LOCUS       GA413_pEXSISERS_FLuc_loxP-PuroR        7602 bp    DNA     circular SYN 10-JAN-2022
DEFINITION  pBluescript II KS(+) vector DNA, phagemid excised from lambda
            ZAPII.
ACCESSION   X52327
KEYWORDS    artificial sequence; cloning vector; expression vector; vector.
SOURCE      synthetic construct.
  ORGANISM  synthetic construct
            other sequences; artificial sequences.
REFERENCE   1
  AUTHORS   Short,J.M., Fernandez,J.M., Sorge,J.A. and Huse,W.D.
  TITLE     Lambda ZAP: a bacteriophage lambda expression vector with in vivo
            excision properties
  JOURNAL   Nucleic Acids Res. 16 (15), 7583-7600 (1988)
REFERENCE   2
  AUTHORS   Alting-Mees,M.A. and Short,J.M.
  TITLE     pBluescript II: gene mapping vectors
  JOURNAL   Nucleic Acids Res. 17 (22), 9494 (1989)
REFERENCE   3  (bases 1 to 2961)
  AUTHORS   Thomas,E.A.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-FEB-1990) Thomas E.A., Stratagene Clonin Systems,
            11099 North Torney Pines Rd., La Jolla, CA 92037, USA
FEATURES             Location/Qualifiers
     misc_feature    join(complement(<5397..5457),complement(460..>657))
                     /vntifkey=21
                     /modified_by="User"
                     /label="lacZ'"
     Comment         634..657
                     /created_by="User"
                     /label="Gibson Overlap"
     Primer          644..682
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="N-NrdJ-1_AP6_1_fw"
     Comment         644..665
                     /created_by="User"
                     /label="Gibson Overlap"
     Comment         653..>657
                     /created_by="User"
                     /label="SacI"
     Comment         658..663
                     /created_by="User"
                     /label="BbsI"
     primer_bind     665..682
                     /Sequence="CTGTCTCGTGGGCAGCAG"
                     /Hairpin_Tm="None"
                     /Tm="60.4"
                     /Self_Dimer_Tm="None"
                     /%GC="66.7"
                     /created_by="primer3"
                     /modified_by="jeffe"
                     /label
     ORF             join(<666..2861,5110..>5388)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label
     ORF             join(<666..2853,5153..>5388)
                     /created_by="jeffe"
                     /label
     region          666..980
                     /created_by="User"
                     /modified_by="User"
                     /Transferred_From="Clipboard Contents 3 (back translated)"
                     /Transferred_Similarity="92.38%"
                     /label="N-NrdJ-1 Intein"
     Primer          complement(743..764)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="seqprimer\NrdJ-1_N_1_rv"
     Primer          875..896
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="seqprimer\NrdJ-1_N_1_fw"
     primer          960..1016
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="N-NrdJ-1_AP4_1_fw"
     primer          complement(960..981)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="N-NrdJ-1_1_rv"
     Comment         960..980
                     /created_by="jeffe"
                     /label="Gibson Overlap"
     Primer          complement(966..991)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="N-NrdJ-1_AP6_1_rv"
     primer_bind     993..1016
                     /Sequence="CCTGAGGATGAACTTGCTGCCAAC"
                     /Hairpin_Tm="None"
                     /Tm="64.2"
                     /Self_Dimer_Tm="None"
                     /%GC="54.2"
                     /created_by="primer3"
                     /modified_by="jeffe"
                     /label
     region          <993..1088
                     /gene="AP4 coiled coils"
                     /Transferred_From="pCAG_RnMVP_HLH-SnoopTag"
                     /Transferred_Similarity="90.91%"
                     /modified_by="User"
     region          1002..1085
                     /created_by="User"
                     /modified_by="User"
                     /Transferred_From="pCAG_RnMVP_HLH-SnoopTag"
                     /Transferred_Similarity="91.67%"
                     /label="7x gabcdef"
     misc_feature    complement(1070..1090)
                     /Sequence="CCGCCGTACTTAATGGCCTGG"
                     /Hairpin_Tm="55.1"
                     /Tm="63.4"
                     /Self_Dimer_Tm="3.3"
                     /%GC="61.9"
                     /created_by="primer3"
                     /modified_by="jeffe"
                     /label
                     /note="Geneious type: primer_bind_reverse"
     primer          complement(1070..1111)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="AP4_1_rv"
     Comment         1073..>1088
                     /created_by="jeffe"
                     /label="Gibson Overlap"
     Comment         1089..1094
                     /created_by="User"
                     /Transferred_From="pBluescript\II\KS+ (modified)
                     N-gp41-1-Int_AP6_OLLAS_NanoLuc_P5_C-gp41-1-Int empty"
