LOCUS       pEB2-mNeptune2          3965 bp    DNA     circular UNA 27-DEC-2017
DEFINITION  pEB2 plasmid with mNeptune2.
ACCESSION   urn.local...de-87ybfb1
VERSION     urn.local...de-87ybfb1
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..902
                     /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="mNeptune2"
     motif           171..188
                     /created_by="User"
                     /modified_by="User"
                     /label="avGFP_N_terminus"
     motif           273..275
                     /created_by="User"
                     /modified_by="User"
                     /label="R32G"
     motif           300..302
                     /created_by="User"
                     /modified_by="User"
                     /label="M41G"
     motif           303..305
                     /created_by="User"
                     /modified_by="User"
                     /label="K42R"
     motif           360..362
                     /created_by="User"
                     /modified_by="User"
                     /label="S61C"
     motif           414..416
                     /created_by="User"
                     /modified_by="User"
                     /label="L79F"
     motif           456..458
                     /created_by="User"
                     /modified_by="User"
                     /label="I93V"
     motif           489..491
                     /created_by="User"
                     /modified_by="User"
                     /label="A104V"
     motif           513..515
                     /created_by="User"
                     /modified_by="User"
                     /label="N112D"
     motif           522..524
                     /created_by="User"
                     /modified_by="User"
                     /label="I115L"
     motif           540..542
                     /created_by="User"
                     /modified_by="User"
                     /label="I121L"
     motif           543..545
                     /created_by="User"
                     /modified_by="User"
                     /label="N122R"
     motif           570..572
                     /created_by="User"
                     /modified_by="User"
                     /label="S131P"
     motif           606..608
                     /created_by="User"
                     /modified_by="User"
                     /label="N143S"
     motif           615..617
                     /created_by="User"
                     /modified_by="User"
                     /label="M146T"
     motif           642..644
                     /created_by="User"
                     /modified_by="User"
                     /label="R155E"
     motif           648..650
                     /created_by="User"
                     /modified_by="User"
                     /label="H157R"
     motif           651..653
                     /created_by="User"
                     /modified_by="User"
                     /label="S158C"
     motif           654..656
                     /created_by="User"
                     /modified_by="User"
                     /label="Q159D"
     motif           684..686
                     /created_by="User"
                     /modified_by="User"
                     /label="Y169H"
     motif           696..698
                     /created_by="User"
                     /modified_by="User"
                     /label="S173N"
     motif           699..701
                     /created_by="User"
                     /modified_by="User"
                     /label="F174L"
     motif           753..755
                     /created_by="User"
                     /modified_by="User"
                     /label="F192V"
     motif           756..758
                     /created_by="User"
                     /modified_by="User"
                     /label="H193Y"
     motif           768..770
                     /created_by="User"
                     /modified_by="User"
                     /label="H197R"
     motif           798..800
                     /created_by="User"
                     /modified_by="User"
                     /label="K207N"
     motif           825..827
                     /created_by="User"
                     /modified_by="User"
                     /label="M216V"
     motif           837..839
                     /created_by="User"
                     /modified_by="User"
                     /label="K220R"
     motif           870..872
                     /created_by="User"
                     /modified_by="User"
                     /label="R231K"
     motif           873..878
                     /created_by="User"
                     /label="LN spacer"
     motif           879..899
                     /created_by="User"
                     /label="avGFP -C terminus"
     terminator      934..992
                     /created_by="User"
                     /modified_by="User"
                     /label="rrnB T1 terminator"
     rep_origin      complement(993..2876)
                     /created_by="User"
                     /modified_by="User"
                     /label="Origin of Rep. from pSC101"
     repeat_region   complement(1016..1032)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     primer_bind     complement(1054..1077)
                     /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(1134..1148)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     CDS             complement(1159..2109)
                     /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(2111..2116)
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Dalgarno sequence"
                     /label="Shine-Dalgarno sequence regulatory"
     repeat_region   complement(2111..2133)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2136..2157)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     regulatory      complement(2140..2145)
                     /regulatory_class="promoter"
                     /note="-10 region"
                     /label="-10 region regulatory"
     regulatory      complement(2162..2167)
                     /regulatory_class="promoter"
