LOCUS       pCAG-loxPSTOPloxP-Lyn-EGFP        6530 bp    DNA     circular
            
DEFINITION  pCAG-loxPSTOPloxP-Lyn-EGFP
ACCESSION   pCAG-loxPSTOPloxP-ZsGreen	6500
VERSION     pCAG-loxPSTOPloxP-ZsGreen	6500.0
KEYWORDS    .

COMMENT
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                        <color blue="102" green="153" red="0"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="153" green="204" red="102"/>
                    </fill>
                </style>
                <location x="224" y="240"/>
                <dimension height="362" width="362"/>
            </element>
            <element id="1636921882" name="EBV_rev_primer"
                target="EBV_rev_primer:2278 - 2297" text="@N" type="21" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="692" y="630"/>
            </element>
            <element id="1636921882" name="EBV_rev_primer"
                target="EBV_rev_primer:2278 - 2297" type="21" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="1636921882" name="EBV_rev_primer" shape="1"
                target="EBV_rev_primer:2278 - 2297" type="21" xsi:type="symbol">
                <style name="Shape 1" shape="1" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="102" green="153" red="0"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="153" green="204" red="102"/>
                    </fill>
                </style>
                <location x="174" y="190"/>
                <dimension height="462" width="462"/>
            </element>
            <element id="1604986086" name="M13_pUC_rev_primer"
                target="M13_pUC_rev_primer:6481 - 6503" text="@N"
                type="21" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="405" y="55"/>
            </element>
            <element id="1604986086" name="M13_pUC_rev_primer"
                target="M13_pUC_rev_primer:6481 - 6503" type="21" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="1604986086" name="M13_pUC_rev_primer" shape="1"
                target="M13_pUC_rev_primer:6481 - 6503" type="21" xsi:type="symbol">
                <style name="Shape 1" shape="1" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="102" green="153" red="0"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="153" green="204" red="102"/>
                    </fill>
                </style>
                <location x="224" y="240"/>
                <dimension height="362" width="362"/>
            </element>
            <element id="880987917" name="M13_pUC_fwd_primer"
                target="M13_pUC_fwd_primer:3782 - 3760" text="@N"
                type="21" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="193" y="647"/>
            </element>
            <element id="880987917" name="M13_pUC_fwd_primer"
                target="M13_pUC_fwd_primer:3782 - 3760" type="21" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="880987917" name="M13_pUC_fwd_primer" shape="1"
                target="M13_pUC_fwd_primer:3782 - 3760" type="21" xsi:type="symbol">
                <style name="Shape 1" shape="1" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="102" green="153" red="0"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="153" green="204" red="102"/>
                    </fill>
                </style>
                <location x="174" y="190"/>
                <dimension height="462" width="462"/>
            </element>
            <element id="863401706" name="EBV_rev_primer"
                target="EBV_rev_primer:2024 - 2043" text="@N" type="21" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="688" y="536"/>
            </element>
            <element id="863401706" name="EBV_rev_primer"
                target="EBV_rev_primer:2024 - 2043" type="21" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="863401706" name="EBV_rev_primer" shape="1"
                target="EBV_rev_primer:2024 - 2043" type="21" xsi:type="symbol">
                <style name="Shape 1" shape="1" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="102" green="153" red="0"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="153" green="204" red="102"/>
                    </fill>
                </style>
                <location x="174" y="190"/>
                <dimension height="462" width="462"/>
            </element>
            <element id="1971845723" name="BGH_rev_primer"
                target="BGH_rev_primer:3466 - 3449" text="@N" type="21" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="275" y="721"/>
            </element>
            <element id="1971845723" name="BGH_rev_primer"
                target="BGH_rev_primer:3466 - 3449" type="21" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="1971845723" name="BGH_rev_primer" shape="1"
                target="BGH_rev_primer:3466 - 3449" type="21" xsi:type="symbol">
                <style name="Shape 1" shape="1" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="102" green="153" red="0"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="153" green="204" red="102"/>
                    </fill>
                </style>
                <location x="224" y="240"/>
                <dimension height="362" width="362"/>
            </element>
            <element id="223450876" name="lacZ_a"
                target="lacZ_a:3760 - 3908" text="@N" type="21" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="218" y="682"/>
            </element>
            <element id="223450876" name="lacZ_a"
