LOCUS       pSCL391                 5996 bp    DNA     circular UNA 02-AUG-2022
DEFINITION  Gibson/Homology Cloning ligation product.
ACCESSION   urn.local...kc-erhsijt
VERSION     urn.local...kc-erhsijt
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    join(5583..5996,1..90)
                     /label="CEN/ARS"
     promoter        127..231
                     /label="AmpR promoter"
     CDS             232..300
                     /label="AmpR"
     CDS             301..1092
                     /label="AmpR"
     rep_origin      1263..1851
                     /snapgene_direction="RIGHT"
                     /label="ori"
     primer_bind     1866..1885
                     /created_by="slopez"
                     /Sequence="GGCCTTTTTACGGTTCCTGG"
                     /Tm_(binding_region)="59.1"
                     /Tm_(with_extension)="69.0"
                     /%GC_(binding_region)="55.0"
                     /%GC_(with_extension)="52.5"
                     /Hairpin_Tm_(with_extension)="42.9"
                     /Self_Dimer_Tm_(with_extension)="1.2"
                     /Extension="AGCTGCATGTGTCAGAGGTT"
                     /modified_by="slopez"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="294-451 - 453-454 - 455-456 - 452-293"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL453
                     (<a href=""urn:local:.:u0-d55aa9h"">294-451 - 453-454 -
                     455-456 - 452-293</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL453 (<a href=""urn:local:.:6j-d558buk"">294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL453 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL453 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL453 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL453 (<a href=""urn:local:.:9y-d5593p5"">453-454</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL453 (<a href=""urn:local:.:v-d4vdlzu"">pSCL2</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL453 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL453 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL453 (<a href=""urn:local:.:km-d55a14i"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /label="SCL453"
     promoter        1907..2267
                     /label="pGal7"
     primer_bind     2191..2212
                     /%GC="40.9"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="57.6"
                     /created_by="primer3"
                     /Sequence="AGGTCGGAAATATTTATGGGCA"
                     /Mismatches=0
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="pSCL27"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL251
                     (<a href=""urn:local:.:i-cppv4as"">pSCL27</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:31h-d5h53rm"">pSCL73</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:q-cppvdfc"">pSCL27
                     (reversed)</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:10p-d55cf47"">pSCL70</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:10p-d55cf47"">pSCL70</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:vc-cylmts8"">pSCL27</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:24-ckph90c"">pSCL27</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:2o-ckphag8"">pSCL28</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:3y-d1qf8q0"">pSCL27</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:w-cppve81"">pSCL27
                     (reversed)</a>)"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pSCL77"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL251
                     (<a href=""urn:local:.:42w-d5qydkl"">pSCL77</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:4dm-d5utwmk"">pSCL80</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:479-d5r569u"">pSCL27</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:3iy-d5l0qh4"">pSCL74</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:49o-d5r8z74"">pSCL79</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a
                     href=""urn:local:.:2ct-d9qutxn"">pSCL92--re-design?</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:31h-d5h53rm"">pSCL73</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:3z6-d5qxtf9"">pSCL78</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:a1-ekqc2qq"">pSCL315</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL251 (<a href=""urn:local:.:q6-dc0im63"">pSCL101</a>)"
                     /label="SCL251"
     primer_bind     complement(2241..2267)
                     /%GC="25.9"
                     /Hairpin_Tm="33.4"
                     /Self_Dimer_Tm="17.1"
                     /Tm="57.1"
                     /created_by="primer3"
                     /Sequence="TTTTGAGGGAATATTCAACTGTTTTTT"
                     /Mismatches=0
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="922-923 (wt)"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL167
