LOCUS Exported 3224 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..3224 /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..3223 /label="dnaN" /note="Confers DNA tethering and processivity to DNA polymerases and other proteins. Acts as a clamp, forming a ring around DNA (a reaction catalyzed by the clamp-loading complex) which diffuses in an ATP-independent manner freely and bidirectionally along dsDNA (PubMed:2040637). DNA bound in the ring is bent 22 degrees, in solution primed DNA is bound more tightly than dsDNA, suggesting the clamp binds both ss- and dsDNA (PubMed:18191219). In a complex of DNA with this protein, alpha, epsilon and tau subunits however the DNA is only slightly bent (PubMed:26499492). Coordinates protein traffic at the replication fork, where it interacts with multiple DNA polymerases, repair factors and other proteins (PubMed:15466025, PubMed:16168375, PubMed:22716942, PubMed:14592985, PubMed:14729336, PubMed:26499492, PubMed:15952889). Initially characterized for its ability to contact the alpha subunit (dnaE) of DNA polymerase III (Pol III), tethering it to the DNA and conferring very high processivity (PubMed:2040637). Pol III is a complex, multichain enzyme responsible for most of the replicative synthesis in bacteria; it also exhibits 3'-5' exonuclease proofreading activity. The beta chain is required for initiation of replication as well as for processivity of DNA replication (PubMed:3519609, PubMed:2040637). A single clamp can bind both Pol III and IV, allowing the repair Pol IV to access DNA when it is damaged and needs to be fixed, a process the replicative polymerase cannot perform; when DNA is repaired Pol III takes over again (PubMed:16168375). Serves as a processivity factor for DNA polymerases II (PubMed:1999435, PubMed:1534562), IV (PubMed:10801133) and V (PubMed:10801133). A shorter protein beta* may be important for increasing survival after UV irradiation, and stimulates DNA synthesis with increased processivity in the presence of core Pol III plus the clamp loader complex (PubMed:8576210, PubMed:8576212)." /translation="MKFTVEREHLLKPLQQVSGPLGGRPTLPILGNLLLQVADGTLSLTGTDLEMEMVARVALVQPHEPGATTVPARKFFDICRGLPEGAEIAVQLEGERMLVRSGRSRFSLSTLPAADFPNLDDWQSEVEFTLPQATMKRLIEATQFSMAHQDVRYYLNGMLFETEGEELRTVATDGHRLAVCSMPIGQSLPSHSVIVPRKGVIELMRMLDGGDNPLRVQIGSNNIRAHVGDFIFTSKLVDGRFPDYRRVLPKNPDKHLEAGCDLLKQAFARAAILSNEKFRGVRLYVSENQLKITANNPEQEEAEEILDVTYSGAEMEIGFNVSYVLDVLNALKCENVRMMLTDSVSSVQIEDAASQSAAYVVMPMRL" 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 3165..3224 /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 TGAAATTCAC 2041 CGTCGAACGT GAACACCTAC TAAAACCGCT ACAGCAGGTC TCAGGACCGC TAGGAGGACG 2101 TCCGACCCTA CCGATCCTAG GAAACCTACT ACTACAGGTC GCAGACGGAA CCCTATCACT 2161 AACCGGAACC GACCTAGAAA TGGAAATGGT CGCACGTGTC GCACTAGTCC AGCCGCACGA 2221 ACCGGGAGCA ACCACCGTCC CGGCACGTAA ATTCTTCGAC ATCTGCCGTG GACTACCGGA 2281 AGGAGCAGAA ATCGCAGTCC AGCTAGAAGG AGAACGTATG CTAGTCCGTT CAGGACGTTC 2341 ACGTTTCTCA CTATCAACCC TACCGGCAGC AGACTTCCCG AACCTAGACG ACTGGCAGTC 2401 AGAAGTCGAA TTCACCCTAC CGCAGGCAAC CATGAAACGT CTAATCGAAG CAACCCAGTT 2461 CTCAATGGCA CACCAGGACG TCCGTTACTA CCTAAACGGA ATGCTATTCG AAACCGAAGG 2521 AGAAGAACTA CGTACCGTCG CAACCGACGG ACACCGTCTA GCAGTCTGCT CAATGCCGAT 2581 CGGACAGTCA CTACCGTCAC ACTCAGTCAT CGTCCCGCGT AAAGGAGTCA TCGAACTAAT 2641 GCGTATGCTA GACGGAGGAG ACAACCCGCT ACGTGTCCAG ATCGGATCAA ACAACATCCG 2701 TGCACACGTC GGAGACTTCA TCTTCACCTC AAAACTAGTC GACGGACGTT TCCCGGACTA 2761 CCGTCGTGTC CTACCGAAAA ACCCGGACAA ACACCTAGAA GCAGGATGCG ACCTACTAAA 2821 ACAGGCATTC GCACGTGCAG CAATCCTATC AAACGAAAAA TTCCGTGGAG TCCGTCTATA 2881 CGTCTCAGAA AACCAGCTAA AAATCACCGC AAACAACCCG GAACAGGAAG AAGCAGAAGA 2941 AATCCTAGAC GTCACCTACT CAGGAGCAGA AATGGAAATC GGATTCAACG TCTCATACGT 3001 CCTAGACGTC CTAAACGCAC TAAAATGCGA AAACGTCCGT ATGATGCTAA CCGACTCAGT 3061 CTCATCAGTC CAGATCGAAG ACGCAGCATC ACAGTCAGCA GCATACGTCG TCATGCCGAT 3121 GCGTCTATGA AGAGCTTCGA GACCCCGTGC GACAAGATTT CAAGCCGCTC CCTCGGAATG 3181 TAAGGTAGGG ATAACAGGGT AATTACATAT ACCCTGAATC ATAG //