                target="lacZ_a:3760 - 3908" type="21" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="223450876" name="lacZ_a" shape="1"
                target="lacZ_a:3760 - 3908" type="21" xsi:type="symbol">
                <style name="Shape 1" shape="1" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="102" green="153" red="0"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="153" green="204" red="102"/>
                    </fill>
                </style>
                <location x="199" y="215"/>
                <dimension height="412" width="412"/>
            </element>
            <element id="1603035376" name="loxP"
                target="loxP:1766 - 1733" text="@N" type="21" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="659" y="449"/>
            </element>
            <element id="1603035376" name="loxP"
                target="loxP:1766 - 1733" type="21" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="1603035376" name="loxP" shape="1"
                target="loxP:1766 - 1733" type="21" xsi:type="symbol">
                <style name="Shape 1" shape="1" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="102" green="153" red="0"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="153" green="204" red="102"/>
                    </fill>
                </style>
                <location x="224" y="240"/>
                <dimension height="362" width="362"/>
            </element>
            <element id="472464874" name="Membrane targeting motif"
                target="Membrane targeting motif:2669 - 2698" text="@N"
                type="87" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="565" y="681"/>
            </element>
            <element id="472464874" name="Membrane targeting motif"
                target="Membrane targeting motif:2669 - 2698" type="87" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="472464874" name="Membrane targeting motif"
                shape="1" target="Membrane targeting motif:2669 - 2698"
                type="87" xsi:type="symbol">
                <style name="Shape 1" shape="1" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="102" green="153" red="0"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="153" green="204" red="102"/>
                    </fill>
                </style>
                <location x="199" y="215"/>
                <dimension height="412" width="412"/>
            </element>
            <element id="2075160759" name="bGH_PA_terminator"
                target="bGH_PA_terminator:3452 - 3679" text="@N"
                type="43" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="247" y="666"/>
            </element>
            <element id="2075160759" name="bGH_PA_terminator"
                target="bGH_PA_terminator:3452 - 3679" type="43" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="2075160759" name="bGH_PA_terminator" shape="0"
                target="bGH_PA_terminator:3452 - 3679" type="43" xsi:type="symbol">
                <style name="Short Signal" shape="0" xsi:type="basicStyle">
                    <line width="3">
                        <color blue="204" green="102" red="51"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="255" green="255" red="255"/>
                    </fill>
                </style>
                <location x="199" y="215"/>
                <dimension height="412" width="412"/>
            </element>
            <element id="1584135095" name="SV40_PA_terminator"
                target="SV40_PA_terminator:2444 - 2575" text="@N"
                type="43" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="575" y="612"/>
            </element>
            <element id="1584135095" name="SV40_PA_terminator"
                target="SV40_PA_terminator:2444 - 2575" type="43" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="1584135095" name="SV40_PA_terminator" shape="0"
                target="SV40_PA_terminator:2444 - 2575" type="43" xsi:type="symbol">
                <style name="Short Signal" shape="0" xsi:type="basicStyle">
                    <line width="3">
                        <color blue="204" green="102" red="51"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="255" green="255" red="255"/>
                    </fill>
                </style>
                <location x="199" y="215"/>
                <dimension height="412" width="412"/>
            </element>
            <element id="234562470" name="SV40_PA_terminator"
                target="SV40_PA_terminator:1936 - 2067" text="@N"
                type="43" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="645" y="509"/>
            </element>
            <element id="234562470" name="SV40_PA_terminator"
                target="SV40_PA_terminator:1936 - 2067" type="43" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="234562470" name="SV40_PA_terminator" shape="0"
                target="SV40_PA_terminator:1936 - 2067" type="43" xsi:type="symbol">
                <style name="Short Signal" shape="0" xsi:type="basicStyle">
                    <line width="3">
                        <color blue="204" green="102" red="51"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="255" green="255" red="255"/>
                    </fill>
                </style>
                <location x="199" y="215"/>
                <dimension height="412" width="412"/>
            </element>
            <element id="640025100" name="SV40_PA_terminator"
                target="SV40_PA_terminator:2190 - 2321" text="@N"
