LOCUS       pET-22b(+)_6His_SpyCatc 6155 bp ds-DNA     circular     19-NOV-2020
DEFINITION  .
KEYWORDS    "creator:Novagen (EMD Millipore)" "marker:AmpR"
            "organism:Escherichia coli"
COMMENT     Bacterial vector that encodes a signal sequence for inducible
            expression of proteins in the periplasm.
FEATURES             Location/Qualifiers
     promoter        86..163
                     /label="lacI promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             164..1246
                     /label="lacI"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     CDS             2055..2246
                     /label="rop"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     rep_origin      complement(2676..3264)
                     /label="ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     CDS             complement(3435..4226)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     CDS             complement(4227..4295)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     promoter        complement(4296..4400)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     rep_origin      complement(4427..4882)
                     /label="f1 ori"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          4908..4929
                     /label="SEQ22R"
                     /note="sequence: CCTCCTTTCAGCAAAAAACCCC"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          4919..4939
                     /label="T7 term. pri."
                     /note="sequence: CAAAAAACCCCTCAAGACCCG"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     terminator      4919..4966
                     /label="T7 terminator"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             complement(5030..5050)
                     /label="Translation 5050-5030"
                     /translation="HHHHHH*"
     CDS             complement(5033..5050)
                     /label="6xHis"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(5033..5050)
                     /label="MEI11"
                     /note="sequence: aaaaaaaCTCGAGGATCAGAACGCGACCCACCACCACCACCACCAC"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          complement(5034..5056)
                     /label="Fwd2"
                     /note="sequence: CTCGAGCACCACCACCACCACCA"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          complement(5036..5056)
                     /label="CK42"
                     /note="sequence: CTCGAGCACCACCACCACCAC"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
     CDS             complement(5051..5056)
                     /label="Translation 5056-5051"
                     /translation="LE"
     primer          5057..5095
                     /label="pREA145"