                     /Transferred_Similarity="100.00%"
                     /label="BamHI"
     primer          1094..1149
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="FLuc_1_fw"
     Comment         1094..1111
                     /created_by="jeffe"
                     /label="Gibson Overlap"
     Comment         1110..1115
                     /created_by="User"
                     /Transferred_From="pBluescript\II\KS+ (modified)
                     N-gp41-1-Int_AP6_OLLAS_NanoLuc_P5_C-gp41-1-Int empty"
                     /Transferred_Similarity="100.00%"
                     /modified_by="jeffe"
                     /label="KpnI"
     misc_feature    1112
                     /Original_Bases="A"
                     /label="A"
                     /note="Geneious type: Editing History Replacement"
     Comment         1113..1118
                     /created_by="jeffe"
                     /label="was a AgeI site"
     misc_feature    1118
                     /Original_Bases="T"
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     primer_bind     1119..1149
                     /Sequence="ATGGAAGATGCCAAGAACATCAAGAAGGGCC"
                     /Hairpin_Tm="45.2"
                     /Tm="68.7"
                     /Self_Dimer_Tm="None"
                     /%GC="48.4"
                     /created_by="primer3"
                     /modified_by="jeffe"
                     /label
     region          <1119..2768
                     /created_by="jeffe"
                     /Transferred_From="pBS-KSII(+)_fLuc-x5mod_V5-tag_orthoSUMO
                     2"
                     /Transferred_Similarity="89.94%"
                     /modified_by="jeffe"
                     /label="PpyLuc-x5mod"
     primer          1119..1145
                     /created_by="jeffe"
                     /label="seqprimer\PpyLuc-2_fw"
     Mutation        1158..1160
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /Transferred_From="pBS-KSII(+)_fLuc-x5mod_V5-tag_orthoSUMO
                     2"
                     /Transferred_Similarity="66.67%"
                     /label="F14R"
     primer          complement(1204..1225)
                     /created_by="jeffe"
                     /label="seqprimer\PpyLuc-1_rv"
     Mutation        1221..1223
                     /created_by="jeffe"
                     /Transferred_From="pBS-KSII(+)_fLuc-x5mod_V5-tag_orthoSUMO
                     2"
                     /Transferred_Similarity="100.00%"
                     /label="L35Q"
     Mutation        1662..1664
                     /created_by="jeffe"
                     /Transferred_From="pBS-KSII(+)_fLuc-x5mod_V5-tag_orthoSUMO
                     2"
                     /Transferred_Similarity="100.00%"
                     /label="V182K"
     Mutation        1812..1814
                     /created_by="jeffe"
                     /Transferred_From="pBS-KSII(+)_fLuc-x5mod_V5-tag_orthoSUMO
                     2"
                     /Transferred_Similarity="100.00%"
                     /label="I232K"
     primer          2018..2039
                     /modified_by="jeffe"
                     /label="seqprimer\PpyLuc-1_fw"
     Mutation        2511..2513
                     /created_by="jeffe"
                     /Transferred_From="pBS-KSII(+)_fLuc-x5mod_V5-tag_orthoSUMO
                     2"
                     /Transferred_Similarity="33.33%"
                     /label="F465R"
     Mutation        2760..2768
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /Transferred_From="pBS-KSII(+)_fLuc-x5mod_V5-tag_orthoSUMO
                     2"
                     /Transferred_Similarity="77.78%"
                     /label="SKL>IAV (removal cryptic peroxisomal targeting
                     signal)"
     Comment         2772..2777
                     /created_by="jeffe"
                     /label="BamHI"
     region          2784..2807
                     /created_by="User"
                     /modified_by="User"
                     /X-Transferred_From="NLS-SSLS (back translated)"
                     /X-Transferred_Similarity="95.83%"
                     /Transferred_From="pBS-KSII(+)_fLuc-x5mod_V5-tag_orthoSUMO
                     2"
                     /Transferred_Similarity="91.67%"
                     /label="FLAG-tag"
     primer          2784..2808
                     /modified_by="jeffe"
                     /label="seqprimer\FLAG_1_fw"
     Comment         2809..2815
                     /created_by="jeffe"
                     /label="BstEII"
     region          complement(2820..2853)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="lox5171"
     Comment         2853..2858
                     /created_by="jeffe"
                     /label="BspEI"
     primer_bind     2859..2911
                     /created_by="jeffe"
                     /Sequence="CAGGTAAGTATCCTTTTTACAGCACAACTTAATGAGACAGATAGAAA
                     CTGGTC"
                     /modified_by="jeffe"
