LOCUS       pEB2-mKate2             3929 bp    DNA     circular UNA 27-DEC-2017
DEFINITION  pEB2 plasmid with mKate2.
ACCESSION   urn.local...d9-87ybf4n
VERSION     urn.local...d9-87ybf4n
KEYWORDS    .
SOURCE      Escherichia coli
  ORGANISM  Escherichia coli
            .
FEATURES             Location/Qualifiers
     promoter        1..144
                     /created_by="User"
                     /modified_by="User"
                     /label="proC"
     motif           63..68
                     /created_by="User"
                     /modified_by="User"
                     /label="-35 Box"
     motif           86..91
                     /created_by="User"
                     /label="-10 Box"
     motif           98
                     /created_by="User"
                     /label="TransStart"
     misc_feature    145..170
                     /created_by="User"
                     /modified_by="User"
                     /label="UArbs"
     CDS             171..866
                     /note="derived from TagRFP, Genbank accession EF606900"
                     /codon_start=1
                     /transl_table=11
                     /product="monomeric red fluorescent protein TagRFP-T"
                     /protein_id="ACD03281.1"
                     /db_xref="GI:187373121"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQT
                     MRIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYED
                     GGVLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEG
                     RTDMALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQH
                     EVAVARYCDLPSKLGHKLNGMDELYK"
                     /modified_by="User"
                     /label="mKate2"
     motif           264..266
                     /created_by="User"
                     /modified_by="User"
                     /label="R32G"
     motif           294..296
                     /created_by="User"
                     /modified_by="User"
                     /label="K42R"
     motif           303..305
                     /created_by="User"
                     /modified_by="User"
                     /label="V45A"
     motif           405..407
                     /created_by="User"
                     /modified_by="User"
                     /label="L79F"
     motif           447..449
                     /created_by="User"
                     /modified_by="User"
                     /label="I93V"
     motif           504..506
                     /created_by="User"
                     /modified_by="User"
                     /label="N112D"
     motif           513..515
                     /created_by="User"
                     /modified_by="User"
                     /label="I115L"
     motif           534..536
                     /created_by="User"
                     /modified_by="User"
                     /label="N122R"
     motif           561..563
                     /created_by="User"
                     /modified_by="User"
                     /label="S131P"
     motif           597..599
                     /created_by="User"
                     /modified_by="User"
                     /label="N143S"
     motif           606..608
                     /created_by="User"
                     /modified_by="User"
                     /label="M146T"
     motif           633..635
                     /created_by="User"
                     /modified_by="User"
                     /label="R155E"
     motif           639..641
                     /created_by="User"
                     /modified_by="User"
                     /label="H157R"
     motif           642..644
                     /created_by="User"
                     /modified_by="User"
                     /label="S158A"
     motif           645..647
                     /created_by="User"
                     /modified_by="User"
                     /label="Q159D"
     motif           675..677
                     /created_by="User"
                     /modified_by="User"
                     /label="Y169H"
     motif           681..683
                     /created_by="User"
                     /modified_by="User"
                     /label="H171I"
     motif           687..689
                     /created_by="User"
                     /modified_by="User"
                     /label="S173N"
     motif           690..692
                     /created_by="User"
                     /modified_by="User"
                     /label="F174L"
     motif           744..746
                     /created_by="User"
                     /modified_by="User"
                     /label="F192V"
     motif           747..749
                     /created_by="User"
                     /modified_by="User"
                     /label="H193Y"
     motif           750..752
                     /created_by="User"
                     /modified_by="User"
                     /label="F194Y"
     motif           759..761
                     /created_by="User"
                     /modified_by="User"
                     /label="H197R"
     motif           816..818
                     /created_by="User"
                     /modified_by="User"
                     /label="M216V"
     motif           828..830
                     /created_by="User"
                     /modified_by="User"
                     /label="K220R"
     terminator      898..956
                     /created_by="User"
                     /modified_by="User"
                     /label="rrnB T1 terminator"
     rep_origin      complement(957..2840)
                     /created_by="User"
                     /modified_by="User"
                     /label="Origin of Rep. from pSC101"
     repeat_region   complement(980..996)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     primer_bind     complement(1018..1041)
                     /Sequence="AAAGGGAAAACTGTCCATATGCAC"
                     /Hairpin_Tm="39.7"
                     /Tm="59.8"
                     /Self_Dimer_Tm="None"
                     /%GC="41.7"
                     /created_by="primer3"
                     /label="T1_from_mSC101_R02"
     repeat_region   complement(1098..1112)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     CDS             complement(1123..2073)
                     /note="unnamed protein product; rep101 protein"
                     /codon_start=1
                     /transl_table=11
                     /protein_id="CAA25276.1"
                     /db_xref="GI:42705"
                     /db_xref="GOA:Q47589"
                     /db_xref="InterPro:IPR000525"
                     /db_xref="InterPro:IPR011991"
                     /db_xref="UniProtKB/TrEMBL:Q47589"
                     /modified_by="User"
                     /label="RepA"
     regulatory      complement(2075..2080)
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Dalgarno sequence"
                     /label="Shine-Dalgarno sequence regulatory"
     repeat_region   complement(2075..2097)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2100..2121)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     regulatory      complement(2104..2109)
                     /regulatory_class="promoter"
