LOCUS       Exported     3128 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..3128
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
 polymerase"
     terminator      1165..1212
                     /note="transcription terminator for bacteriophage T7 RNA polymerase"
                     /label="T7 terminator"
     primer_bind     3060..3076
                     /label="M13 for"
                     /note="common sequencing primer, one of multiple similar variants"
     terminator      3077..3108
                     /label="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription terminator"
     CDS             29..1033
                     /label="holB"
                     /note="Part of the beta sliding clamp loading complex, which hydrolyzes ATP to load the beta clamp onto primed DNA to form the DNA replication pre-initiation complex (PubMed:2040637). DNA polymerase III is a complex, multichain enzyme responsible for most of the replicative synthesis in bacteria. This DNA polymerase also exhibits 3' to 5' exonuclease activity. The gamma complex (gamma(3),delta,delta') is thought to load beta dimers onto DNA by binding ATP which alters the complex's conformation so it can bind beta sliding clamp dimers and open them at one interface. Primed DNA is recognized, ATP is hydrolyzed releasing the gamma complex and closing the beta sliding clamp ring around the primed DNA (PubMed:9927437)."
                     /translation="MRWYPWLRPDFEKLVASYQAGRGHHALLIQALPGMGDDALIYALSRYLLCQQPQGHKSCGHCRGCQLMQAGTHPDYYTLAPEKGKNTLGVDAVREVTEKLNEHARLGGAKVVWVTDAALLTDAAANALLKTLEEPPAETWFFLATREPERLLATLRSRCRLHYLAPPPEQYAVTWLSREVTMSQDALLAALRLSAGSPGAALALFQGDNWQARETLCQALAYSVPSGDWYSLLAALNHEQAPARLHWLATLLMDALKRHHGAAQVTNVDVPGLVAELANHLSPSRLQAILGDVCHIREQLMSVTGINRELLITDLLLRIEHYLQPGVVLPVPHL"
     primer_bind     complement(1217..1233)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar variants"
     rep_origin      complement(1300..1888)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     CDS             complement(2079..2939)
                     /label="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     promoter        complement(2940..3044)
                     /label="bla promoter"
                     /note="ampicillin resistance gene promoter"
     misc_feature    1068..1127
                     /label="FreeGenes Barcode"
                     /note="FreeGenes Barcode. Defined as I-SceI site flanked by 21 base pair random sequences."
ORIGIN
        1 AAACCGGAGC CATACAGTAC GGTCTCTAAT GCGTTGGTAC CCGTGGCTAC GTCCGGACTT
       61 CGAAAAACTA GTCGCATCAT ACCAGGCAGG ACGTGGACAC CACGCACTAC TAATCCAGGC
      121 ACTACCGGGA ATGGGAGACG ACGCACTAAT CTACGCACTA TCACGTTACC TACTATGCCA
      181 GCAGCCGCAG GGACACAAAT CATGCGGACA CTGCCGTGGA TGCCAGCTAA TGCAGGCAGG
      241 AACCCACCCG GACTACTACA CCCTAGCACC GGAAAAAGGA AAAAACACCC TAGGAGTCGA
      301 CGCAGTCCGT GAAGTCACCG AAAAACTAAA CGAACACGCA CGTCTAGGAG GAGCAAAAGT
      361 CGTCTGGGTC ACCGACGCAG CACTACTAAC CGACGCAGCA GCAAACGCAC TACTAAAAAC
      421 CCTAGAAGAA CCGCCGGCAG AAACCTGGTT CTTCCTAGCA ACCCGTGAAC CGGAACGTCT
      481 ACTAGCAACC CTACGTTCAC GTTGCCGTCT ACACTACCTA GCACCGCCGC CGGAACAGTA
      541 CGCAGTCACC TGGCTATCAC GTGAAGTCAC CATGTCACAG GACGCACTAC TAGCAGCACT