                     /label
     primer          2859..2911
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="bpA-PuroR-EF1a_for_fLuc-SUMO_1_fw"
     motif           2860..2861
                     /created_by="jeffe"
                     /label="Consensus SD"
     motif           2862..2867
                     /created_by="jeffe"
                     /label="Splice Donor"
     Comment         2862..2928
                     /created_by="jeffe"
                     /label="5'"
     misc_feature    complement(2902..2929)
                     /created_by="jeffe"
                     /Sequence="CCTACTCTGTTTCTACAAGACCAGTTTC"
                     /Hairpin_Tm="32.9"
                     /Tm="61.4"
                     /Self_Dimer_Tm="None"
                     /%GC="42.9"
                     /created_by="primer3"
                     /modified_by="jeffe"
                     /label
                     /note="Geneious type: primer_bind_reverse"
     primer          complement(2902..2952)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="Invert_EF1a_PuroR_bpA_1_rv"
     Primer          <2929..2943
                     /created_by="User"
                     /modified_by="jeffe"
                     /label="bGH-pA_BbsIdel_F3_1_rv"
     Comment         complement(2929..2934)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="BmtI"
     Comment         2929..2934
                     /created_by="jeffe"
                     /label="NheI"
     Comment         2933..2952
                     /created_by="jeffe"
                     /label="Gibson Overlap"
     polyA_signal    complement(2935..3206)
                     /X-Transferred_From="eEF1a/puro/FRT"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="eEF1_puro_FRT - optimized"
                     /Transferred_Similarity="100.00%"
                     /modified_by="jeffe"
                     /label="bGH-pA"
     misc_feature    complement(2936..2959)
                     /X-Transferred_From="eEF1a/puro/FRT"
                     /X-Transferred_Similarity="100.00%"
                     /Transferred_From="eEF1_puro_FRT - optimized"
                     /Transferred_Similarity="100.00%"
                     /label="primer B5`"
     Comment         complement(3000..3005)
                     /created_by="jeffe"
                     /label="BbsIdel"
     primer_bind     complement(3181..3206)
                     /Sequence="GCTCGCTGATCAGCCTCGACTGTGCC"
                     /Hairpin_Tm="51.3"
                     /Tm="71.1"
                     /Self_Dimer_Tm="35.8"
                     /%GC="65.4"
                     /created_by="primer3"
                     /label
     Primer          complement(3181..3206)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="bGH-pA_BbsIdel_1_fw"
     misc_feature    3187..3206
                     /Sequence="GTCGAGGCTGATCAGCGAGC"
                     /Hairpin_Tm="54.6"
                     /Tm="63.3"
                     /Self_Dimer_Tm="35.0"
                     /%GC="65.0"
                     /created_by="primer3"
                     /label
                     /note="Geneious type: primer_bind_reverse"
     Primer          3187..3206
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="EF1a_Puro_1_rv"
     Comment         3187..3206
                     /created_by="jeffe"
                     /label="Gibson Overlap"
     Comment         3207..3214
                     /created_by="jeffe"
                     /label="MreI"
     ORF             complement(3233..3832)
                     /Imported_from="<a
                     href=""http://wishart.biology.ualberta.ca/PlasMapper"">Pla
                     sMapper</a>"
                     /X-Transferred_From="eEF1a/puro/FRT"
                     /X-Transferred_Similarity="79.33%"
                     /Transferred_From="eEF1_puro_FRT - optimized"
                     /Transferred_Similarity="96.67%"
                     /modified_by="jeffe"
                     /label="puro ORF"
     Comment         3287..3292
                     /created_by="jeffe"
                     /label="BmgBI"
     Primer          3493..3511
                     /created_by="jeffe"
                     /label="seqprimer\PuroR_1_rv"
     Kozak           complement(3833..3838)
                     /created_by="User"
                     /modified_by="jeffe"
     Restriction     3836..3843
                     /created_by="User"
                     /label="AscI"
     Comment         complement(3836..3843)
                     /created_by="jeffe"
                     /label="AscI"
     mrna            complement(<3844..>3858)
                     /gene="EEF1A1"
                     /gene_synonym="CCS-3; CCS3; EE1A1; EEF-1; EEF1A; EF-Tu;
                     EF1A; GRAF-1EF; HNGC:16303; LENG7; PTI1; eEF1A-1"
                     /product="eukaryotic translation elongation factor 1 alpha
                     1"
                     /note="Derived by automated computational analysis using
                     gene prediction method: BestRefSeq."