                     (<a href=""urn:local:.:ia-dv5iaof"">922-923 (wt)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL167 (<a href=""urn:local:.:7d-duuvir9"">pSCL27</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL167 (<a href=""urn:local:.:gc-dvb0jpa"">pSCL204</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL167 (<a href=""urn:local:.:24-ckph90c"">pSCL27</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL167 (<a href=""urn:local:.:30j-c3qkjq8"">pZS165</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL167 (<a href=""urn:local:.:10l-ckanc0u"">167-168</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL167 (<a href=""urn:local:.:3y-d1qf8q0"">pSCL27</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL167 (<a href=""urn:local:.:3we-d5qx0ep"">167-168</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL167 (<a href=""urn:local:.:3z6-d5qxtf9"">pSCL78</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL167 (<a href=""urn:local:.:12m-cc0z43x"">150-151</a>)"
                     /label="SCL167"
     primer_bind     complement(2242..2267)
                     /created_by="slopez"
                     /Sequence="TTTTGAGGGAATATTCAACTGTTTTT"
                     /Tm_(binding_region)="56.5"
                     /Tm_(with_extension)="66.4"
                     /%GC_(binding_region)="26.9"
                     /%GC_(with_extension)="37.0"
                     /Hairpin_Tm_(with_extension)="65.2"
                     /Self_Dimer_Tm_(with_extension)="24.6"
                     /Extension="TTTCTTAGCTGCCTATACGGCAGTGAAC"
                     /modified_by="slopez"
                     /label="oSCL328"
     primer_bind     complement(2242..2267)
                     /created_by="slopez"
                     /Sequence="TTTTGAGGGAATATTCAACTGTTTTT"
                     /Tm_(binding_region)="56.5"
                     /Tm_(with_extension)="64.5"
                     /%GC_(binding_region)="26.9"
                     /%GC_(with_extension)="39.5"
                     /Hairpin_Tm_(with_extension)="65.2"
                     /Self_Dimer_Tm_(with_extension)="24.6"
                     /Extension="GCTGCCTATACGGCAGT"
                     /modified_by="slopez"
                     /label="oSCL332"
     misc_feature    2268..2314
                     /label="HH-Ribozyme"
     primer_bind     complement(2291..2314)
                     /Sequence="GACGAGCTAGCTCGTTTCGTCCTC"
                     /Tm="65.0"
                     /%GC="58.3"
                     /Hairpin_Tm="66.2"
                     /Self_Dimer_Tm="51.7"
                     /created_by="slopez"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pSCL73"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL471
                     (<a href=""urn:local:.:31h-d5h53rm"">pSCL73</a>)"
                     /label="SCL471"
     primer_bind     complement(2295..2314)
                     /created_by="slopez"
                     /X-Transferred_From="449-450"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL449
                     (<a href=""urn:local:.:ez-d4vznr7"">449-450</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL449 (<a
                     href=""urn:local:.:f9-d4vzplw"">eco2-editing-retron</a>)"
                     /Sequence="GACGAGCTAGCTCGTTTCGT"
                     /Tm="60.2"
                     /%GC="55.0"
                     /Hairpin_Tm="66.2"
                     /Self_Dimer_Tm="51.7"
                     /modified_by="slopez"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="294-451 - 453-454 - 455-456 - 452-293"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL454
                     (<a href=""urn:local:.:u0-d55aa9h"">294-451 - 453-454 -
                     455-456 - 452-293</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL454 (<a href=""urn:local:.:6j-d558buk"">294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL454 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL454 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL454 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL454 (<a href=""urn:local:.:9y-d5593p5"">453-454</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL454 (<a href=""urn:local:.:v-d4vdlzu"">pSCL2</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL454 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL454 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL454 (<a href=""urn:local:.:km-d55a14i"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /label="SCL454"
     primer_bind     complement(2296..2315)
                     /Sequence="TGACGAGCTAGCTCGTTTCG"
                     /Tm="60.2"
                     /%GC="55.0"
                     /Hairpin_Tm="66.2"
                     /Self_Dimer_Tm="51.7"
                     /created_by="slopez"
                     /label="SCL257"
     misc_feature    2315
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /Transferred_From="CRISPEY_retron-guide"