                type="43" xsi:type="label">
                <style name="Signal Label" xsi:type="basicStyle">
                    <label text="@N">
                        <font name="Arial" size="9" style="1"/>
                        <color blue="0" green="0" red="0"/>
                    </label>
                </style>
                <location x="658" y="592"/>
            </element>
            <element id="640025100" name="SV40_PA_terminator"
                target="SV40_PA_terminator:2190 - 2321" type="43" xsi:type="line">
                <style name="Signal Label" xsi:type="basicStyle">
                    <line width="1">
                        <color blue="0" green="0" red="0"/>
                        <pattern style="1"/>
                    </line>
                </style>
            </element>
            <element id="640025100" name="SV40_PA_terminator" shape="0"
                target="SV40_PA_terminator:2190 - 2321" type="43" xsi:type="symbol">
                <style name="Short Signal" shape="0" xsi:type="basicStyle">
                    <line width="3">
                        <color blue="204" green="102" red="51"/>
                        <pattern style="1"/>
                    </line>
                    <fill pattern="Solid">
                        <color blue="255" green="255" red="255"/>
                    </fill>
                </style>
                <location x="199" y="215"/>
                <dimension height="412" width="412"/>
            </element>
            <element name="Terminator" type="43" xsi:type="annotation">
                <label text="">
                    <style name="Signal Label" xsi:type="basicStyle">
                        <label text="@N">
                            <font name="Arial" size="9" style="1"/>
                            <color blue="0" green="0" red="0"/>
                        </label>
                        <line width="1">
                            <color blue="0" green="0" red="0"/>
                            <pattern style="1"/>
                        </line>
                    </style>
                </label>
                <connector>
                    <style name="Signal Label" xsi:type="basicStyle">
                        <label text="@N">
                            <font name="Arial" size="9" style="1"/>
                            <color blue="0" green="0" red="0"/>
                        </label>
                        <line width="1">
                            <color blue="0" green="0" red="0"/>
                            <pattern style="1"/>
                        </line>
                    </style>
                </connector>
                <symbol shape="0">
                    <style name="Short Signal" shape="13" xsi:type="basicStyle">
                        <line width="3">
                            <color blue="204" green="102" red="51"/>
                            <pattern style="1"/>
                        </line>
                        <fill pattern="Solid">
                            <color blue="255" green="255" red="255"/>
                        </fill>
                    </style>
                </symbol>
            </element>
        </elements>
    </setupInfo>
    <setupInfo complete="true" completeMinSize="150"
        completeNested="false" includeStopCodon="false"
        incomplete="false" incompleteMinSize="30" selected="false"
        startCodons="ATG GTG" stopCodons="TAA TGA TAG" type="ORFs"
        undefinedStart="false" undefinedStop="false" undefinedStopNested="false"/>
    <setupInfo allowBlunt="true" allowFivePrime="true"
        allowThreePrime="true"
        enzymeMap="AvaI|cycgrg,EcoRI|gaattc,SmaI|cccggg,PstI|ctgcag,ApaLI|gtgcac,NcoI|ccatgg,HindIII|aagctt,BamHI|ggatcc,XmaI|cccggg,ClaI|atcgat"
        enzymesCleavePoint="AvaI|2,EcoRI|2,SmaI|4,PstI|6,ApaLI|2,NcoI|2,HindIII|2,BamHI|2,XmaI|2,ClaI|3"
        enzymesComplCleavePoint="AvaI|0,EcoRI|0,SmaI|0,PstI|0,ApaLI|0,NcoI|0,HindIII|0,BamHI|0,XmaI|0,ClaI|0"
        enzymesPalindromic="AvaI|true,EcoRI|true,SmaI|true,PstI|true,ApaLI|true,NcoI|true,HindIII|true,BamHI|true,XmaI|true,ClaI|true"
        ignoreIn="-1,-1" ignoreLess="-1" ignoreMore="-1"
        ignoreOut="-1,-1" selected="true" startIndex="1" type="Restriction Map"/>
    <setupInfo featureList="" selected="true" type="Feature Map"/>
    <setupInfo basesPerBlock="10" blocksPerLine="10"
        prefer1LetterAACode="false" preferSingleStrand="false"
        selected="true" textBold="false" textColor="0,0,0"
        textFontName="Andale Mono" textFontSize="14" textItalic="false"
        textUnderline="false" trackSeparatorColor="128,128,128"
        trackSeparatorWidth="2" type="Sequence"
        xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:type="sequenceSetup">
        <textRangeSetupList/>
    </setupInfo>
    <setupInfo analysisSubsets="Analysis Results (MAIN)" selected="true" type="Auto Load Analysis"/>
</vntiDisplaySetup>
DISPLAY_SETUP_END|
COMMENT     Backbone is pCAG-loxPSTOPloxP-ZsGreen (addgene Plasmid #51269)
            
            pCAG-loxPSTOPloxP-ZsGreen was diggested with PstI/SacI
            
            Lyn-EGFP was amplified from PM-GFP (Plasmid #21213) using primers:
            
            PstI-loxP-STOP-pmGFP fw
            5’-AGGTCCCTCGACCTGCAGCTGCAGGCCACCATGGGATGTATTAAATC-3’

SacI-EGFP-Rv
            5’-TGATCAGCGAGCTCTTACTTGTACAGCTCGTCCATG-3’
            
            Product was LIC cloned into the vector.