                     /transcript_id="NM_001402.5"
                     /db_xref="GI:83367078; GeneID:1915; HGNC:HGNC:3189;
                     HPRD:00559; MIM:130590"
                     /transcription="CTTTTTCGCAACGGGTTTGCCGCCAGAACACAGGTGTCGTGA
                     AAACTACCCCTAAAAGCCAAAATGGGAAAGGAAAAGACTCATATCAACATTGTCGTCA
                     TTGGACACGTAGATTCGGGCAAGTCCACCACTACTGGCCATCTGATCTATAAATGCGG
                     TGGCATCGACAAAAGAACCATTGAAAAATTTGAGAAGGAGGCTGCTGAGATGGGAAAG
                     GGCTCCTTCAAGTATGCCTGGGTCTTGGATAAACTGAAAGCTGAGCGTGAACGTGGTA
                     TCACCATTGATATCTCCTTGTGGAAATTTGAGACCAGCAAGTACTATGTGACTATCAT
                     TGATGCCCCAGGACACAGAGACTTTATCAAAAACATGATTACAGGGACATCTCAGGCT
                     GACTGTGCTGTCCTGATTGTTGCTGCTGGTGTTGGTGAATTTGAAGCTGGTATCTCCA
                     AGAATGGGCAGACCCGAGAGCATGCCCTTCTGGCTTACACACTGGGTGTGAAACAACT
                     AATTGTCGGTGTTAACAAAATGGATTCCACTGAGCCACCCTACAGCCAGAAGAGATAT
                     GAGGAAATTGTTAAGGAAGTCAGCACTTACATTAAGAAAATTGGCTACAACCCCGACA
                     CAGTAGCATTTGTGCCAATTTCTGGTTGGAATGGTGACAACATGCTGGAGCCAAGTGC
                     TAACATGCCTTGGTTCAAGGGATGGAAAGTCACCCGTAAGGATGGCAATGCCAGTGGA
                     ACCACGCTGCTTGAGGCTCTGGACTGCATCCTACCACCAACTCGTCCAACTGACAAGC
                     CCTTGCGCCTGCCTCTCCAGGATGTCTACAAAATTGGTGGTATTGGTACTGTTCCTGT
                     TGGCCGAGTGGAGACTGGTGTTCTCAAACCCGGTATGGTGGTCACCTTTGCTCCAGTC
                     AACGTTACAACGGAAGTAAAATCTGTCGAAATGCACCATGAAGCTTTGAGTGAAGCTC
                     TTCCTGGGGACAATGTGGGCTTCAATGTCAAGAATGTGTCTGTCAAGGATGTTCGTCG
                     TGGCAACGTTGCTGGTGACAGCAAAAATGACCCACCAATGGAAGCAGCTGGCTTCACT
                     GCTCAGGTGATTATCCTGAACCATCCAGGCCAAATAAGCGCCGGCTATGCCCCTGTAT
                     TGGATTGCCACACGGCTCACATTGCATGCAAGTTTGCTGAGCTGAAGGAAAAGATTGA
                     TCGCCGTTCTGGTAAAAAGCTGGAAGATGGCCCTAAATTCTTGAAGTCTGGTGATGCT
                     GCCATTGTTGATATGGTTCCTGGCAAGCCCATGTGTGTTGAGAGCTTCTCAGACTATC
                     CACCTTTGGGTCGCTTTGCTGTTCGTGATATGAGACAGACAGTTGCGGTGGGTGTCAT
                     CAAAGCAGTGGACAAGAAGGCTGCTGGAGCTGGCAAGGTCACCAAGTCTGCCCAGAAA
                     GCTCAGAAGGCTAAATGAATATTATCCCTAATACCTGCCACCCCACTCTTAATCAGTG
                     GTGGAAGAACGGTCTCAGAACTGTTTGTTTCAATTGGCCATTTAAGTTTAGTAGTAAA
                     AGACTGGTTAATGATAACAATGCATCGTAAAACCTTCAGAAGGAAAGGAGAATGTTTT
                     GTGGACCACTTTGGTTTTCTTTTTTGCGTGTGGCAGTTTTAAGTTATTAGTTTTTAAA
                     ATCAGTACTTTTTAATGGAAACAACTTGACCAAAAATTTGTCACAGAATTTTGAGACC
                     CATTAAAAAAGTTAAATGAGAAACCTGTGTGTTCCTTTGGTCAACACCGAGACATTTA
                     GGTGAAAGACATCTAATTCTGGTTTTACGAATCTGGAAACTTCTTGAAAATGTAATTC
                     TTGAGTTAACACTTCTGGGTGGAGAATAGGGTTGTTTTCCCCCCACATAATTGGAAGG
                     GGAAGGAATATCATTTAAAGCTATGGGAGGGTTGCTTTGATTACAACACTGGAGAGAA
                     ATGCAGCATGTTGCTGATTGCCTGTCACTAAAACAGGCCAAAAACTGAGTCCTTGTGT
                     TGCATAGAAAGCTTCATGTTGCTAAACCAATGTTAAGTGAATCTTTGGAAACAAAATG
                     TTTCCAAATTACTGGGATGTGCATGTTGAAACGTGGGTTAAAATGACTGGGCAGTGAA
                     AGTTGACTATTTGCCATGACATAAGAAATAAGTGTAGTGGCTAGTGTACACCCTATGA
                     GTGGAAGGGTCCATTTTGAAGTCAGTGGAGTAAGCTTTATGCCAGTTTGATGGTTTCA
                     CAAGTTCTATTGAGTGCTATTCAGAATAGGAACAAGGTTCTAATAGAAAAAGATGGCA
                     ATTTGAAGTAGCTATAAAATTAGACTAATCTACATTGCTTTTCTCCTGCAGAGTCTAA
                     TACCTTTTATGCTTTGATAATTAGCAGTTTGTCTACTTGGTCACTAGGAATGAAACTA
                     CATGGTAATAGGCTTAACAGGTGTAATAGCCCACTTACTCCTGAATCTTTAAGCATTT
                     GTGCATTTGAAAAATGCTTTTCGCGATCTTCCTGCTGGGATTACAGGCATGAGCCACT
                     GTGCCTGACCTCCCATATGTAAAAGTGTCTAAAGGTTTTTTTTTGGTTATAAAAGGAA
                     AATTTTTGCTTAAGTTTGAAGGATAGGTAAAATTAAAGGACATGCTTTCTGTTTGTGT
                     GATGGTTTTTAAAAATTTTTTTTAAGATGGAGTTCTTGTTGCCCAGGCTAGAATGCAA
                     TGGCAAAATCTCACTGCAATCTCCTCCTCCTGGGTTCAAGCAATTCTCCTACTTCAGC