                     /Transferred_Similarity="100.00%"
                     /label="msr"
     primer_bind     2315..2338
                     /created_by="slopez"
                     /Sequence="ATGATAAGATTCCGTATGCGCACC"
                     /Tm_(binding_region)="61.4"
                     /Tm_(with_extension)="68.8"
                     /%GC_(binding_region)="45.8"
                     /%GC_(with_extension)="46.2"
                     /Hairpin_Tm_(with_extension)="71.5"
                     /Self_Dimer_Tm_(with_extension)="27.8"
                     /Extension="GTTCACTGCCGTATAGGCAGCTAAGAAA"
                     /modified_by="slopez"
                     /label="oSCL330"
     primer_bind     2315..2338
                     /created_by="slopez"
                     /Sequence="ATGATAAGATTCCGTATGCGCACC"
                     /Tm_(binding_region)="61.4"
                     /Tm_(with_extension)="68.8"
                     /%GC_(binding_region)="45.8"
                     /%GC_(with_extension)="47.9"
                     /Hairpin_Tm_(with_extension)="71.5"
                     /Self_Dimer_Tm_(with_extension)="27.8"
                     /Extension="CTCAAAAACTGCCGTATAGGCAGC"
                     /modified_by="slopez"
                     /label="oSCL334"
     primer_bind     2315..2338
                     /created_by="slopez"
                     /Sequence="ATGATAAGATTCCGTATGCGCACC"
                     /Tm_(binding_region)="61.4"
                     /Tm_(with_extension)="77.3"
                     /%GC_(binding_region)="45.8"
                     /%GC_(with_extension)="37.5"
                     /Extension="CATGATAAAAAAAAACAGTTGAATATTCCCTCAAAAGTTCACTGCC
                     GTATAGGCAGCTAAGAAA"
                     /modified_by="slopez"
                     /label="oSCL338"
     primer_bind     2315..2338
                     /created_by="slopez"
                     /Sequence="ATGATAAGATTCCGTATGCGCACC"
                     /Tm_(binding_region)="61.4"
                     /Tm_(with_extension)="76.8"
                     /%GC_(binding_region)="45.8"
                     /%GC_(with_extension)="39.0"
                     /Extension="CATGATAAAAAAAAACAGTTGAATATTCCCTCAAAAACTGCCGTAT
                     AGGCAGC"
                     /modified_by="slopez"
                     /label="oSCL340"
     misc_feature    2316..2330
                     /Color="-1"
                     /label="crispey-ext"
     misc_feature    <2331..2411
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /Transferred_From="CRISPEY_retron-guide"
                     /Transferred_Similarity="100.00%"
                     /label="msr"
     misc_feature    2331..2342
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /Transferred_From="CRISPEY_retron-guide"
                     /Transferred_Similarity="100.00%"
                     /label="inverted repeat"
     primer_bind     complement(2397..2423)
                     /Sequence="AGGAAAACAGACAGTAACTCAGATTCA"
                     /Tm="60.4"
                     /%GC="37.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /created_by="slopez"
                     /label="SCL499"
     misc_RNA        2401..2544
                     /created_by="slopez"
                     /modified_by="slopez"
                     /label="msd"
     primer_bind     2401..2426
                     /Sequence="TCTGAGTTACTGTCTGTTTTCCTTGG"
                     /Tm="60.9"
                     /%GC="42.3"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /created_by="slopez"
                     /label="oSCL317"
     CDS             2424..2523
                     /created_by="slopez"
                     /modified_by="slopez"
                     /label="Ade2 donor"
     primer_bind     complement(2448..2473)
                     /Sequence="GGCAATTTCGTTAATAAGCAATTCCC"
                     /Tm="60.0"
                     /%GC="38.5"
                     /Hairpin_Tm="32.2"
                     /Self_Dimer_Tm="None"
                     /created_by="slopez"
                     /label="SCL984"
     primer_bind     2477..2496
                     /Sequence="AGGCCTCACAACTCTGGACA"
                     /Tm="60.8"
                     /%GC="55.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /created_by="slopez"
                     /label="SCL983"
     misc_feature    2524..2557
                     /ApEinfo_fwdcolor="#ffef86"
                     /ApEinfo_revcolor="#ffef86"
                     /Transferred_From="CRISPEY_retron-guide"
                     /Transferred_Similarity="100.00%"
                     /label="constant synthesis"
     primer_bind     2524..2545
                     /Sequence="AGGAAACCCGTTTCTTCTGACG"
                     /Tm="60.8"
                     /%GC="50.0"
                     /Hairpin_Tm="46.0"
                     /Self_Dimer_Tm="17.4"
                     /created_by="slopez"
                     /label="SCL541"
