LOCUS       Exported     3662 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..3662
                     /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..3661
                     /label="lnt"
                     /note="Catalyzes the phospholipid dependent N-acylation of the N-terminal cysteine of apolipoprotein, the last step in lipoprotein maturation (PubMed:2032623, PubMed:21676878, PubMed:28885614, PubMed:28675161). Utilizes a two-step reaction via a ping-pong mechanism (PubMed:21676878, PubMed:28675161). Lnt undergoes covalent modification in the presence of phospholipids, resulting in a thioester acyl-enzyme intermediate. It then transfers the acyl chain to the amine group of the N-terminal diacylglyceryl-modified cysteine of apolipoprotein, leading to the formation of mature triacylated lipoprotein (PubMed:21676878, PubMed:28675161). In vitro, can utilize the phospholipids phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidic acid (PA) or cardiolipin (CL) (PubMed:2032623, PubMed:21676878). PE is the most efficient acyl donor (PubMed:21676878)."
                     /translation="MAFASLIERQRIRLLLALLFGACGTLAFSPYDVWPAAIISLMGLQALTFNRRPLQSAAIGFCWGFGLFGSGINWVYVSIATFGGMPGPVNIFLVVLLAAYLSLYTGLFAGVLSRLWPKTTWLRVAIAAPALWQVTEFLRGWVLTGFPWLQFGYSQIDGPLKGLAPIMGVEAINFLLMMVSGLLALALVKRNWRPLVVAVVLFALPFPLRYIQWFTPQPEKTIQVSMVQGDIPQSLKWDEGQLLNTLKIYYNATAPLMGKSSLIIWPESAITDLEINQQPFLKALDGELRDKGSSLVTGIVDARLNKQNRYDTYNTIITLGKGAPYSYESADRYNKNHLVPFGEFVPLESILRPLAPFFDLPMSSFSRGPYIQPPLSANGIELTAAICYEIILGEQVRDNFRPDTDYLLTISNDAWFGKSIGPWQHFQMARMRALELARPLLRSTNNGITAVIGPQGEIQAMIPQFTREVLTTNVTPTTGLTPYARTGNWPLWVLTALFGFAAVLMSLRQRRK"
     primer_bind     complement(90..106)
                     /note="common sequencing primer, one of multiple similar variants"
                     /label="M13 rev"
     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    3603..3662
                     /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 TGGCATTCGC
     2041 ATCACTAATC GAACGTCAGC GTATCCGTCT ACTACTAGCA CTACTATTCG GAGCATGCGG
     2101 AACCCTAGCA TTCTCACCGT ACGACGTCTG GCCGGCAGCA ATCATCTCAC TAATGGGACT
     2161 ACAGGCACTA ACCTTCAACC GTCGTCCGCT ACAGTCAGCA GCAATCGGAT TCTGCTGGGG
     2221 ATTCGGACTA TTCGGATCAG GAATCAACTG GGTCTACGTC TCAATCGCAA CCTTCGGAGG
     2281 AATGCCGGGA CCGGTCAACA TCTTCCTAGT CGTCCTACTA GCAGCATACC TATCACTATA
     2341 CACCGGACTA TTCGCAGGAG TCCTATCACG TCTATGGCCG AAAACCACCT GGCTACGTGT
     2401 CGCAATCGCA GCACCGGCAC TATGGCAGGT CACCGAATTC CTACGTGGAT GGGTCCTAAC
     2461 CGGATTCCCG TGGCTACAGT TCGGATACTC ACAGATCGAC GGACCGCTAA AAGGACTAGC
     2521 ACCGATCATG GGAGTCGAAG CAATCAACTT CCTACTAATG ATGGTCTCAG GACTACTAGC
     2581 ACTAGCACTA GTCAAACGTA ACTGGCGTCC GCTAGTCGTC GCAGTCGTCC TATTCGCACT
     2641 ACCGTTCCCG CTACGTTACA TCCAGTGGTT CACCCCGCAG CCGGAAAAAA CCATCCAGGT
     2701 CTCAATGGTC CAGGGAGACA TCCCGCAGTC ACTAAAATGG GACGAAGGAC AGCTACTAAA
     2761 CACCCTAAAA ATCTACTACA ACGCAACCGC ACCGCTAATG GGAAAATCAT CACTAATCAT
     2821 CTGGCCGGAA TCAGCAATCA CCGACCTAGA AATCAACCAG CAGCCGTTCC TAAAAGCACT
     2881 AGACGGAGAA CTACGTGACA AAGGATCATC ACTAGTCACC GGAATCGTCG ACGCACGTCT
     2941 AAACAAACAG AACCGTTACG ACACCTACAA CACCATCATC ACCCTAGGAA AAGGAGCACC
     3001 GTACTCATAC GAATCAGCAG ACCGTTACAA CAAAAACCAC CTAGTCCCGT TCGGAGAATT
     3061 CGTCCCGCTA GAATCAATCC TACGTCCGCT AGCACCGTTC TTCGACCTAC CGATGTCATC
     3121 ATTCTCACGT GGACCGTACA TCCAGCCGCC GCTATCAGCA AACGGAATCG AACTAACCGC
     3181 AGCAATCTGC TACGAAATCA TCCTAGGAGA ACAGGTCCGT GACAACTTCC GTCCGGACAC
     3241 CGACTACCTA CTAACCATCT CAAACGACGC ATGGTTCGGA AAATCAATCG GACCGTGGCA
     3301 GCACTTCCAG ATGGCACGTA TGCGTGCACT AGAACTAGCA CGTCCGCTAC TACGTTCAAC
     3361 CAACAACGGA ATCACCGCAG TCATCGGACC GCAGGGAGAA ATCCAGGCAA TGATCCCGCA
     3421 GTTCACCCGT GAAGTCCTAA CCACCAACGT CACCCCGACC ACCGGACTAA CCCCGTACGC
     3481 ACGTACCGGA AACTGGCCGC TATGGGTCCT AACCGCACTA TTCGGATTCG CAGCAGTCCT
     3541 AATGTCACTA CGTCAGCGTC GTAAATGAAG AGCTTCGAGA CCAAATAAGA CGACGACCCT
     3601 CGGCCCACAT ATCCGATAAT GGATAGGGAT AACAGGGTAA TCATACAATG CTATTTCAAA
     3661 AT
//