTA ATCCCGGAAT CAAAACCGGG AAACGGAAAA GCAATCGGAT TCGTCGGAAT
     2941 CGAACCGAAA GTCATCCCGC TACCGGACGA ATACAAAGTC GTCCGTCAGT ACGGACCGTT
     3001 CAACGCAATC GTCGAAGCAA CCGACAAAAC CTGGCAGCTA ATGAAACTAA CCGTCTCAAT
     3061 GCTAGGAAAA CTAATCACCG GAGACGTCAA ACTAAACAAC CTATCAGGAC CGATCTCAAT
     3121 CGCAAAAGGA GCAGGAATGA CCGCAGAACT AGGAGTCGTC TACTACCTAC CGTTCCTAGC
     3181 ACTAATCTCA GTCAACCTAG GAATCATCAA CCTATTCCCG CTACCGGTCC TAGACGGAGG
     3241 ACACCTACTA TTCCTAGCAA TCGAAAAAAT CAAAGGAGGA CCGGTCTCAG AACGTGTCCA
     3301 GGACTTCTGC TACCGTATCG GATCAATCCT ACTAGTCCTA CTAATGGGAC TAGCACTATT
     3361 CAACGACTTC TCACGTCTAT GAAGAGCTTC GAGACCCCGG GAGGAAGATA TAGCACGGGT
     3421 TAAGCAAAAA GCAGCTCTAG GGATAACAGG GTAATTTCCT AGGACGCACC TCATAC
//