     primer_bind     complement(2538..2557)
                     /created_by="slopez"
                     /Sequence="TGCGCACCCTTACGTCAGAA"
                     /Tm_(binding_region)="62.1"
                     /Tm_(with_extension)="73.6"
                     /%GC_(binding_region)="55.0"
                     /%GC_(with_extension)="54.5"
                     /Hairpin_Tm_(with_extension)="34.5"
                     /Self_Dimer_Tm_(with_extension)="26.1"
                     /Extension="CTGGGACCATGCCGGCCATCTGATAAGATTCCGTA"
                     /modified_by="slopez"
                     /label="SCL487"
     CDS             2544
                     /created_by="slopez"
                     /label="RT start"
     misc_feature    complement(2546..2557)
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /Transferred_From="CRISPEY_retron-guide"
                     /Transferred_Similarity="100.00%"
                     /label="inverted repeat"
     primer_bind     complement(2550..2572)
                     /created_by="slopez"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="294-451 - 453-454 - 455-456 - 452-293"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL451
                     (<a href=""urn:local:.:u0-d55aa9h"">294-451 - 453-454 -
                     455-456 - 452-293</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL451 (<a href=""urn:local:.:6j-d558buk"">294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL451 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL451 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL451 (<a href=""urn:local:.:6j-d4vs398"">eco1 editing
                     retron</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL451 (<a href=""urn:local:.:73-d4vs9be"">eco1
                     crispey</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL451 (<a href=""urn:local:.:v-d4vdlzu"">pSCL2</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL451 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL451 (<a href=""urn:local:.:km-d55a14i"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /Sequence="TGATAAGATTCCGTATGCGCACC"
                     /Tm_(binding_region)="61.1"
                     /Tm_(with_extension)="71.6"
                     /%GC_(binding_region)="47.8"
                     /%GC_(with_extension)="58.1"
                     /Hairpin_Tm_(with_extension)="34.5"
                     /Self_Dimer_Tm_(with_extension)="23.6"
                     /Extension="CTGGGACCATGCCGGCCATC"
                     /modified_by="slopez"
                     /label="SCL451"
     misc_feature    complement(2558..2572)
                     /Color="-1"
                     /label="long primer"
     misc_difference 2573..2590
                     /created_by="Seth"
                     /label="gRNA (18 bases)"
     primer_bind     2573..2597
                     /created_by="slopez"
                     /X-Transferred_From="449-450"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL450
                     (<a href=""urn:local:.:ez-d4vznr7"">449-450</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL450 (<a
                     href=""urn:local:.:f9-d4vzplw"">eco2-editing-retron</a>)"
                     /Sequence="ATTAACGAAATTGCCCCAGTTTCAG"
                     /Tm_(binding_region)="60.6"
                     /Tm_(with_extension)="68.1"
                     /%GC_(binding_region)="40.0"
                     /%GC_(with_extension)="46.7"
                     /Hairpin_Tm_(with_extension)="60.8"
                     /Self_Dimer_Tm_(with_extension)="46.5"
                     /Extension="ACGAAACGAGCTAGCTCGTC"
                     /modified_by="slopez"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="452-294 - 453-454 - 455-456 - 293-451"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL455
                     (<a href=""urn:local:.:fm-d559pe1"">452-294 - 453-454 -
                     455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL455 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL455 (<a href=""urn:local:.:d8-d559d8i"">455-456</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL455 (<a href=""urn:local:.:v-d4vdlzu"">pSCL2</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL455 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL455 (<a href=""urn:local:.:km-d55a14i"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /label="SCL455"
     primer_bind     2573..2597
                     /created_by="slopez"
                     /Sequence="ATTAACGAAATTGCCCCAGTTTCAG"
                     /Tm="60.6"
                     /%GC="40.0"
                     /Hairpin_Tm="39.1"
                     /Self_Dimer_Tm="11.8"
                     /modified_by="slopez"
                     /label="SCL470"
     misc_feature    2591..2681
                     /ApEinfo_fwdcolor="#7ea7d8"
                     /ApEinfo_revcolor="#7ea7d8"