                     /note="sequence: TGGTGGTGGTGGTGCTCGAGttattaatactgttcataataaatatccaggctatccag"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     CDS             complement(5057..5449)
                     /label="GRFT"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             complement(5057..5857)
                     /label="Translation 5857-5057"
                     /translation="MGSSHHHHHHTSENLYFQGAMVDTLSGLSSEQGQSGDMTIEEDSATHIKFSKRDEDGKELAGATMELRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNGKATKGDAHIGSSSGLVPRGSLTHRKFGGSGGSPFSGLSSIAVRSGSYLDAIIIDGVHHGGSGGNLSPTFTFGSGEYISNMTIRSGDYIDNISFETNMGRRFGPYGGSGGSANTLSNVKVIQINGSAGDYLDSLDIYYEQY**"
     misc_feature    complement(5063..5425)
                     /label="GRFT coding sec"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          complement(5405..5425)
                     /label="GO22"
                     /note="sequence: ggatccagcagcggtctggttccgcgtggtagcctgacccatcgcaaattc"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    complement(5426..5431)
                     /label="Trombin cleavage site"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    complement(5450..5455)
                     /label="Linker"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          5456..5479
                     /label="GO20"
                     /note="sequence: accacgcggaaccagaccgctgctggatccaatatgagcgtcacctttagttgc"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    complement(5456..5794)
                     /label="spycatcher"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(5769..5797)
                     /label="GO21"
                     /note="sequence: cacACTAGTgaaaacctgtattttcagggcgccatggttgataccttatcaggtttatcaag"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             complement(5801..5821)
                     /label="TEV site"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(5828..5845)
                     /label="His Tag"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          5831..5852
                     /label="GO23"
                     /note="sequence: ggcgccctgaaaatacaggttttcACTAGTgtggtgatgatgatggtggctgcta"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer          complement(5839..5887)
                     /label="pREA144"
                     /note="sequence: CTCTAGATTAACTTTAAGAAGGAGGGTACCatgggtagcagccaccatc"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          complement(5840..5859)
                     /label="GRFT_pet22b_F (67)"
                     /note="sequence: ataacaattcccctctagaATTAAAGAGGAGAAAGGTACgcggatccatgggtagcagccaccat"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    5858..5863
                     /label="Cut Site"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     RBS             5864..5869