            
            Resistance= Ampicilin
            VNTEXTCHREPL|Animal/Other Eukaryotic
            VNTREPLTYPE|Plasmid
FEATURES             Location/Qualifiers
     misc_feature    86..373
                     /label=CAG_enhancer
     CDS             complement(805..1536)
                     /label=ORF frame 3
                     /translation="MEKGTAARRRTKALPAHFLPAGAAPLRPRPLEGVRRRLPDFGSA
                     RFGTKEVPAPSRTITIKGNGCGSPRLDSRRRSGAAAPLPRALPGPRRPRPARHPHLPP
                     PPARHGEPRATPRTEPRTRSRAVLSNSGRGVQGGVTARPPRPHPLLTPPRTHPARSLC
                     SPRSPPAPRGTAPGRAPLAHTRSAQSPAPRPQRSQPPGSAGRTRRSPRTHTRTHPPSR
                     SPPHKGPSRSPLRLSRSHRKETSRH*"
     CDS             complement(846..1349)
                     /label=ORF frame 1
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
     promoter        1686..1704
                     /label=T7_promoter
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
     misc_feature    complement(1733..1766)
                     /label=loxP
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
     misc_feature    2024..2043
                     /label=EBV_rev_primer
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
     terminator      2190..2321
                     /label=SV40_PA_terminator
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
     misc_feature    2278..2297
                     /label=EBV_rev_primer
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
     terminator      2444..2575
                     /label=SV40_PA_terminator
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
     misc_feature    2532..2551
                     /label=EBV_rev_primer
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
     misc_feature    complement(2604..2637)
                     /label=loxP
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
     misc_feature    complement(3449..3466)
                     /label=BGH_rev_primer
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI
                     NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG
                     AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI
                     LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH
                     LTEHAIASGSALP*"
     terminator      3452..3679
                     /label=bGH_PA_terminator
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAI
                     NLCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDG
                     AVCICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGI
                     LKGDVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWH
                     LTEHAIASGSALP*"
     CDS             3653..4126
                     /label=ORF frame 2
                     /translation="MLGMRWALWLLRRKEPAGARDPLVLASRLERPPLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTR
                     SVTATLASALAPAPFAFFPSFLATFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*"
     promoter        complement(3751..3767)
                     /label=M13_forward20_primer
                     /translation="MLGMRWALWLLRRKEPAGARDPLVLASRLERPPLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTR
                     SVTATLASALAPAPFAFFPSFLATFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*"
     misc_feature    complement(3760..3782)
                     /label=M13_pUC_fwd_primer
                     /translation="MLGMRWALWLLRRKEPAGARDPLVLASRLERPPLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTR
                     SVTATLASALAPAPFAFFPSFLATFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*"
     misc_feature    3760..3908
                     /label=lacZ_a
                     /translation="MLGMRWALWLLRRKEPAGARDPLVLASRLERPPLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTR
                     SVTATLASALAPAPFAFFPSFLATFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*"
     rep_origin      3926..4232
                     /label=f1_origin
                     /translation="MLGMRWALWLLRRKEPAGARDPLVLASRLERPPLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTR
                     SVTATLASALAPAPFAFFPSFLATFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*"
     promoter        4425..4453
                     /label=AmpR_promoter
                     /translation="MLGMRWALWLLRRKEPAGARDPLVLASRLERPPLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTR
                     SVTATLASALAPAPFAFFPSFLATFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*"
     gene            4495..5355
                     /label=Ampicillin
                     /gene="Ampicillin"
                     /translation="MLGMRWALWLLRRKEPAGARDPLVLASRLERPPLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGALSAAGVVVTR
                     SVTATLASALAPAPFAFFPSFLATFAGFPRQALNRGLPLGFRFSALRHLDPKKLD*"
     CDS             4495..5355
                     /label=ORF frame 1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     rep_origin      5510..6129
                     /label=pBR322_origin
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        6438..6467
                     /label=lac_promoter
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    6481..6503
                     /label=M13_pUC_rev_primer
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        6502..6520
                     /label=M13_reverse_primer
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     terminator      1936..2067
                     /label=SV40_PA_terminator
                     /translation="MAAARRASTAAGAPGPPRPSPEPSPARGGPAPPGTPTCRHPPPG
                     TASPAPRPARSPAPEAGPCSATRGGGCRGGLQPDRRAHTPCSPPHAHTPHAAFVPLAA
                     PPHRGAPPPAALPSRTRGAHKAPRRARSAHSRRAARAARGAPHAHTHARTPRAAPPRT
                     KGPPGAL*"
                     /type="terminator"
                     /vntifkey="43"
     misc_structure  1936..2575
                     /label=STOP cassette
                     /type="misc_structure"
                     /vntifkey="88"
     eukaryotic      86..1536
                     /label=CAG promoter
                     /promoter_type="eukaryotic"
                     /type="eukaryotic"
                     /vntifkey="29"
     misc_signal     2669..2698
                     /label=Membrane targeting motif