                     /label="modified SpCas9 scaffold"
     primer_bind     complement(2660..2681)
                     /created_by="slopez"
                     /Sequence="AAAAAGCACCGACTCGGTGCCA"
                     /Tm_(binding_region)="65.5"
                     /Tm_(with_extension)="71.6"
                     /%GC_(binding_region)="54.5"
                     /%GC_(with_extension)="50.0"
                     /Hairpin_Tm_(with_extension)="66.9"
                     /Self_Dimer_Tm_(with_extension)="43.3"
                     /Extension="TTTCTTAGCTGCCTATACGGCAGTGAAC"
                     /modified_by="slopez"
                     /label="oSCL314"
     primer_bind     complement(2662..2681)
                     /created_by="slopez"
                     /Sequence="AAAAAGCACCGACTCGGTGC"
                     /Tm_(binding_region)="61.8"
                     /Tm_(with_extension)="70.5"
                     /%GC_(binding_region)="55.0"
                     /%GC_(with_extension)="50.0"
                     /Hairpin_Tm_(with_extension)="67.0"
                     /Self_Dimer_Tm_(with_extension)="41.9"
                     /Extension="TTTCTTAGCTGCCTATACGGCAGTGAAC"
                     /modified_by="slopez"
                     /label="oSCL331"
     primer_bind     complement(2662..2681)
                     /Sequence="AAAAAGCACCGACTCGGTGC"
                     /Tm_(binding_region)="61.8"
                     /Tm_(with_extension)="69.6"
                     /%GC_(binding_region)="55.0"
                     /%GC_(with_extension)="51.1"
                     /Hairpin_Tm_(with_extension)="64.7"
                     /Self_Dimer_Tm_(with_extension)="41.9"
                     /Extension="CTTTCTTTGCTGCCTATACGGCAGT"
                     /created_by="slopez"
                     /label="oSCL335"
     primer_bind     complement(2662..2681)
                     /created_by="slopez"
                     /Sequence="AAAAAGCACCGACTCGGTGC"
                     /Tm_(binding_region)="61.8"
                     /Tm_(with_extension)="77.1"
                     /%GC_(binding_region)="55.0"
                     /%GC_(with_extension)="37.6"
                     /Extension="GAAAAAATATGATATGAATGAATATTCCACTTTCTTTTTTCTTAGC
                     TGCCTATACGGCAGTGAAC"
                     /modified_by="slopez"
                     /label="oSCL339"
     primer_bind     complement(2662..2681)
                     /created_by="slopez"
                     /Sequence="AAAAAGCACCGACTCGGTGC"
                     /Tm_(binding_region)="61.8"
                     /Tm_(with_extension)="76.6"
                     /%GC_(binding_region)="55.0"
                     /%GC_(with_extension)="39.2"
                     /Extension="GAAAAAATATGATATGAATGAATATTCCACTTTCTTTGCTGCCTAT
                     ACGGCAGT"
                     /modified_by="slopez"
                     /label="oSCL341"
     motif           2682..2709
                     /created_by="slopez"
                     /modified_by="slopez"
                     /X-Transferred_From="mKate2_EX1-[28-gRNA1-28]Hsv1-mKate2_E
                     X2-pA Copy"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; Csy4
                     site (SCL annotated) (<a
                     href=""urn:local:.:1rl-dk0jwbl"">mKate2_EX1-[28-gRNA1-28]H
                     sv1-mKate2_EX2-pA Copy</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Csy4 site (SCL annotated) (<a
                     href=""urn:local:.:1rl-dk0jwbl"">mKate2_EX1-[28-gRNA1-28]H
                     sv1-mKate2_EX2-pA Copy</a>)"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pRC04 _array"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; Csy4
                     site (SCL annotated) (<a
                     href=""urn:local:.:bg-erc3gj7"">pRC04 _array</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Csy4
                     site (SCL annotated) (<a
                     href=""urn:local:.:bg-erc3gj7"">pRC04 _array</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Csy4
                     site (SCL annotated) (<a
                     href=""urn:local:.:bg-erc3gj7"">pRC04 _array</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; Csy4
                     site (SCL annotated) (<a
                     href=""urn:local:.:bg-erc3gj7"">pRC04 _array</a>)"
                     /label="Csy4 site (SCL annotated)"
     primer_bind     2710..2737
                     /Sequence="TGATAAGATTCCGTATGCGCACCCTTAG"
                     /Tm_(binding_region)="64.7"
                     /Tm_(with_extension)="69.4"
                     /%GC_(binding_region)="46.4"
                     /%GC_(with_extension)="46.4"
                     /Hairpin_Tm_(with_extension)="71.5"
                     /Self_Dimer_Tm_(with_extension)="27.8"
                     /Extension="GTTCACTGCCGTATAGGCAGCTAAGAAA"
                     /created_by="slopez"
                     /label="oSCL315"
     primer_bind     2710..2736