                     /label="RBS"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    complement(5872..5880)
                     /label="epsilon"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     protein_bind    5889..5913
                     /label="lac operator"
                     /ApEinfo_revcolor="#31849b"
                     /ApEinfo_fwdcolor="#31849b"
     promoter        complement(5914..5932)
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(6076..6095)
                     /label="SEQFWD22"
                     /note="sequence: CATACCCACGCCGAAACAAG"
                     /ApEinfo_revcolor="#f7977a"
                     /ApEinfo_fwdcolor="#f7977a"
ORIGIN
        1 GCGGCGGCGG TGCTCAACGG CCTCAACCTA CTACTGGGCT GCTTCCTAAT GCAGGAGTCG
       61 CATAAGGGAG AGCGTCGAGA TCCCGGACAC CATCGAATGG CGCAAAACCT TTCGCGGTAT
      121 GGCATGATAG CGCCCGGAAG AGAGTCAATT CAGGGTGGTG AATGTGAAAC CAGTAACGTT
      181 ATACGATGTC GCAGAGTATG CCGGTGTCTC TTATCAGACC GTTTCCCGCG TGGTGAACCA
      241 GGCCAGCCAC GTTTCTGCGA AAACGCGGGA AAAAGTGGAA GCGGCGATGG CGGAGCTGAA
      301 TTACATTCCC AACCGCGTGG CACAACAACT GGCGGGCAAA CAGTCGTTGC TGATTGGCGT
      361 TGCCACCTCC AGTCTGGCCC TGCACGCGCC GTCGCAAATT GTCGCGGCGA TTAAATCTCG
      421 CGCCGATCAA CTGGGTGCCA GCGTGGTGGT GTCGATGGTA GAACGAAGCG GCGTCGAAGC
      481 CTGTAAAGCG GCGGTGCACA ATCTTCTCGC GCAACGCGTC AGTGGGCTGA TCATTAACTA
      541 TCCGCTGGAT GACCAGGATG CCATTGCTGT GGAAGCTGCC TGCACTAATG TTCCGGCGTT
      601 ATTTCTTGAT GTCTCTGACC AGACACCCAT CAACAGTATT ATTTTCTCCC ATGAAGACGG
      661 TACGCGACTG GGCGTGGAGC ATCTGGTCGC ATTGGGTCAC CAGCAAATCG CGCTGTTAGC
      721 GGGCCCATTA AGTTCTGTCT CGGCGCGTCT GCGTCTGGCT GGCTGGCATA AATATCTCAC
      781 TCGCAATCAA ATTCAGCCGA TAGCGGAACG GGAAGGCGAC TGGAGTGCCA TGTCCGGTTT
      841 TCAACAAACC ATGCAAATGC TGAATGAGGG CATCGTTCCC ACTGCGATGC TGGTTGCCAA
      901 CGATCAGATG GCGCTGGGCG CAATGCGCGC CATTACCGAG TCCGGGCTGC GCGTTGGTGC
      961 GGATATCTCG GTAGTGGGAT ACGACGATAC CGAAGACAGC TCATGTTATA TCCCGCCGTT
     1021 AACCACCATC AAACAGGATT TTCGCCTGCT GGGGCAAACC AGCGTGGACC GCTTGCTGCA
     1081 ACTCTCTCAG GGCCAGGCGG TGAAGGGCAA TCAGCTGTTG CCCGTCTCAC TGGTGAAAAG
     1141 AAAAACCACC CTGGCGCCCA ATACGCAAAC CGCCTCTCCC CGCGCGTTGG CCGATTCATT
     1201 AATGCAGCTG GCACGACAGG TTTCCCGACT GGAAAGCGGG CAGTGAGCGC AACGCAATTA
     1261 ATGTAAGTTA GCTCACTCAT TAGGCACCGG GATCTCGACC GATGCCCTTG AGAGCCTTCA
     1321 ACCCAGTCAG CTCCTTCCGG TGGGCGCGGG GCATGACTAT CGTCGCCGCA CTTATGACTG
     1381 TCTTCTTTAT CATGCAACTC GTAGGACAGG TGCCGGCAGC GCTCTGGGTC ATTTTCGGCG
     1441 AGGACCGCTT TCGCTGGAGC GCGACGATGA TCGGCCTGTC GCTTGCGGTA TTCGGAATCT
     1501 TGCACGCCCT CGCTCAAGCC TTCGTCACTG GTCCCGCCAC CAAACGTTTC GGCGAGAAGC
     1561 AGGCCATTAT CGCCGGCATG GCGGCCCCAC GGGTGCGCAT GATCGTGCTC CTGTCGTTGA
     1621 GGACCCGGCT AGGCTGGCGG GGTTGCCTTA CTGGTTAGCA GAATGAATCA CCGATACGCG
     1681 AGCGAACGTG AAGCGACTGC TGCTGCAAAA CGTCTGCGAC CTGAGCAACA ACATGAATGG
