LOCUS       pR0219_His6-TwinStrep-S 9564 bp ds-DNA     circular     26-JUL-2018
DEFINITION  .
SOURCE      synthetic DNA construct                                            
  ORGANISM  recombinant plasmid                                                
FEATURES             Location/Qualifiers
     misc_feature    7525..7559
                     /label="F995 Primer"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     rep_origin      complement(1867..2549)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    4554..8042
                     /label="LbuC2c2"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    8052..8073
                     /label="R Primer 2"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_binding    4041..4063
                     /label="LacO"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    2688..2992
                     /label="GibsonInsert2"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    6757..6788
                     /label="F primer 2"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     CDS             complement(2647..3306)
                     /label="AmpR"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    5408..5438
                     /label="F primer 1"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    4113..4261
                     /label="6x His tag/ Twin Strep"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    6794..6817
                     /label="R primer 1"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    5763..5780
                     /label="R405 Primer"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    2785..2785
                     /label="replaced to remove BsaI site"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    4262..4553
                     /label="Sumo"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    5746..5773
                     /label="D401 Primer"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer_bind     4022..4041
                     /label="T7"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    5966..5987
                     /label="H473 Primer"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    5753..5777
                     /label="T403 Primer"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    7530..7567
                     /label="N997 Primer"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
ORIGIN
        1 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
       61 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
      121 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
      181 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
      241 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
      301 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
      361 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
      421 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat
      481 cactggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt
      541 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc
      601 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc
      661 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct
      721 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc
      781 agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta
      841 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga
      901 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc
      961 cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc
     1021 tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc
     1081 tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta
     1141 ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc
     1201 ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca
     1261 tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca
     1321 ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa
     1381 tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac
     1441 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     1501 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     1561 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
     1621 agcagattgt actgagagtg caccatatat gcggtgtgaa ataccgcaca gatgcgtaag
