LOCUS       pEB2-2ndVal_mNeptune2.5        3968 bp    DNA     circular UNA 27-DEC-2017
DEFINITION  pEB2 plasmid with 2ndVal mNeptune2.5.
ACCESSION   urn.local...ez-87ycevh
VERSION     urn.local...ez-87ycevh
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..905
                     /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="2ndVal mNeptune2.5"
     motif           174..176
                     /created_by="User"
                     /modified_by="User"
                     /label="2nd Val"
     motif           213..215
                     /created_by="User"
                     /label="M11T"
     motif           264..266
                     /created_by="User"
                     /label="S28H"
     motif           276..278
                     /created_by="User"
                     /modified_by="User"
                     /label="R32G"
     motif           303..305
                     /created_by="User"
                     /modified_by="User"
                     /label="M41N"
     motif           306..308
                     /created_by="User"
                     /modified_by="User"
                     /label="K42R"
     motif           363..365
                     /created_by="User"
                     /modified_by="User"
                     /label="S61C"
     motif           417..419
                     /created_by="User"
                     /modified_by="User"
                     /label="L79F"
     motif           459..461
                     /created_by="User"
                     /modified_by="User"
                     /label="I93V"
     motif           492..494
                     /created_by="User"
                     /modified_by="User"
                     /label="A104V"
     motif           516..518
                     /created_by="User"
                     /modified_by="User"
                     /label="N112D"
     motif           525..527
                     /created_by="User"
                     /modified_by="User"
                     /label="I115L"
     motif           543..545
                     /created_by="User"
                     /modified_by="User"
                     /label="I121L"
     motif           546..548
                     /created_by="User"
                     /modified_by="User"
                     /label="N122R"
     motif           573..575
                     /created_by="User"
                     /modified_by="User"
                     /label="S131P"
     motif           609..611
                     /created_by="User"
                     /modified_by="User"
                     /label="N143S"
     motif           618..620
                     /created_by="User"
                     /modified_by="User"
                     /label="M146T"
     motif           645..647
                     /created_by="User"
                     /modified_by="User"
                     /label="R155E"
     motif           651..653
                     /created_by="User"
                     /modified_by="User"
                     /label="H157R"
     motif           654..656
                     /created_by="User"
                     /modified_by="User"
                     /label="S158C"
     motif           657..659
                     /created_by="User"
                     /modified_by="User"
                     /label="Q159D"
     motif           687..689
                     /created_by="User"
                     /modified_by="User"
                     /label="Y169H"
     motif           699..701
                     /created_by="User"
                     /modified_by="User"
                     /label="S173N"
     motif           702..704
                     /created_by="User"
                     /modified_by="User"
                     /label="F174L"
     motif           756..758
                     /created_by="User"
                     /modified_by="User"
                     /label="F192V"
     motif           759..761
                     /created_by="User"
                     /modified_by="User"
                     /label="H193Y"
     motif           771..773
                     /created_by="User"
                     /modified_by="User"
                     /label="H197R"
     motif           801..803
                     /created_by="User"
                     /modified_by="User"
                     /label="K207N"
     motif           828..830
                     /created_by="User"
                     /modified_by="User"
                     /label="M216V"
     motif           840..842
                     /created_by="User"
                     /modified_by="User"
                     /label="K220R"
     motif           873..875
                     /created_by="User"
                     /modified_by="User"
                     /label="R231K"
     source          <876..>902
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     motif           876..881
                     /created_by="User"
                     /label="LN spacer"
     motif           882..902
                     /created_by="User"
                     /label="avGFP -C terminus"
     terminator      937..995
                     /created_by="User"
                     /modified_by="User"
                     /label="rrnB T1 terminator"
     rep_origin      complement(996..2879)
                     /created_by="User"
                     /modified_by="User"
                     /label="Origin of Rep. from pSC101"
     repeat_region   complement(1019..1035)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     primer_bind     complement(1057..1080)
                     /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(1137..1151)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     CDS             complement(1162..2112)
                     /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(2114..2119)
                     /regulatory_class="ribosome_binding_site"
                     /note="Shine-Dalgarno sequence"
                     /label="Shine-Dalgarno sequence regulatory"
     repeat_region   complement(2114..2136)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2139..2160)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     regulatory      complement(2143..2148)
                     /regulatory_class="promoter"
                     /note="-10 region"