     1741 TCTTCGGTTT CCGTGTTTCG TAAAGTCTGG AAACGCGGAA GTCAGCGCCC TGCACCATTA
     1801 TGTTCCGGAT CTGCATCGCA GGATGCTGCT GGCTACCCTG TGGAACACCT ACATCTGTAT
     1861 TAACGAAGCG CTGGCATTGA CCCTGAGTGA TTTTTCTCTG GTCCCGCCGC ATCCATACCG
     1921 CCAGTTGTTT ACCCTCACAA CGTTCCAGTA ACCGGGCATG TTCATCATCA GTAACCCGTA
     1981 TCGTGAGCAT CCTCTCTCGT TTCATCGGTA TCATTACCCC CATGAACAGA AATCCCCCTT
     2041 ACACGGAGGC ATCAGTGACC AAACAGGAAA AAACCGCCCT TAACATGGCC CGCTTTATCA
     2101 GAAGCCAGAC ATTAACGCTT CTGGAGAAAC TCAACGAGCT GGACGCGGAT GAACAGGCAG
     2161 ACATCTGTGA ATCGCTTCAC GACCACGCTG ATGAGCTTTA CCGCAGCTGC CTCGCGCGTT
     2221 TCGGTGATGA CGGTGAAAAC CTCTGACACA TGCAGCTCCC GGAGACGGTC ACAGCTTGTC
     2281 TGTAAGCGGA TGCCGGGAGC AGACAAGCCC GTCAGGGCGC GTCAGCGGGT GTTGGCGGGT
     2341 GTCGGGGCGC AGCCATGACC CAGTCACGTA GCGATAGCGG AGTGTATACT GGCTTAACTA
     2401 TGCGGCATCA GAGCAGATTG TACTGAGAGT GCACCATATA TGCGGTGTGA AATACCGCAC
     2461 AGATGCGTAA GGAGAAAATA CCGCATCAGG CGCTCTTCCG CTTCCTCGCT CACTGACTCG
     2521 CTGCGCTCGG TCGTTCGGCT GCGGCGAGCG GTATCAGCTC ACTCAAAGGC GGTAATACGG
     2581 TTATCCACAG AATCAGGGGA TAACGCAGGA AAGAACATGT GAGCAAAAGG CCAGCAAAAG
     2641 GCCAGGAACC GTAAAAAGGC CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC
     2701 GAGCATCACA AAAATCGACG CTCAAGTCAG AGGTGGCGAA ACCCGACAGG ACTATAAAGA
     2761 TACCAGGCGT TTCCCCCTGG AAGCTCCCTC GTGCGCTCTC CTGTTCCGAC CCTGCCGCTT
     2821 ACCGGATACC TGTCCGCCTT TCTCCCTTCG GGAAGCGTGG CGCTTTCTCA TAGCTCACGC
     2881 TGTAGGTATC TCAGTTCGGT GTAGGTCGTT CGCTCCAAGC TGGGCTGTGT GCACGAACCC
     2941 CCCGTTCAGC CCGACCGCTG CGCCTTATCC GGTAACTATC GTCTTGAGTC CAACCCGGTA
     3001 AGACACGACT TATCGCCACT GGCAGCAGCC ACTGGTAACA GGATTAGCAG AGCGAGGTAT
     3061 GTAGGCGGTG CTACAGAGTT CTTGAAGTGG TGGCCTAACT ACGGCTACAC TAGAAGGACA
     3121 GTATTTGGTA TCTGCGCTCT GCTGAAGCCA GTTACCTTCG GAAAAAGAGT TGGTAGCTCT
     3181 TGATCCGGCA AACAAACCAC CGCTGGTAGC GGTGGTTTTT TTGTTTGCAA GCAGCAGATT
     3241 ACGCGCAGAA AAAAAGGATC TCAAGAAGAT CCTTTGATCT TTTCTACGGG GTCTGACGCT
     3301 CAGTGGAACG AAAACTCACG TTAAGGGATT TTGGTCATGA GATTATCAAA AAGGATCTTC
     3361 ACCTAGATCC TTTTAAATTA AAAATGAAGT TTTAAATCAA TCTAAAGTAT ATATGAGTAA
     3421 ACTTGGTCTG ACAGTTACCA ATGCTTAATC AGTGAGGCAC CTATCTCAGC GATCTGTCTA
     3481 TTTCGTTCAT CCATAGTTGC CTGACTCCCC GTCGTGTAGA TAACTACGAT ACGGGAGGGC
     3541 TTACCATCTG GCCCCAGTGC TGCAATGATA CCGCGAGACC CACGCTCACC GGCTCCAGAT
     3601 TTATCAGCAA TAAACCAGCC AGCCGGAAGG GCCGAGCGCA GAAGTGGTCC TGCAACTTTA
     3661 TCCGCCTCCA TCCAGTCTAT TAATTGTTGC CGGGAAGCTA GAGTAAGTAG TTCGCCAGTT
     3721 AATAGTTTGC GCAACGTTGT TGCCATTGCT GCAGGCATCG TGGTGTCACG CTCGTCGTTT
     3781 GGTATGGCTT CATTCAGCTC CGGTTCCCAA CGATCAAGGC GAGTTACATG ATCCCCCATG
     3841 TTGTGCAAAA AAGCGGTTAG CTCCTTCGGT CCTCCGATCG TTGTCAGAAG TAAGTTGGCC
     3901 GCAGTGTTAT CACTCATGGT TATGGCAGCA CTGCATAATT CTCTTACTGT CATGCCATCC
