LOCUS       pEPQDKN0329_(pJL1-NET-T 2529 bp ds-DNA     circular     16-OCT-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    complement(686..691)
                     /label="XbaI"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    1566..1580
                     /label="T7 term hairpin F"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    1500..1606
                     /label="3' UTR and T7 terminator from pJL1"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    648..724
                     /label="T7 promoter and 5' UTR from pJL1"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    728..742
                     /label="NET tag (N-terminal Expression Tag), MEKKI, from chloramphenicol acetyltransferase, minimizes secondary structure with RBS and 5' UTR"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     terminator      1559..1606
                     /label="T7 terminator"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    complement(1587..1601)
                     /label="T7 term hairpin R"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer_bind     557..575
                     /label="QMD062 primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     promoter        648..665
                     /label="T7 promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer_bind     complement(1544..1562)
                     /label="QMD061 primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    1460..1492
                     /label="HiBiT"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     RBS             698..720
                     /label="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    743..1453
                     /label="TEV protease S219V"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer_bind     648..667
                     /label="QMD060 primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     rep_origin      32..620
                     /label="pUC origin of replication (derivative of pBR322/pMB1/ColE1)"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    686..691
                     /label="XbaI"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer_bind     521..540
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     CDS             complement(1624..2439)
                     /label="aminoglycoside O-phosphotransferase APH(3')-Ia, kanamycin resistance (kanR)"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
ORIGIN
        1 TTGAGACACA ACGTGAGATC AAAGGATCTT CTTGAGATCC TTTTTTTCTG CGCGTAATCT
       61 GCTGCTTGCA AACAAAAAAA CCACCGCTAC CAGCGGTGGT TTGTTTGCCG GATCAAGAGC
      121 TACCAACTCT TTTTCCGAAG GTAACTGGCT TCAGCAGAGC GCAGATACCA AATACTGTTC
      181 TTCTAGTGTA GCCGTAGTTA GGCCACCACT TCAAGAACTC TGTAGCACCG CCTACATACC
      241 TCGCTCTGCT AATCCTGTTA CCAGTGGCTG CTGCCAGTGG CGATAAGTCG TGTCTTACCG
      301 GGTTGGACTC AAGACGATAG TTACCGGATA AGGCGCAGCG GTCGGGCTGA ACGGGGGGTT
      361 CGTGCACACA GCCCAGCTTG GAGCGAACGA CCTACACCGA ACTGAGATAC CTACAGCGTG
      421 AGCTATGAGA AAGCGCCACG CTTCCCGAAG GGAGAAAGGC GGACAGGTAT CCGGTAAGCG
      481 GCAGGGTCGG AACAGGAGAG CGCACGAGGG AGCTTCCAGG GGGAAACGCC TGGTATCTTT
      541 ATAGTCCTGT CGGGTTTCGC CACCTCTGAC TTGAGCGTCG ATTTTTGTGA TGCTCGTCAG
      601 GGGGGCGGAG CCTATGGAAA AACGCCAGCA ACGCGATCCC GCGAAATTAA TACGACTCAC
      661 TATAGGGACA CCACAACGGT TTCCCTCTAG AAATAATTTT GTTTAACTTT AAGAAGGAGA
      721 TATACATATG GAGAAAAAAA TAATGGGTGA GTCTCTGTTC AAAGGCCCAC GCGACTATAA
      781 TCCCATCTCC TCGACGATCT GCCACTTAAC CAACGAGTCG GACGGACATA CGACCAGTCT
      841 GTACGGCATC GGGTTCGGCC CGTTTATTAT CACTAATAAA CATCTGTTCC GTCGCAACAA
      901 CGGTACCTTA CTTGTACAGT CTCTTCACGG CGTGTTTAAA GTGAAGAATA CTACTACCCT
      961 TCAGCAGCAT CTGATCGACG GTCGCGATAT GATTATCATC CGTATGCCGA AAGACTTTCC
     1021 GCCCTTCCCG CAGAAATTAA AGTTCCGCGA ACCCCAGCGC GAGGAACGTA TTTGCTTGGT
     1081 TACCACAAAT TTTCAGACAA AATCAATGAG CTCAATGGTT AGCGATACGA GCTGTACTTT
     1141 TCCGAGCAGT GACGGTATCT TTTGGAAACA CTGGATCCAG ACGAAAGACG GCCAATGCGG
     1201 ATCACCGCTG GTGAGTACCC GTGACGGCTT TATCGTGGGC ATTCATTCCG CCAGCAACTT
     1261 TACTAATACC AATAACTACT TTACCTCAGT ACCCAAGAAT TTTATGGAGC TGTTAACCAA
     1321 CCAAGAAGCA CAACAATGGG TGAGCGGGTG GCGTCTGAAC GCCGATAGCG TTCTGTGGGG
     1381 TGGTCACAAG GTGTTTATGG TCAAGCCTGA GGAACCGTTC CAACCTGTGA AAGAGGCCAC
     1441 CCAGTTGATG AACGGTTCGG TGAGCGGCTG GCGGCTGTTC AAGAAGATTA GCTGAGCTTC
     1501 GGCTGCTAAC AAAGCCCGAA AGGAAGCTGA GTTGGCTGCT GCCACCGCTG AGCAATAACT
     1561 AGCATAACCC CTTGGGGCCT CTAAACGGGT CTTGAGGGGT TTTTTGCTGA AAGCCAATTC
     1621 TGATTAGAAA AACTCATCGA GCATCAAATG AAACTGCAAT TTATTCATAT CAGGATTATC
     1681 AATACCATAT TTTTGAAAAA GCCGTTTCTG TAATGAAGGA GAAAACTCAC CGAGGCAGTT
     1741 CCATAGGATG GCAAGATCCT GGTATCGGTC TGCGATTCCG ACTCGTCCAA CATCAATACA
     1801 ACCTATTAAT TTCCCCTCGT CAAAAATAAG GTTATCAAGT GAGAAATCAC CATGAGTGAC
     1861 GACTGAATCC GGTGAGAATG GCAAAAGCTT ATGCATTTCT TTCCAGACTT GTTCAACAGG
     1921 CCAGCCATTA CGCTCGTCAT CAAAATCACT CGCATCAACC AAACCGTTAT TCATTCGTGA
     1981 TTGCGCCTGA GCGAGACGAA ATACGCGATC GCTGTTAAAA GGACAATTAC AAACAGGAAT
     2041 CGAATGCAAC CGGCGCAGGA ACACTGCCAG CGCATCAACA ATATTTTCAC CTGAATCAGG
     2101 ATATTCTTCT AATACCTGGA ATGCTGTTTT CCCGGGGATC GCAGTGGTGA GTAACCATGC
     2161 ATCATCAGGA GTACGGATAA AATGCTTGAT GGTCGGAAGA GGCATAAATT CCGTCAGCCA
     2221 GTTTAGTCTG ACCATCTCAT CTGTAACATC ATTGGCAACG CTACCTTTGC CATGTTTCAG
     2281 AAACAACTCT GGCGCATCGG GCTTCCCATA CAATCGATAG ATTGTCGCAC CTGATTGCCC
     2341 GACATTATCG CGAGCCCATT TATACCCATA TAAATCAGCA TCCATGTTGG AATTTAATCG
     2401 CGGCTTCGAG CAAGACGTTT CCCGTTGAAT ATGGCTCATA ACACCCCTTG TATTACTGTT
     2461 TATGTAAGCA GACAGTTTTA TTGTTCATGA TGATATATTT TTATCTTGTG CAATGTAACA
     2521 TCAGAGATT
//