                     /label="-10 region regulatory"
     regulatory      complement(2165..2170)
                     /regulatory_class="promoter"
                     /note="-35 region"
                     /label="-35 region regulatory"
     repeat_region   complement(2760..2777)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2783..2806)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2817..2833)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     repeat_region   complement(2846..2859)
                     /note="inverted repeat"
                     /label="inverted repeat repeat region"
     terminator      complement(2885..2950)
                     /created_by="User"
                     /modified_by="User"
                     /label="t(o) terminator from pSS9"
     CDS             complement(2963..3757)
                     /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   3227..3229
                     /created_by="User"
                     /label="R177H"
     modified_base   3473..3475
                     /created_by="User"
                     /modified_by="User"
                     /label="G95D"
     modified_base   3623..3625
                     /created_by="User"
                     /modified_by="User"
                     /label="P45T"
     promoter        complement(3846..3894)
                     /created_by="User"
                     /modified_by="User"
                     /label="aph (KmR) promoter"
     primer_bind     3850..3870
                     /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     3941..3960
                     /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 atggtgagta
      181 aaggagaaga actgattaag gagaacatgc acaccaagct gtacatggag ggcaccgtga
      241 acaaccacca cttcaagtgc acacacgagg gcgaaggcaa gccctacgag ggcacccaga
      301 ccaacagaat caaggtggtc gagggcggcc ctttaccctt cgccttcgac atcctggcta
      361 cctgcttcat gtacggcagc aagaccttca tcaaccacac ccagggcatc cccgacttct
      421 ttaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac gaagacgggg
      481 gcgtgctgac cgttacccag gacaccagcc tgcaggacgg ctgcctgatc tacaacgtca
      541 agctcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactgggct
      601 gggaggccag caccgagacc ctgtaccccg ctgacggcgg cctggaaggc agatgcgaca
      661 tggccctgaa gttagtgggc gggggccacc tgcactgcaa cttgaagacc acatacagat
      721 ccaagaaacc cgctaagaac ttgaagatgc ccggcgtcta cttcgtggac aggagactgg
      781 aaagaatcaa ggaggccgac aatgagacct acgtcgagca gcacgaggtg gctgtggcca
      841 ggtactgcga cctgcctagc aaactggggc acaaacttaa tggcatggat gaactataca
      901 aataaatgtc cagacctgca ggcatgcaag ctctagaggc atcaaataaa acgaaaggct
      961 cagtcgaaag actgggcctt tcgttttatc tgttgaaatg caccaaaaac tcgtaaaagc
     1021 tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag ttttcccttt
     1081 gatatctaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc ttcactgata
     1141 gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt tctctagtgt
     1201 ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatgct tactttgcat
     1261 gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta aagcatcgtg
     1321 tagtgttttt cttagtccgt tacgtaggta ggaatctgat gtaatggttg ttggtatttt
     1381 gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca tttgtctatc
     1441 tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa ccaccaattt
     1501 catattgctg taagtgttta aatctttact tattggtttc aaaacccatt ggttaagcct
     1561 tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat caaggctaat
     1621 ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc actcataaat
     1681 cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt ttatgaattt
     1741 ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta atttttcgct
     1801 tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca aaggattcct
     1861 gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac cataagcatt
     1921 ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata ccgtccgttc
     1981 tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata aaattagctt
     2041 ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt tgaaaacaac
     2101 taattcagac atacatctca attggtctag gtgattttaa tcactatacc aattgagatg
     2161 ggctagtcaa tgataattac tagtcctttt cctttgagtt gtgggtatct gtaaattctg
     2221 ctagaccttt gctggaaaac ttgtaaattc tgctagaccc tctgtaaatt ccgctagacc
     2281 tttgtgtgtt ttttttgttt atattcaagt ggttataatt tatagaataa agaaagaata
     2341 aaaaaagata aaaagaatag atcccagccc tgtgtataac tcactacttt agtcagttcc
     2401 gcagtattac aaaaggatgt cgcaaacgct gtttgctcct ctacaaaaca gaccttaaaa
     2461 ccctaaaggc ttaagtagca ccctcgcaag ctcgggcaaa tcgctgaata ttccttttgt
     2521 ctccgaccat caggcacctg agtcgctgtc tttttcgtga cattcagttc gctgcgctca
     2581 cggctctggc agtgaatggg ggtaaatggc actacaggcg ccttttatgg attcatgcaa
     2641 ggaaactacc cataatacaa gaaaagcccg tcacgggctt ctcagggcgt tttatggcgg
     2701 gtctgctatg tggtgctatc tgactttttg ctgttcagca gttcctgccc tctgattttc
     2761 cagtctgacc acttcggatt atcccgtgac aggtcattca gactggctaa tgcacccagt
     2821 aaggcagcgg tatcatcaac aggcttaccc gtcttactgt caaccagacc cgccaggatt
     2881 ctcatgcata aaaaacgccc ggcggcaacc gagcgttctg aattaattaa tcatcgcgac
     2941 tgcagagatc ccagagtcct tattagaaga actcgtcaag aaggcgatag aaggcgatgc
     3001 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     3061 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     3121 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     3181 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc
     3241 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     3301 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     3361 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     3421 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     3481 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     3541 tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca
     3601 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     3661 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     3721 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     3781 cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
     3841 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccgacgtct
     3901 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     3961 gtcttcac
//