                     /note="MGCIKSKRKD peptide from Lym kinase"
                     /type="misc_signal"
                     /vntifkey="87"
     misc_marker     2714..3433
                     /label=EGFP
                     /type="misc_marker"
                     /vntifkey="22"
     misc_feature    2669..3433
                     /label=Lyn-EGFP
                     /note="EGFP tagged with the membrane target domain of
                     Lyn-Kinase"
                     /type="misc_feature"
                     /vntifkey="21"
ORIGIN
        1 gattcgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
      541 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
      601 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
      661 ggcgggagtc gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga
      841 aagccttaaa gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tgcggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     1261 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctggcgga gccgaaatct gggaggcgcc gccgcacccc
     1441 ctctagcggg cgcgggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccatctcc agcctcgggg ctgccgcagg
     1561 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1621 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagatcctta
     1681 attaataata cgactcacta tagggggtcg acccgccacc aatcatttaa atataacttc
     1741 gtataatgta tgctatacga agttattcgc gatgaataaa tgaaagcttg cagatctgcg
     1801 actctagagg atctgcgact ctagaggatc ataatcagcc ataccacatt tgtagaggtt
     1861 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     1921 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1981 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2041 atcaatgtat cttatcatgt ctggatctgc gactctagag gatcataatc agccatacca
     2101 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     2161 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     2221 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2281 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgcgactct agaggatcat
     2341 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     2401 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     2461 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2521 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccccat
     2581 caagctgatc cggaaccctt aatataactt cgtataatgt atgctatacg aagttattag
     2641 gtccctcgac ctgcagctgc aggccaccat gggatgtatt aaatcaaaaa ggaaagacgg
     2701 ggcccggtac catatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2761 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2821 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2881 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2941 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3001 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3061 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3121 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     3181 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     3241 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3301 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     3361 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3421 gctgtacaag taagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3481 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3541 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3601 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3661 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc gagatccact
     3721 agttctagcc tcgaggctag agcggccgcc actggccgtc gttttacaac gtcgtgactg
     3781 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     3841 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     3901 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3961 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4021 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4081 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4141 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4201 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4261 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4321 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     4381 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4441 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4501 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4561 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4621 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4681 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4741 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4801 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4861 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4921 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4981 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5041 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5101 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5161 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5221 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5281 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5341 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5401 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5461 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5521 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5581 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5641 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     5701 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5761 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5821 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5881 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5941 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6001 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6061 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6121 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6181 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6241 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6301 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6361 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6421 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6481 agcggataac aatttcacac aggaaacagc tatgaccatg aggcgcgccg
//
