LOCUS       pEPQD0KN0244_(pJL1-NET_ 2372 bp ds-DNA     circular     16-OCT-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     primer_bind     456..473
                     /label="M13 Forward"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer_bind     457..473
                     /label="M13 fwd"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    723..829
                     /label="3' UTR and T7 terminator from pJL1"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     CDS             complement(847..1662)
                     /label="aminoglycoside O-phosphotransferase APH(3')-Ia, kanamycin resistance (kanR)"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    93..99
                     /label="RBS"
                     /ApEinfo_revcolor=#800080
                     /ApEinfo_fwdcolor=#800080
     primer_bind     complement(768..786)
                     /label="T7 Term"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    complement(810..824)
                     /label="T7 term hairpin R"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer_bind     complement(556..578)
                     /label="M13/pUC Reverse"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     complement(543..559)
                     /label="M13 rev"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer_bind     229..248
                     /label="pRS-marker"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    108..122
                     /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
     primer_bind     456..473
                     /label="M13-fwd"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    789..803
                     /label="T7 term hairpin F"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     RBS             78..100
                     /label="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    join(763..2372,1..24)
                     /label="backbone from pJL1-sfGFP (Addgene #69496)"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     terminator      782..829
                     /label="T7 terminator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer_bind     complement(894..921)
                     /label="QMD063 primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     promoter        28..46
                     /label="T7 promoter"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    complement(155..691)
                     /label="lacZ expression casette"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     protein_bind    636..657
                     /label="CAP binding site"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer_bind     complement(81..110)
                     /label="AN163\vector\amp\rev"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     rep_origin      1784..2372
                     /label="pUC origin of replication (derivative of pBR322/pMB1/ColE1)"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer_bind     1570..1589
                     /label="Kan-R"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     CDS             complement(224..547)
                     /label="lacZ beta-galactosidase alpha peptide"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer_bind     1654..1673
                     /label="pENTR-R"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    complement(554..677)
                     /label="Plac promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     protein_bind    567..583
                     /label="lac operator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    28..104
                     /label="T7 promoter and 5' UTR from pJL1"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        complement(591..621)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     442..464
                     /label="M13/pUC Forward"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer_bind     complement(543..559)
                     /label="M13 Reverse"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    474..530
                     /label="MCS"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer_bind     complement(539..559)
                     /label="M13-rev"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          complement(142..161)
                     /label="QMD129_MEKKIBsaI_lift_R, 61.3"
                     /note="sequence: ggcatccgcttacagacaag"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     primer          2306..2325
                     /label="QMD132_LETprom2_F, 61.4"
                     /note="sequence: gtttcgccacctctgacttg"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          complement(767..785)
                     /label="QMD061_T7_term"
                     /note="sequence: ctagttattgctcagcggt"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          124..147
                     /label="QMD378_24pJL1(26pEU)accept_to_CCAT_F"
                     /note="sequence: tctGAAGACtcCCATtGAGACCgtcacagcttgtc"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          2309..2327
                     /label="QMD062_pJL1_outside(BR322)_F"
                     /note="sequence: tcgccacctctgacttgag"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          2273..2296
                     /label="QMD131_LETprom1_F, 61.7, -3.89 self"
                     /note="sequence: gggaaacgcctggtatctttatag"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(925..948)
                     /label="QMD134_LETterm4_R, 61.2"
                     /note="sequence: ttttctccttcattacagaaacgg"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          complement(894..921)
                     /label="QMD063_pJL1_outside(kanR)_R"
                     /note="sequence: tttcaaaaatatggtattgataatcctg"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          complement(832..858)
                     /label="QMD133_LETterm3_R, 61.4"
                     /note="sequence: gagtttttctaatcagaattggctttc"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          complement(129..149)
                     /label="draft primer 61,6"
                     /note="sequence: cagacaagctgtgacGGTCTC"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          28..47
                     /label="QMD060_T7_prom"
                     /note="sequence: taatacgactcactataggg"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
ORIGIN
        1 aacgccagca acgcgatccc gcgaaattaa tacgactcac tatagggaca ccacaacggt
       61 ttccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg GAGAAAAAAA
      121 TATCCATtGA GACCgtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
      181 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
      241 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
      301 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
      361 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
      421 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcg
      481 agctcggtac ccggggatcc tctagagtcg acctgcaggc atgcaagctt ggcgtaatca
      541 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
      601 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
      661 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct aGGTCTCaGC
      721 TTcggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
      781 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaagccaa
      841 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
      901 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
      961 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     1021 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     1081 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
     1141 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     1201 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     1261 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     1321 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     1381 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     1441 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     1501 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     1561 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     1621 tcgcggcttc gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     1681 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     1741 acatcagaga ttttgagaca caacgtgaga tcaaaggatc ttcttgagat cctttttttc
     1801 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     1861 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     1921 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1981 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2041 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2101 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2161 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2221 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2281 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2341 gatgctcgtc aggggggcgg agcctatgga aa
//