                     /note="-35 region"
                     /label="-35 region regulatory"
     repeat_region   complement(2757..2774)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2780..2803)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2814..2830)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2843..2856)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     terminator      complement(2882..2947)
                     /created_by="User"
                     /modified_by="User"
                     /label="t(o) terminator from pSS9"
     CDS             complement(2960..3754)
                     /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   3224..3226
                     /created_by="User"
                     /label="R177H"
     modified_base   3470..3472
                     /created_by="User"
                     /modified_by="User"
                     /label="G95D"
     modified_base   3620..3622
                     /created_by="User"
                     /modified_by="User"
                     /label="P45T"
     promoter        complement(3843..3891)
                     /created_by="User"
                     /modified_by="User"
                     /label="aph (KmR) promoter"
     primer_bind     3847..3867
                     /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     3938..3957
                     /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 atgagtaaag
      181 gagaagaact gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
      241 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagaccg
      301 gcagaatcaa ggtggtcgag ggcggccctt tacccttcgc cttcgacatc ctggctacct
      361 gcttcatgta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttta
      421 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
      481 tgctgaccgt tacccaggac accagcctgc aggacggctg cctgatctac aacgtcaagc
      541 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctgggctggg
      601 aggccagcac cgagaccctg taccccgctg acggcggcct ggaaggcaga tgcgacatgg
      661 ccctgaagtt agtgggcggg ggccacctgc actgcaactt gaagaccaca tacagatcca
      721 agaaacccgc taagaacttg aagatgcccg gcgtctactt cgtggacagg agactggaaa
      781 gaatcaagga ggccgacaat gagacctacg tcgagcagca cgaggtggct gtggccaggt
      841 actgcgacct gcctagcaaa ctggggcaca aacttaatgg catggatgaa ctatacaaat
      901 aaatgtccag acctgcaggc atgcaagctc tagaggcatc aaataaaacg aaaggctcag
      961 tcgaaagact gggcctttcg ttttatctgt tgaaatgcac caaaaactcg taaaagctct
     1021 gatgtatcta tcttttttac accgttttca tctgtgcata tggacagttt tccctttgat
     1081 atctaacggt gaacagttgt tctacttttg tttgttagtc ttgatgcttc actgatagat
     1141 acaagagcca taagaacctc agatccttcc gtatttagcc agtatgttct ctagtgtggt
     1201 tcgttgtttt tgcgtgagcc atgagaacga accattgaga tcatgcttac tttgcatgtc
     1261 actcaaaaat tttgcctcaa aactggtgag ctgaattttt gcagttaaag catcgtgtag
     1321 tgtttttctt agtccgttac gtaggtagga atctgatgta atggttgttg gtattttgtc
     1381 accattcatt tttatctggt tgttctcaag ttcggttacg agatccattt gtctatctag
     1441 ttcaacttgg aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat
     1501 attgctgtaa gtgtttaaat ctttacttat tggtttcaaa acccattggt taagcctttt
     1561 aaactcatgg tagttatttt caagcattaa catgaactta aattcatcaa ggctaatctc
     1621 tatatttgcc ttgtgagttt tcttttgtgt tagttctttt aataaccact cataaatcct
     1681 catagagtat ttgttttcaa aagacttaac atgttccaga ttatatttta tgaatttttt
     1741 taactggaaa agataaggca atatctcttc actaaaaact aattctaatt tttcgcttga
     1801 gaacttggca tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag gattcctgat
     1861 ttccacagtt ctcgtcatca gctctctggt tgctttagct aatacaccat aagcattttc
     1921 cctactgatg ttcatcatct gagcgtattg gttataagtg aacgataccg tccgttcttt
     1981 ccttgtaggg ttttcaatcg tggggttgag tagtgccaca cagcataaaa ttagcttggt
     2041 ttcatgctcc gttaagtcat agcgactaat cgctagttca tttgctttga aaacaactaa
     2101 ttcagacata catctcaatt ggtctaggtg attttaatca ctataccaat tgagatgggc
     2161 tagtcaatga taattactag tccttttcct ttgagttgtg ggtatctgta aattctgcta
     2221 gacctttgct ggaaaacttg taaattctgc tagaccctct gtaaattccg ctagaccttt
     2281 gtgtgttttt tttgtttata ttcaagtggt tataatttat agaataaaga aagaataaaa
     2341 aaagataaaa agaatagatc ccagccctgt gtataactca ctactttagt cagttccgca
     2401 gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta caaaacagac cttaaaaccc
     2461 taaaggctta agtagcaccc tcgcaagctc gggcaaatcg ctgaatattc cttttgtctc
     2521 cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat tcagttcgct gcgctcacgg
     2581 ctctggcagt gaatgggggt aaatggcact acaggcgcct tttatggatt catgcaagga
     2641 aactacccat aatacaagaa aagcccgtca cgggcttctc agggcgtttt atggcgggtc
     2701 tgctatgtgg tgctatctga ctttttgctg ttcagcagtt cctgccctct gattttccag
     2761 tctgaccact tcggattatc ccgtgacagg tcattcagac tggctaatgc acccagtaag
     2821 gcagcggtat catcaacagg cttacccgtc ttactgtcaa ccagacccgc caggattctc
     2881 atgcataaaa aacgcccggc ggcaaccgag cgttctgaat taattaatca tcgcgactgc
     2941 agagatccca gagtccttat tagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     3001 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     3061 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     3121 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     3181 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg
     3241 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     3301 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     3361 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     3421 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     3481 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     3541 tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt
     3601 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     3661 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     3721 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     3781 gatcagatct tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta
     3841 ctttgcaggg cttcccaacc ttaccagagg gcgccccagc tggcaattcc gacgtctaag
     3901 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     3961 ttcac
//