                     CTCCCAAGTAGCTGGGATTACAGGCATGTGCTAATTTGGTGTTTTTAATAGAGATGAG
                     GTTTTTCCATGTTGGTCAGGCTGGTCTCAAACTCCTGACCTTAGGTGATCGCCTCGGC
                     CTCCTAAAGTGCTGGAATTACAGGCATGAGCCACCATGCCTGGCCAGGACATGTGTTC
                     TTAAGGACATGCTAAGCAGGAGTTAAAGCAGCCCAAGAGATAAGGCCTCTTAAAGTGA
                     CTGGCAATGTGTATTGCTCAAGATTCAAAGGTACTTGAATTGGCCATAGACAAGTCTG
                     TAATGAAGTGTTATCGTTTTCCCTCATCTGAGTCTGAATTAGATAAAATGCCTTCCCA
                     TCAGCCAGTGCTCTGAGGTATCAAGTCTAAATTGAACTAGAGATTTTTGTCCTTAGTT
                     TCTTTGCTATCTAATGTTTACACAAGTAAATAGTCTAAGATTTGCTGGATGACAGAAA
                     AAACAGGTAAGGCCTTTAATAGATGGCCAATAGATGCCCTGATAATGAAAGTTGACAC
                     CTGTAAGATTTACCAGTAGAGAATTCTTGACATGCAAGGAAGCAAGATTTAACTGAAA
                     AATTGTTCCCACTGGAAGCAGGAATGAGTCAGTTTACTTGCATATACTGAGATTGAGA
                     TTAACTTCCTGTGAAACCCAGTGTCTTAGACAACTGTGGCTTGAGCACCACCTGCTGG
                     TATTCATTACAAACTTGCTCACTACAATAAATGAATTTTAAGCTTTAAAAAAAAAAAA
                     AAAAAA"
                     /Transferred_From="EEF1A1 - 1915"
                     /Transferred_Similarity="100.00%"
     promoter        complement(3859..5046)
                     /created_by="User"
                     /modified_by="jeffe"
                     /label="eEF1a"
     primer          4058..4083
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="seqprimer\EF1a_1_rv"
     primer          4540..4563
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="seqprimer\EF1a_2_rv"
     Comment         complement(4965..4970)
                     /created_by="jeffe"
                     /label="MfeI"
     primer          5028..5068
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="Invert_EF1a_PuroR_bpA_2_fw"
     Comment         5028..5045
                     /created_by="jeffe"
                     /label="Gibson Overlap"
     Comment         complement(5044..5049)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="BstBI"
     primer_bind     5050..5068
                     /Sequence="TCGCCTGCTTTTCTGCCTG"
                     /Hairpin_Tm="None"
                     /Tm="61.0"
                     /Self_Dimer_Tm="None"
                     /%GC="57.9"
                     /created_by="primer3"
                     /label
     Comment         5050..5109
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="3'"
     Mutation        5067
                     /created_by="jeffe"
                     /label="A>T"
     motif           5070..5076
                     /created_by="jeffe"
                     /label="YNYURAC"
     motif           5070..5076
                     /created_by="jeffe"
                     /label="YNYYRAY"
     motif           5072..5076
                     /created_by="jeffe"
                     /label="CURAY"
     Comment         5075
                     /created_by="jeffe"
                     /label="Branch site"
     motif           5076..5105
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="Y-rich"
     misc_feature    complement(5098..5118)
                     /Sequence="GCGGCCGCCCTGCAGAAAAAG"
                     /Hairpin_Tm="33.0"
                     /Tm="67.5"
                     /Self_Dimer_Tm="39.9"
                     /%GC="66.7"
                     /created_by="primer3"
                     /label
                     /note="Geneious type: primer_bind_reverse"
     primer          complement(5098..5193)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="SA_lox5171_P3_rv"
     motif           5110
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="Consensus SA"
     region          complement(5119..5152)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="lox5171"
     Comment         5159..5165
                     /created_by="jeffe"
                     /label="BstEII"
     primer_bind     5173..5193
                     /Sequence="GAGATCCAGCAGCTGGAAGAG"
                     /Hairpin_Tm="58.7"
                     /Tm="60.2"
                     /Self_Dimer_Tm="41.5"
