LOCUS       pEPQDKN0729_(pJL1-NET-T 2522 bp ds-DNA     circular     16-OCT-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    1560..1574
                     /label="T7 term hairpin F"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    729..743
                     /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      1553..1600
                     /label="T7 terminator"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    complement(1581..1595)
                     /label="T7 term hairpin R"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    1461..1489
                     /label="stop codon part from pEPQD0CM0030"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    747..1454
                     /label="TEV protease S219V (codon optimized for E coli by Twist)"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     promoter        649..666
                     /label="T7 promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer_bind     522..541
                     /label="pBR322ori-F"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     terminator      1558..1601
                     /label="T7\term"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer_bind     complement(702..731)
                     /label="AN163\vector\amp\rev"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    558..576
                     /label="QMD062 primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     RBS             699..721
                     /label="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    1461..1463
                     /label="stop codon"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer_bind     2341..2360
                     /label="Kan-R"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     CDS             complement(1618..2433)
                     /label="aminoglycoside O-phosphotransferase APH(3')-Ia, kanamycin resistance (kanR)"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    649..728
                     /label="T7 promoter and 5' UTR from pJL1"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer_bind     complement(1539..1557)
                     /label="T7 Term"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     rep_origin      33..621
                     /label="pUC origin of replication (derivative of pBR322/pMB1/ColE1)"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    complement(1538..1556)
                     /label="QMD061 primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer_bind     2425..2444
                     /label="pENTR-R"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    714..720
                     /label="RBS"
                     /ApEinfo_revcolor=#800080
                     /ApEinfo_fwdcolor=#800080
     misc_feature    1494..1600
                     /label="3' UTR and T7 terminator from pJL1"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
ORIGIN
        1 tttgagacac aacgtgagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
       61 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
      121 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
      181 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
      241 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
      301 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
      361 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
      421 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
      481 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
      541 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
      601 ggggggcgga gcctatggaa aaacgccagc aacgcgatcc cgcgaaatta atacgactca
      661 ctatagggac accacaacgg tttccctcta gaaataattt tgtttaactt taagaaggag
      721 atatacatat gGAGAAAAAA ATAATGGGTG AGTCTCTGTT CAAAGGCCCA CGCGACTATA
      781 ATCCCATCTC CTCGACGATC TGCCACTTAA CCAACGAGTC GGACGGACAT ACGACCAGTC
      841 TGTACGGCAT CGGGTTCGGC CCGTTTATTA TCACTAATAA ACATCTGTTC CGTCGCAACA
      901 ACGGTACCTT ACTTGTACAG TCTCTTCACG GCGTGTTTAA AGTGAAGAAT ACTACTACCC
      961 TTCAGCAGCA TCTGATCGAC GGTCGCGATA TGATTATCAT CCGTATGCCG AAAGACTTTC
     1021 CGCCCTTCCC GCAGAAATTA AAGTTCCGCG AACCCCAGCG CGAGGAACGT ATTTGCTTGG
     1081 TTACCACAAA TTTTCAGACA AAATCAATGA GCTCAATGGT TAGCGATACG AGCTGTACTT
     1141 TTCCGAGCAG TGACGGTATC TTTTGGAAAC ACTGGATCCA GACGAAAGAC GGCCAATGCG
     1201 GATCACCGCT GGTGAGTACC CGTGACGGCT TTATCGTGGG CATTCATTCC GCCAGCAACT
     1261 TTACTAATAC CAATAACTAC TTTACCTCAG TACCCAAGAA TTTTATGGAG CTGTTAACCA
     1321 ACCAAGAAGC ACAACAATGG GTGAGCGGGT GGCGTCTGAA CGCCGATAGC GTTCTGTGGG
     1381 GTGGTCACAA GGTGTTTATG GTCAAGCCTG AGGAACCGTT CCAACCTGTG AAAGAGGCCA
     1441 CCCAGTTGAT GAACGGTTCG taagtcgacc ggctgctaac aaagcccgaG CTTcggctgc
     1501 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     1561 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaagcca attctgatta
     1621 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     1681 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     1741 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     1801 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     1861 atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc
     1921 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     1981 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     2041 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     2101 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     2161 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     2221 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     2281 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     2341 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggctt
     2401 cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     2461 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     2521 at
//