                     /Sequence="TGATAAGATTCCGTATGCGCACCCTTA"
                     /Tm="64.2"
                     /%GC="44.4"
                     /Hairpin_Tm="36.7"
                     /Self_Dimer_Tm="14.9"
                     /created_by="slopez"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pSCL317"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oSCL275
                     (<a href=""urn:local:.:vv-ekqhjus"">pSCL317</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oSCL275 (<a href=""urn:local:.:vv-ekqhjus"">pSCL317</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oSCL275 (<a
                     href=""urn:local:.:at-ep1jopm"">pSCL317_reeeee</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oSCL275 (<a
                     href=""urn:local:.:h5-eoii18r"">pSCL317_redesigned</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oSCL275 (<a
                     href=""urn:local:.:h5-eoii18r"">pSCL317_redesigned</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oSCL275 (<a
                     href=""urn:local:.:fq-eoihsv7"">o275-o275</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oSCL275 (<a
                     href=""urn:local:.:fq-eoihsv7"">o275-o275</a>)"
                     /label="oSCL275"
     misc_feature    2710..2724
                     /Color="-1"
                     /label="crispey-ext"
     misc_feature    <2725..2805
                     /ApEinfo_fwdcolor="#85dae9"
                     /ApEinfo_revcolor="#85dae9"
                     /Transferred_From="CRISPEY_retron-guide"
                     /Transferred_Similarity="100.00%"
                     /label="msr"
     misc_feature    2725..2736
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /Transferred_From="CRISPEY_retron-guide"
                     /Transferred_Similarity="100.00%"
                     /label="inverted repeat"
     misc_RNA        2795..>2817
                     /created_by="slopez"
                     /modified_by="slopez"
                     /label="msd"
     donor           2818..2917
                     /created_by="slopez"
                     /label="donor"
     grna            2853..2870
                     /created_by="slopez"
                     /label="faa1 gRNA"
     mutation        complement(2871..2873)
                     /created_by="slopez"
                     /modified_by="slopez"
                     /label="change"
     misc_RNA        <2918..2938
                     /created_by="slopez"
                     /modified_by="slopez"
                     /label="msd"
     CDS             2938
                     /created_by="slopez"
                     /label="RT start"
     misc_feature    complement(2940..2951)
                     /ApEinfo_fwdcolor="#b1ff67"
                     /ApEinfo_revcolor="#b1ff67"
                     /Transferred_From="CRISPEY_retron-guide"
                     /Transferred_Similarity="100.00%"
                     /label="inverted repeat"
     misc_feature    complement(2952..2966)
                     /Color="-1"
                     /label="long primer"
     grna            2967..2984
                     /created_by="slopez"
                     /label="faa1 gRNA"
     misc_feature    2985..3075
                     /ApEinfo_fwdcolor="#7ea7d8"
                     /ApEinfo_revcolor="#7ea7d8"
                     /label="modified SpCas9 scaffold"
     primer_bind     complement(3076..3094)
                     /created_by="slopez"
                     /Sequence="TGGGACCATGCCGGCCATC"
                     /Tm="65.0"
                     /%GC="68.4"
                     /Hairpin_Tm="39.0"
                     /Self_Dimer_Tm="26.8"
                     /modified_by="slopez"
                     /label="oSCL316"
     misc_feature    3076..3146
                     /label="HDV-Ribozyme"
     primer_bind     3076..3094
                     /Sequence="GATGGCCGGCATGGTCCCA"
                     /Tm="65.0"
                     /%GC="68.4"
                     /Hairpin_Tm="44.0"
                     /Self_Dimer_Tm="30.9"
                     /created_by="slopez"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="452-293"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL452
                     (<a href=""urn:local:.:jp-d559yd8"">452-293</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL452 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL452 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL452 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL452 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL452 (<a href=""urn:local:.:6r-d558dj1"">452-294</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL452 (<a href=""urn:local:.:d8-d559d8i"">455-456</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL452 (<a href=""urn:local:.:v-d4vdlzu"">pSCL2</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL452 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL452 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /label="SCL452"