                     /note="-10 region"
                     /label="-10 region regulatory"
     regulatory      complement(2126..2131)
                     /regulatory_class="promoter"
                     /note="-35 region"
                     /label="-35 region regulatory"
     repeat_region   complement(2721..2738)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2744..2767)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2778..2794)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2807..2820)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     terminator      complement(2846..2911)
                     /created_by="User"
                     /modified_by="User"
                     /label="t(o) terminator from pSS9"
     CDS             complement(2924..3718)
                     /gene="aph"
                     /codon_start=1
                     /transl_table=11
                     /product="Aminoglycoside 3'-phosphotransferase (APH)
                     kanamycin/neomycin resistance protein"
                     /protein_id="AJW29743.1"
                     /db_xref="GI:768968340"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
                     /modified_by="User"
                     /label="Aph (KmR)"
     modified_base   3188..3190
                     /created_by="User"
                     /label="R177H"
     modified_base   3434..3436
                     /created_by="User"
                     /modified_by="User"
                     /label="G95D"
     modified_base   3584..3586
                     /created_by="User"
                     /modified_by="User"
                     /label="P45T"
     promoter        complement(3807..3855)
                     /created_by="User"
                     /modified_by="User"
                     /label="aph (KmR) promoter"
     primer_bind     3811..3831
                     /Sequence="AGGGCTTCCCAACCTTACCAG"
                     /Hairpin_Tm="48.5"
                     /Tm="61.7"
                     /Self_Dimer_Tm="4.5"
                     /%GC="57.1"
                     /created_by="primer3"
                     /label="pZSeq2_Kan"
     primer_bind     3902..3921
                     /created_by="User"
                     /Sequence="GCGTATCACGAGGCCCTTTC"
                     /Hairpin_Tm="46.5"
                     /Tm="61.2"
                     /Self_Dimer_Tm="None"
                     /%GC="60.0"
                     /created_by="primer3"
                     /modified_by="User"
                     /label="pZ_seq_frw"
ORIGIN      
        1 cacagctaac accacgtcgt ccctatctgc tgccctaggt ctatgagtgg ttgctggata
       61 actttacggg catgcataag gctcgtatga tatattcagg gagaccacaa cggtttccct
      121 ctacaaataa ttttgtttaa cttttctaga tttaagaagg agatatacat atgtctgagc
      181 tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac aaccaccact
      241 tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc atgagaatca
      301 aggcggtcga gggcggccct ttacccttcg ccttcgacat cctggctacc agcttcatgt
      361 acggcagcaa aaccttcatc aaccacaccc agggcatccc cgacttcttt aagcagtcct
      421 tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc gtgctgaccg
      481 ctacccagga caccagctta caggacggct gcttgatcta caacgtcaag atcagagggg
      541 tgaacttccc atccaacggc cctgtgatgc agaagaaaac attaggctgg gaggcctcca
      601 ccgagaccct gtaccccgct gacggcggcc tggaaggcag agccgacatg gccctgaagt
      661 tagtgggcgg gggccacctg atctgcaact tgaagaccac atacagatcc aagaaacccg
      721 ctaagaactt gaagatgccc ggcgtctact atgtggacag aagactggaa agaatcaagg
      781 aggccgacaa agagacctac gtcgagcagc acgaggtggc tgtggccaga tactgcgacc
      841 tgcctagcaa actggggcac agataaatgt ccagacctgc aggcatgcaa gctctagagg
      901 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgaaat
      961 gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt ttcatctgtg
     1021 catatggaca gttttccctt tgatatctaa cggtgaacag ttgttctact tttgtttgtt
     1081 agtcttgatg cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt
     1141 agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt
     1201 gagatcatgc ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat
     1261 ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttacgtaggt aggaatctga
     1321 tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt
     1381 tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc
     1441 tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt
     1501 caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa
     1561 cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc
     1621 ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc
     1681 cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa
     1741 aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa
     1801 gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt
     1861 agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt attggttata
     1921 agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc
     1981 cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag
     2041 ttcatttgct ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta
     2101 atcactatac caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt
     2161 tgtgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc
     2221 ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat
     2281 ttatagaata aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa
     2341 ctcactactt tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc
     2401 tctacaaaac agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa
     2461 atcgctgaat attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg
     2521 acattcagtt cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc
     2581 gccttttatg gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct
     2641 tctcagggcg ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc
     2701 agttcctgcc ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc
     2761 agactggcta atgcacccag taaggcagcg gtatcatcaa caggcttacc cgtcttactg
     2821 tcaaccagac ccgccaggat tctcatgcat aaaaaacgcc cggcggcaac cgagcgttct
     2881 gaattaatta atcatcgcga ctgcagagat cccagagtcc ttattagaag aactcgtcaa
     2941 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     3001 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     3061 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     3121 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     3181 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     3241 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     3301 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     3361 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     3421 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     3481 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca
     3541 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     3601 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     3661 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     3721 gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg
     3781 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
     3841 cagctggcaa ttccgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3901 ggcgtatcac gaggcccttt cgtcttcac
//