     1681 gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     1741 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     1801 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1861 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1921 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1981 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2041 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2101 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2161 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2221 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2281 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     2341 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     2401 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2461 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     2521 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     2581 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     2641 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     2701 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     2761 ccccagtgct gcaatgatac cgcgtgaccc acgctcaccg gctccagatt tatcagcaat
     2821 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     2881 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     2941 caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     3001 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     3061 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     3121 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     3181 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     3241 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     3301 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     3361 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     3421 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     3481 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     3541 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     3601 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     3661 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aaccgggatc
     3721 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     3781 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     3841 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     3901 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     3961 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
     4021 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
     4081 ttgtttaact ttaagaagga gatataccat gggcagcagc catcatcatc atcatcacag
     4141 cagcggcctg gtgccgcgcg gcagccatat ggctagctgg agccatccgc agtttgaaaa
     4201 aggtggtggt agcggtggtg gttcaggtgg tagtgcatgg tcacaccctc agtttgagaa
     4261 aatgtcggac tcagaagtca atcaagaagc taagccagag gtcaagccag aagtcaagcc
     4321 tgagactcac atcaatttaa aggtgtccga tggatcttca gagatcttct tcaagatcaa
     4381 aaagaccact cctttaagaa ggctgatgga agcgttcgct aaaagacagg gtaaggaaat
     4441 ggactcctta agattcttgt acgacggtat tagaatccaa gctgatcaga cccctgaaga
     4501 tttggacatg gaggataacg atattattga ggctcacaga gaacagattg gtgGATCTAT
     4561 GAAGGTGACA AAAGTGGGCG GCATCAGCCA CAAGAAGTAT ACCTCCGAGG GCAGGCTGGT
     4621 GAAGTCTGAG AGCGAGGAGA ACAGGACCGA CGAGCGCCTG TCCGCCCTGC TGAATATGAG
     4681 GCTGGATATG TATATCAAGA ACCCCAGCTC CACCGAGACA AAGGAGAATC AGAAGCGCAT
     4741 CGGCAAGCTG AAGAAGTTCT TTTCCAACAA GATGGTGTAT CTGAAGGACA ATACACTGTC
     4801 TCTGAAGAAT GGCAAGAAGG AGAACATCGA CCGGGAGTAC TCTGAGACAG ATATCCTGGA
     4861 GAGCGACGTG AGAGATAAGA AGAACTTCGC CGTGCTGAAG AAGATCTACC TGAATGAGAA
     4921 CGTGAACAGC GAGGAGCTGG AGGTGTTCCG GAATGATATC AAGAAGAAGC TGAACAAGAT
     4981 CAATTCTCTG AAGTACAGCT TTGAGAAGAA CAAGGCCAAT TATCAGAAGA TCAACGAGAA
     5041 CAATATCGAG AAGGTGGAGG GCAAGAGCAA GAGAAATATC ATCTACGACT ACTATAGGGA
     5101 GTCCGCCAAG CGGGACGCCT ACGTGAGCAA CGTGAAGGAG GCCTTTGACA AGCTGTATAA
     5161 GGAGGAGGAT ATCGCCAAGC TGGTGCTGGA GATCGAGAAT CTGACCAAGC TGGAGAAGTA
     5221 CAAGATCAGG GAGTTCTATC ACGAGATCAT CGGCCGCAAG AATGACAAGG AGAACTTTGC
     5281 CAAGATCATC TACGAGGAGA TCCAGAACGT GAACAATATG AAGGAGCTGA TCGAGAAGGT
     5341 GCCTGATATG TCCGAGCTGA AGAAGTCTCA GGTGTTCTAT AAGTACTATC TGGACAAGGA
     5401 GGAGCTGAAC GATAAGAATA TCAAGTACGC CTTCTGCCAC TTTGTGGAGA TCGAGATGAG
     5461 CCAGCTGCTG AAGAACTACG TGTATAAGAG GCTGTCCAAC ATCTCTAATG ACAAGATCAA
     5521 GCGCATCTTT GAGTATCAGA ATCTGAAGAA GCTGATCGAG AATAAGCTGC TGAACAAGCT
     5581 GGATACATAC GTGAGAAATT GTGGCAAGTA TAACTACTAT CTGCAGGACG GCGAGATCGC
     5641 CACCTCCGAT TTCATCGCCC GGAACAGACA GAATGAGGCC TTTCTGCGGA ACATCATCGG
     5701 CGTGTCTAGC GTGGCCTACT TCTCTCTGAG AAATATCCTG GAGACAGAAA ACGAGAATGA
     5761 CATCACAGGC AGGATGCGCG GCAAGACCGT GAAGAACAAT AAGGGCGAGG AGAAGTACGT