                     /%GC="57.1"
                     /created_by="primer3"
                     /modified_by="jeffe"
                     /label
     region          <5173..>5256
                     /gene="P3 coiled coil"
                     /Transferred_From="pCAG_RnMVP_HLH-SnoopTag"
                     /Transferred_Similarity="86.87%"
                     /modified_by="User"
     region          5173..5256
                     /created_by="User"
                     /Transferred_From="pCAG_RnMVP_HLH-SnoopTag"
                     /Transferred_Similarity="91.67%"
                     /modified_by="User"
                     /label="7xgabcdef"
     primer          5173..5193
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="P3_1_fw"
     Comment         5173..5193
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="Gibson Overlap"
     misc_feature    complement(5238..5258)
                     /Sequence="CCGTACTTCAGAGCCTGGTTC"
                     /Hairpin_Tm="31.5"
                     /Tm="60.4"
                     /Self_Dimer_Tm="1.8"
                     /%GC="57.1"
                     /created_by="primer3"
                     /label
                     /note="Geneious type: primer_bind_reverse"
     primer          complement(5238..5295)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="P3_C-NrdJ-1_1_rv"
     Primer          5258..5291
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="C-NrdJ-1_P5_1_fw"
     region          5269..5388
                     /created_by="User"
                     /Transferred_From="Clipboard Contents 3 (back translated)"
                     /Transferred_Similarity="93.33%"
                     /label="C-NrdJ-1 Intein"
     primer_bind     5272..5295
                     /Sequence="GAAGCCAAGACCTACATCGGCAAG"
                     /Hairpin_Tm="42.1"
                     /Tm="63.7"
                     /Self_Dimer_Tm="None"
                     /%GC="54.2"
                     /created_by="primer3"
                     /label
     primer          5272..5295
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="C-NrdJ-1_1_fw"
     Comment         5272..5295
                     /created_by="jeffe"
                     /label="Gibson Overlap"
     Primer          5286..5304
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="seqprimer\NrdJ-1_C_1_fw"
     Primer          complement(5292..5314)
                     /created_by="jeffe"
                     /modified_by="jeffe"
                     /label="seqprimer\NrdJ-1_C_1_rv"
     misc_feature    complement(5367..5388)
                     /Sequence="GTTGTGCACCAGGATGTCGTTG"
                     /Hairpin_Tm="None"
                     /Tm="62.5"
                     /Self_Dimer_Tm="15.7"
                     /%GC="54.5"
                     /created_by="primer3"
                     /modified_by="jeffe"
                     /label
                     /note="Geneious type: primer_bind_reverse"
     Primer          complement(5367..5413)
                     /created_by="jeffe"
                     /label="C-NrdJ-1_P5_1_rv"
     Comment         5388
                     /created_by="User"
                     /label="Gibson Overlap"
     Comment         5389..5413
                     /created_by="User"
                     /label="Gibson Overlap"
     Comment         complement(5391..5396)
                     /created_by="User"
                     /label="BbsI"
     Comment         5397..5422
                     /created_by="User"
                     /label="Gibson Overlap"
     Comment         <5397..5401
                     /created_by="User"
                     /label="KpnI"
     rep_origin      5799..6466
                     /vntifkey=33
                     /label="pUC\ori"
     misc_marker     complement(6617..7474)
                     /vntifkey=22
                     /label="Amp(R)"
ORIGIN      
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctgaa
      661 gactctgtct cgtgggcagc agcgagatca tcaccagaaa ctacggcaag accaccatca