     3961 GTAAGATGCT TTTCTGTGAC TGGTGAGTAC TCAACCAAGT CATTCTGAGA ATAGTGTATG
     4021 CGGCGACCGA GTTGCTCTTG CCCGGCGTCA ATACGGGATA ATACCGCGCC ACATAGCAGA
     4081 ACTTTAAAAG TGCTCATCAT TGGAAAACGT TCTTCGGGGC GAAAACTCTC AAGGATCTTA
     4141 CCGCTGTTGA GATCCAGTTC GATGTAACCC ACTCGTGCAC CCAACTGATC TTCAGCATCT
     4201 TTTACTTTCA CCAGCGTTTC TGGGTGAGCA AAAACAGGAA GGCAAAATGC CGCAAAAAAG
     4261 GGAATAAGGG CGACACGGAA ATGTTGAATA CTCATACTCT TCCTTTTTCA ATATTATTGA
     4321 AGCATTTATC AGGGTTATTG TCTCATGAGC GGATACATAT TTGAATGTAT TTAGAAAAAT
     4381 AAACAAATAG GGGTTCCGCG CACATTTCCC CGAAAAGTGC CACCTGAAAT TGTAAACGTT
     4441 AATATTTTGT TAAAATTCGC GTTAAATTTT TGTTAAATCA GCTCATTTTT TAACCAATAG
     4501 GCCGAAATCG GCAAAATCCC TTATAAATCA AAAGAATAGA CCGAGATAGG GTTGAGTGTT
     4561 GTTCCAGTTT GGAACAAGAG TCCACTATTA AAGAACGTGG ACTCCAACGT CAAAGGGCGA
     4621 AAAACCGTCT ATCAGGGCGA TGGCCCACTA CGTGAACCAT CACCCTAATC AAGTTTTTTG
     4681 GGGTCGAGGT GCCGTAAAGC ACTAAATCGG AACCCTAAAG GGAGCCCCCG ATTTAGAGCT
     4741 TGACGGGGAA AGCCGGCGAA CGTGGCGAGA AAGGAAGGGA AGAAAGCGAA AGGAGCGGGC
     4801 GCTAGGGCGC TGGCAAGTGT AGCGGTCACG CTGCGCGTAA CCACCACACC CGCCGCGCTT
     4861 AATGCGCCGC TACAGGGCGC GTCCCATTCG CCAATCCGGA TATAGTTCCT CCTTTCAGCA
     4921 AAAAACCCCT CAAGACCCGT TTAGAGGCCC CAAGGGGTTA TGCTAGTTAT TGCTCAGCGG
     4981 TGGCAGCAGC CAACTCAGCT TCCTTTCGGG CTTTGTTAGC AGCCGGATCT CAGTGGTGGT
     5041 GGTGGTGGTG CTCGAGttat taatactgtt cataataaat atccaggcta tccagataat
     5101 cgcctgcgct accgttgatc tggatcacct tcacgttgct cagggtattg gcgctaccac
     5161 cactaccacc atacggaccg aagcggcgac ccatgttggt ctcgaagctg atgttgtcga
     5221 tatagtcgcc gctgcggatg gtcatgttgc taatgtactc gccgctgcca aaggtaaagg
     5281 tcgggctcag attaccaccg ctaccaccat gatgcacgcc atcgatgatg atggcatcca
     5341 ggtagctacc actgcgcact gcaatgctgc tcagaccgct aaacgggcta ccaccgctgc
     5401 caccgaattt gcgatgggtc aggctaccac gcggaaccag accgctgctg gatccaatat
     5461 gagcgtcacc tttagttgct ttgccattta cagtaacctg accttgctca ttaactgtaa
     5521 aggtaatagc agttgctacc tcataaccgt ctggtgctgc ggtttcgaca aatgtatatt
     5581 ttcctggata caggtagaaa tctttcactt gtccatctga aatccatgta ctaatagttt
     5641 taccagatga atcacgcaac tccatagttg caccagctaa ctctttgccg tcctcatcac
     5701 gttttgagaa tttaatatgg gtagcactat cttcttcaat tgtcatatca ccggactgac
     5761 cttgctcact tgataaacct gataaggtat caaccatggc gccctgaaaa tacaggtttt
     5821 cACTAGTgtg gtgatgatga tggtggctgc tacccatGGT ACCCTCCTTC TTAAAGTTAA
     5881 TCTAGAGGGG AATTGTTATC CGCTCACAAT TCCCCTATAG TGAGTCGTAT TAATTTCGCG
     5941 GGATCGAGAT CTCGATCCTC TACGCCGGAC GCATCGTGGC CGGCATCACC GGCGCCACAG
     6001 GTGCGGTTGC TGGCGCCTAT ATCGCCGACA TCACCGATGG GGAAGATCGG GCTCGCCACT
     6061 TCGGGCTCAT GAGCGCTTGT TTCGGCGTGG GTATGGTGGC AGGCCCCGTG GCCGGGGGAC
     6121 TGTTGGGCGC CATCTCCTTG CATGCACCAT TCCTT
//