      601 ACGTCTATCA GCAGGATCAC CGGGAGCAGC ACTAGCACTA TTCCAGGGAG ACAACTGGCA
      661 GGCACGTGAA ACCCTATGCC AGGCACTAGC ATACTCAGTC CCGTCAGGAG ACTGGTACTC
      721 ACTACTAGCA GCACTAAACC ACGAACAGGC ACCGGCACGT CTACACTGGC TAGCAACCCT
      781 ACTAATGGAC GCACTAAAAC GTCACCACGG AGCAGCACAG GTCACCAACG TCGACGTCCC
      841 GGGACTAGTC GCAGAACTAG CAAACCACCT ATCACCGTCA CGTCTACAGG CAATCCTAGG
      901 AGACGTCTGC CACATCCGTG AACAGCTAAT GTCAGTCACC GGAATCAACC GTGAACTACT
      961 AATCACCGAC CTACTACTAC GTATCGAACA CTACCTACAG CCGGGAGTCG TCCTACCGGT
     1021 CCCGCACCTA TGAAGAGCTT CGAGACCACA GGAAGCAAGG TATACGCCTC CCTCTCGGAA
     1081 CGCATATTTA GGGATAACAG GGTAATCCAT TGGACAGGCG AGCAGGACGC TGAGGTGTCA
     1141 ATCGTCGGAG CCGCTGAGCA ATAACTAGCA TAACCCCTTG GGGCCTCTAA ACGGGTCTTG
     1201 AGGGGTTTTT TGCATGGTCA TAGCTGTTTC CTGAGAGCTT GGCAGGTGAT GACACACATT
     1261 AACAAATTTC GTGAGGAGTC TCCAGAAGAA TGCCATTAAT TTCCATAGGC TCCGCCCCCC
     1321 TGACGAGCAT CACAAAAATC GACGCTCAAG TCAGAGGTGG CGAAACCCGA CAGGACTATA
     1381 AAGATACCAG GCGTTTCCCC CTGGAAGCTC CCTCGTGCGC TCTCCTGTTC CGACCCTGCC
     1441 GCTTACCGGA TACCTGTCCG CCTTTCTCCC TTCGGGAAGC GTGGCGCTTT CTCATAGCTC
     1501 ACGCTGTAGG TATCTCAGTT CGGTGTAGGT CGTTCGCTCC AAGCTGGGCT GTGTGCACGA
     1561 ACCCCCCGTT CAGCCCGACC GCTGCGCCTT ATCCGGTAAC TATCGTCTTG AGTCCAACCC
     1621 GGTAAGACAC GACTTATCGC CACTGGCAGC AGCCACTGGT AACAGGATTA GCAGAGCGAG
     1681 GTATGTAGGC GGTGCTACAG AGTTCTTGAA GTGGTGGCCT AACTACGGCT ACACTAGAAG
     1741 AACAGTATTT GGTATCTGCG CTCTGCTGAA GCCAGTTACC TTCGGAAAAA GAGTTGGTAG
     1801 CTCTTGATCC GGCAAACAAA CCACCGCTGG TAGCGGTGGT TTTTTTGTTT GCAAGCAGCA
     1861 GATTACGCGC AGAAAAAAAG GATCTCAAGA AGGCCTACTA TTAGCAACAA CGATCCTTTG
     1921 ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT CACGTTAAGG GATTTTGGTC
     1981 ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTAA ATTAAAAATG AAGTTTTAAA
     2041 TCAATCTAAA GTATATATGA GTAAACTTGG TCTGACAGTT ACCAATGCTT AATCAGTGAG
     2101 GCACCTATCT CAGCGATCTG TCTATTTCGT TCATCCATAG TTGCCTGACT CCCCGTCGTG
     2161 TAGATAACTA CGATACGGGA GGGCTTACCA TCTGGCCCCA GTGCTGCAAT GATACCGCGA
     2221 GAACCACGCT CACCGGCTCC AGATTTATCA GCAATAAACC AGCCAGCCGG AAGGGCCGAG
     2281 CGCAGAAGTG GTCCTGCAAC TTTATCCGCC TCCATCCAGT CTATTAATTG TTGCCGGGAA
     2341 GCTAGAGTAA GTAGTTCGCC AGTTAATAGT TTGCGCAACG TTGTTGCCAT TGCTACAGGC
     2401 ATCGTGGTGT CACGCTCGTC GTTTGGTATG GCTTCATTCA GCTCCGGTTC CCAACGATCA
     2461 AGGCGAGTTA CATGATCCCC CATGTTGTGC AAAAAAGCGG TTAGCTCCTT CGGTCCTCCG
     2521 ATCGTTGTCA GAAGTAAGTT GGCCGCAGTG TTATCACTCA TGGTTATGGC AGCACTGCAT
     2581 AATTCTCTTA CTGTCATGCC ATCCGTAAGA TGCTTTTCTG TGACTGGTGA GTACTCAACC
     2641 AAGTCATTCT GAGAATAGTG TATGCGGCGA CCGAGTTGCT CTTGCCCGGC GTCAATACGG
     2701 GATAATACCG CGCCACATAG CAGAACTTTA AAAGTGCTCA TCATTGGAAA ACGTTCTTCG
     2761 GGGCGAAAAC TCTCAAGGAT CTTACCGCTG TTGAGATCCA GTTCGATGTA ACCCACTCGT
     2821 GCACCCAACT GATCTTCAGC ATCTTTTACT TTCACCAGCG TTTCTGGGTG AGCAAAAACA
     2881 GGAAGGCAAA ATGCCGCAAA AAAGGGAATA AGGGCGACAC GGAAATGTTG AATACTCATA
     2941 CTCTTCCTTT TTCAATATTA TTGAAGCATT TATCAGGGTT ATTGTCTCAT GAGCGGATAC
     3001 ATATTTGAAT GTATTTAGAA AAATAAACAA ATAGGGGTTC CGCGCACCTG CACCAGTCAG
     3061 TAAAACGACG GCCAGTAGTC AAAAGCCTCC GACCGGAGGC TTTTGACTTG GTTCAGGTGG
     3121 AGTGGGAG
//