      721 aagaggtggt cgagatcttc gacaacgaca agaacatcca ggtgctggcc ttcaacaccc
      781 acaccgacaa catcgagtgg gcccctatca aggccgctca gctgacaaga cctaacgccg
      841 agctggtgga actggaaatc gacacactgc acggcgtgaa aaccatcaga tgcacccctg
      901 atcaccccgt gtacaccaag aacagaggct acgtgcgggc cgacgagctg acagatgatg
      961 acgaactggt ggtggctatc ggcggcggag gacctgagga tgaacttgct gccaacgagg
     1021 aagaactgca acagaacgaa cagaagctgg cccagattaa gcagaagctc caggccatta
     1081 agtacggcgg atccggcgga ggcggatctg gtaccggaat ggaagatgcc aagaacatca
     1141 agaagggccc tgctcctaga taccctctgg aagatggaac cgctggcgag cagctgcaca
     1201 aggccatgaa gagatacgct caggtgcccg gcacaatcgc cttcacagat gcccacatcg
     1261 aagtgaacat cacctacgcc gagtacttcg agatgagcgt gcggctggcc gaagctatga
     1321 agcgatacgg cctgaacacc aaccacagaa tcgtcgtgtg cagcgagaac agcctccagt
     1381 tcttcatgcc tgtgctgggc gctctgttca tcggagtggc tgtggctcct gccaacgaca
     1441 tctacaacga gcgcgagctg ctgaacagca tgaacatcag ccagcctacc gtggtgttcg
     1501 tgtccaagaa gggactgcaa aagatcctga acgtgcagaa gaagctgccc atcatccaga
     1561 aaatcatcat catggacagc aagaccgact accagggctt ccagagcatg tataccttcg
     1621 tgaccagcca tctgccacca ggcttcaacg agtacgactt caagcccgag agcttcgaca
     1681 gagacaagac aatcgccctg atcatgaaca gcagcggctc taccggactg cctaaaggcg
     1741 ttgccctgcc tcacagaaca gcttgcgtca gattcagcca cgccagagat cccatcttcg
     1801 gcaaccagat caagcctgac accgctatcc tgagcgtggt gccttttcac cacggcttcg
     1861 gcatgttcac cacactgggc tacctgatct gcggcttcag agtggtgctg atgtatcgct
     1921 ttgaggaaga actgttcctg cggagcctcc aggactacaa gatccagtct gctctgctgg
     1981 tgcctactct gttcagcttc tttgccaaga gcaccctgat cgataagtac gacctgagca
     2041 acctgcacga gatcgcctct ggcggagccc ctctgtctaa agaagtgggc gaagccgtcg
     2101 ccaagagatt tcatctgccc ggcatcagac aaggctacgg actgaccgag acaaccagcg
     2161 ccatcctgat cacacctgag ggcgacgata agcctggcgc tgtgggaaaa gtggtgccat
     2221 tcttcgaggc taaggtggtg gacctggaca ccggcaaaac actgggagtg aatcagaggg
     2281 gcgagctgtg tgtcagaggc cctatgatca tgagcggcta cgtgaacaac cccgaggcca
     2341 ccaacgctct gatcgacaag gatggctggc tgcacagcgg cgacattgcc tactgggacg
     2401 aagatgagca cttcttcatc gtggacagac tgaagtccct gatcaagtac aagggctacc
     2461 aggtggcccc tgccgagctg gaatctatcc tgctccagca tcctaacatc cgcgacgctg
     2521 gtgttgctgg cctgcctgac gatgatgctg gcgaacttcc tgctgccgtg gtggtgctgg
     2581 aacacggcaa gaccatgacc gagaaagaaa tcgtggacta cgtggcctct caagtgacca
     2641 ccgccaagaa actgagaggc ggcgtggtgt ttgtggacga ggtgccaaaa ggcctgaccg
     2701 gcaagctgga cgccagaaag atcagagaga tcctcatcaa ggccaagaaa ggcggcaaga
     2761 tcgctgtcgg aggatccggc ggagactaca aggacgacga tgacaaaggg tcacctggca
     2821 taacttcgta tagtacacat tatacgaagt tatccggaca ggtaagtatc ctttttacag
     2881 cacaacttaa tgagacagat agaaactggt cttgtagaaa cagagtaggc tagcccccag
     2941 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattc
     3001 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     3061 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     3121 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     3181 ggcacagtcg aggctgatca gcgagccgcc ggcgtctaga gaattgatcc cctcaggcgc
     3241 caggctttct ggtcatgcac caggttctag ggccctcagg cacttccacg tcggcggtca
     3301 cggtgaagcc cagtctctcg tagaagggca ggtttctggg ggcgcttgtt tccaggaagg
     3361 cgggcacgcc agccctttca gcagcttcca ccccaggcag caccacagca gatcccagtc
     3421 ccttgccctg gtggtcaggt gacacgccca cggtggccag aaaccaggca ggctcttttg
     3481 gtctgtgggg ggccagcagg ccttccatct gctgctgggc agccagtcta gagccgctca
     3541 gctcggccat tctaggtccg atctcggcga acacagcgcc ggcttccaca gactcagggg
     3601 ttgtccacac agccacagcg gcgccatcat cggccaccca cactttgccg atgtccaggc
     3661 ccactctggt cagaaacagt tcctgcagct cggtcactct ctcgatgtgc cggtcggggt
     3721 ccacggtgtg tcttgtggca gggtaatcgg cgaaggcagc ggccagtgtc cgcacagctc
     3781 ttggcacatc gtccctggtg gccagccgca ctgtgggctt gtactcggtc atggtggcgc
     3841 gccttttagg ggtagttttc acgacacctg aaatggaaga aaaaaacttt gaaccactgt
     3901 ctgaggcttg agaatgaacc aagatccaaa ctcaaaaagg gcaaattcca aggagaatta