     5821 GTCTGGCGAG GTGGATAAGA TCTACAACGA GAACAAGAAG AATGAGGTGA AGGAGAACCT
     5881 GAAGATGTTC TACAGCTATG ACTTTAACAT GGATAACAAG AATGAGATCG AGGACTTCTT
     5941 TGCCAATATC GATGAGGCCA TCTCCTCTAT CAGGCACGGC ATCGTGCACT TCAACCTGGA
     6001 GCTGGAGGGC AAGGACATCT TCGCCTTTAA GAATATCGCC CCAAGCGAGA TCTCCAAGAA
     6061 GATGTTTCAG AACGAGATCA ATGAGAAGAA GCTGAAGCTG AAGATCTTCC GGCAGCTGAA
     6121 CTCCGCCAAC GTGTTCAGAT ACCTGGAGAA GTATAAGATC CTGAACTACC TGAAGCGGAC
     6181 AAGATTCGAG TTTGTGAACA AGAATATCCC CTTCGTGCCT TCTTTTACCA AGCTGTATAG
     6241 CCGCATCGAC GATCTGAAGA ACTCTCTGGG CATCTACTGG AAAACCCCTA AGACAAACGA
     6301 CGATAATAAG ACAAAGGAGA TCATCGACGC CCAGATCTAT CTGCTGAAGA ACATCTACTA
     6361 TGGCGAGTTC CTGAATTACT TTATGTCTAA CAATGGCAAC TTCTTTGAGA TCAGCAAGGA
     6421 GATCATCGAG CTGAACAAGA ATGACAAGAG GAACCTGAAA ACCGGCTTCT ACAAGCTGCA
     6481 GAAGTTTGAG GATATCCAGG AGAAGATCCC AAAGGAGTAC CTGGCCAATA TCCAGTCCCT
     6541 GTATATGATC AACGCCGGCA ATCAGGACGA GGAGGAGAAG GACACATACA TCGATTTCAT
     6601 CCAGAAGATC TTCCTGAAGG GCTTTATGAC CTACCTGGCC AACAATGGCC GCCTGTCCCT
     6661 GATCTATATC GGCTCTGACG AGGAGACAAA CACAAGCCTG GCCGAGAAGA AGCAGGAGTT
     6721 CGATAAGTTT CTGAAGAAGT ACGAGCAGAA CAATAACATC AAGATCCCCT ATGAGATCAA
     6781 TGAGTTCCTG AGGGAGATCA AGCTGGGCAA CATCCTGAAG TACACAGAGC GCCTGAATAT
     6841 GTTTTATCTG ATCCTGAAGC TGCTGAATCA CAAGGAGCTG ACCAACCTGA AGGGCAGCCT
     6901 GGAGAAGTAC CAGTCCGCCA ACAAGGAGGA GGCCTTCTCC GACCAGCTGG AGCTGATCAA
     6961 CCTGCTGAAT CTGGATAATA ACAGGGTGAC AGAGGACTTT GAGCTGGAGG CCGATGAGAT
     7021 CGGCAAGTTC CTGGACTTTA ACGGCAATAA GGTGAAGGAT AACAAGGAGC TGAAGAAGTT
     7081 CGACACCAAT AAGATCTACT TTGATGGCGA GAACATCATC AAGCACCGCG CCTTCTACAA
     7141 TATCAAGAAG TATGGCATGC TGAACCTGCT GGAGAAGATC GCCGACAAGG CCGGCTACAA
     7201 GATCAGCATC GAGGAGCTGA AGAAGTATTC CAACAAGAAG AATGAGATCG AGAAGAATCA
     7261 CAAGATGCAG GAGAACCTGC ACCGGAAGTA TGCCAGGCCA CGCAAGGATG AGAAGTTCAC
     7321 AGACGAGGAT TACGAGAGCT ATAAGCAGGC CATCGAGAAC ATCGAGGAGT ACACCCACCT
     7381 GAAGAATAAG GTGGAGTTTA ACGAGCTGAA TCTGCTGCAG GGCCTGCTGC TGAGAATCCT
     7441 GCACAGACTG GTGGGCTACA CATCCATCTG GGAGCGGGAC CTGCGGTTCA GACTGAAGGG
     7501 CGAGTTTCCC GAGAACCAGT ATATCGAGGA GATCTTCAAC TTTGAGAATA AGAAGAACGT
     7561 GAAGTACAAG GGCGGCCAGA TCGTGGAGAA GTACATCAAG TTCTATAAGG AGCTGCACCA
     7621 GAATGACGAG GTGAAGATCA ACAAGTATAG CTCCGCCAAT ATCAAGGTGC TGAAGCAGGA
     7681 GAAGAAGGAT CTGTATATCC GGAACTATAT CGCCCACTTT AATTACATCC CTCACGCCGA
     7741 GATCAGCCTG CTGGAGGTGC TGGAGAACCT GCGGAAGCTG CTGAGCTATG ACAGAAAGCT
     7801 GAAGAATGCC GTGATGAAGT CCGTGGTGGA TATCCTGAAG GAGTACGGCT TCGTGGCCAC
     7861 ATTTAAGATC GGCGCCGACA AGAAGATCGG CATCCAGACC CTGGAGTCTG AGAAGATCGT
     7921 GCACCTGAAG AACCTGAAGA AGAAGAAGCT GATGACCGAT AGAAATAGCG AGGAGCTGTG
     7981 CAAGCTGGTG AAGATCATGT TCGAgtacaa gatggaggag aagaagtccg agaactaagg
     8041 ccgcactcga ggcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag
     8101 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     8161 tatccggata tcccgcaaga ggcccggcag taccggcata accaagccta tgcctacagc
     8221 atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca tacacggtgc
     8281 ctgactgcgt tagcaattta actgtgataa actaccgcat taaagcttat cgatgataag
     8341 ctgtcaaaca tgagaaacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
     8401 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
     8461 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
     8521 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
     8581 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
     8641 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
     8701 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
     8761 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
     8821 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
     8881 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
     8941 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
     9001 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
     9061 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
     9121 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
     9181 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
     9241 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
     9301 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
     9361 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
     9421 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     9481 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
     9541 gctcactcat taggcaccgg gatc
//