     3961 catcaagtgc caagctggcc taacttcagt ctccacccac tcagtgtggg gaaactccat
     4021 cgcataaaac ccctcccccc aacctaaaga cgacgtactc caaaagctcg agaactaatc
     4081 gaggtgcctg gacggcgccc ggtactccgt ggagtcacat gaagcgacgg ctgaggacgg
     4141 aaaggccctt ttcctttgtg tgggtgactc acccgcccgc tctcccgagc gccgcgtcct
     4201 ccattttgag ctccctgcag cagggccggg aagcggccat ctttccgctc acgcaactgg
     4261 tgccgaccgg gccagccttg ccgcccaggg cggggcgata cacggcggcg cgaggccagg
     4321 caccagagca ggccggccag cttgagacta cccccgtccg attctcggtg gccgcgctcg
     4381 caggccccgc ctcgccgaac atgtgcgctg ggacgcacgg gccccgtcgc cgcccgcggc
     4441 cccaaaaacc gaaataccag tgtgcagatc ttggcccgca tttacaagac tatcttgcca
     4501 gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact tatcgaaagc
     4561 agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc gcccaggcca
     4621 ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg aactctccca
     4681 cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagccagg ggcgtggaag
     4741 taattcaagg cacgcaaggg ccataacccg taaagaggcc aggcccgcgg gaaccacaca
     4801 cggcacttac ctgtgttctg gcggcaaacc cgttgcgaaa aagaacgttc acggcgacta
     4861 ctgcacttat atacggttct cccccaccct cgggaaaaag gcggagccag tacacgacat
     4921 cactttccca gtttaccccg cgccaccttc tctaggcacc ggttcaattg ccgacccctc
     4981 cccccaactt ctcggggact gtgggcgatg tgcgctctgc ccactgacgg gcaccggagc
     5041 caattcgaat cgcctgcttt tctgcctggt actaacttct ctcccctctc ctcttttctt
     5101 tttctgcagg gcggccgcat aacttcgtat agtacacatt atacgaagtt atctggcggg
     5161 tcacccgagg atgagatcca gcagctggaa gaggaaatcg cccagctgga acagaagaat
     5221 gccgctctga aagagaagaa ccaggctctg aagtacggag gcggaggcat ggaagccaag
     5281 acctacatcg gcaagctgaa gtccagaaag atcgtgtcca acgaggacac ctacgacatc
     5341 cagaccagca cacacaactt tttcgccaac gacatcctgg tgcacaactc gtcttcgtac
     5401 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     5461 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5521 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5581 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5641 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5701 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5761 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5821 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5881 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5941 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6001 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6061 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6121 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6181 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6241 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     6301 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6361 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6421 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6481 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6541 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6601 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6661 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6721 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6781 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6841 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6901 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6961 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7021 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7081 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7141 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7201 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7261 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7321 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7381 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7441 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7501 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7561 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc ac
//