     terminator      3147..3505
                     /label="tGal7"
     primer_bind     3147..3174
                     /Sequence="AAAGAAAGTGGAATATTCATTCATATCA"
                     /Tm="56.1"
                     /%GC="25.0"
                     /Hairpin_Tm="42.1"
                     /Self_Dimer_Tm="17.1"
                     /created_by="slopez"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="922-923 (wt)"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL168
                     (<a href=""urn:local:.:ia-dv5iaof"">922-923 (wt)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL168 (<a href=""urn:local:.:w3-dv5kwc1"">HR cassette (wt
                     primer)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL168 (<a href=""urn:local:.:7d-duuvir9"">pSCL27</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL168 (<a href=""urn:local:.:gc-dvb0jpa"">pSCL204</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL168 (<a href=""urn:local:.:30j-c3qkjq8"">pZS165</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL168 (<a href=""urn:local:.:10l-ckanc0u"">167-168</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL168 (<a href=""urn:local:.:3y-d1qf8q0"">pSCL27</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL168 (<a href=""urn:local:.:3we-d5qx0ep"">167-168</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL168 (<a href=""urn:local:.:3z6-d5qxtf9"">pSCL78</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL168 (<a href=""urn:local:.:12m-cc0z43x"">150-151</a>)"
                     /label="SCL168"
     primer_bind     complement(3224..3243)
                     /%GC="50.0"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Tm="58.2"
                     /created_by="primer3"
                     /Sequence="TTGTCACTCCGTTCAAGTCG"
                     /Mismatches=0
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="pSCL24_alt"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL057
                     (<a href=""urn:local:.:16-cjzjbu1"">pSCL24_alt</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL057 (<a
                     href=""urn:local:.:4i-cwqssks"">pSCL24_alt</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL057 (<a href=""urn:local:.:10p-d55cf47"">pSCL70</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL057 (<a href=""urn:local:.:10p-d55cf47"">pSCL70</a>)"
                     /note="Derived using Geneious Prime 2022.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pSCL78"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL057
                     (<a href=""urn:local:.:3z6-d5qxtf9"">pSCL78</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL057 (<a href=""urn:local:.:a1-ekqc2qq"">pSCL315</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL057 (<a href=""urn:local:.:10p-d55cf47"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL057 (<a href=""urn:local:.:10p-d55cf47"">pSCL70</a>)"
                     /label="SCL057"
     primer_bind     complement(3511..3535)
                     /created_by="slopez"
                     /Sequence="ATACGACTCACTATAGGGCGAATTG"
                     /Tm_(binding_region)="60.3"
                     /Tm_(with_extension)="66.8"
                     /%GC_(binding_region)="44.0"
                     /%GC_(with_extension)="46.7"
                     /Hairpin_Tm_(with_extension)="37.6"
                     /Self_Dimer_Tm_(with_extension)="4.4"
                     /Extension="GTTTCGGTGATGACGGTGAA"
                     /modified_by="slopez"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="452-293"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL456
                     (<a href=""urn:local:.:jp-d559yd8"">452-293</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL456 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL456 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL456 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL456 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL456 (<a href=""urn:local:.:6r-d558dj1"">452-294</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL456 (<a href=""urn:local:.:d8-d559d8i"">455-456</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL456 (<a href=""urn:local:.:v-d4vdlzu"">pSCL2</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL456 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL456 (<a href=""urn:local:.:ux-d55adkc"">452-293 -
                     453-454 - 455-456 - 294-451</a>)"
                     /label="SCL456"
     promoter        complement(3519..3537)
                     /label="T7_promoter"
     misc_feature    3549..3563
                     /label="lacZ_a"
     CDS             3564
                     /label="lacZα-Broken"
     CDS             3565..3925
                     /label="lacZα"
     rep_origin      3708..4163
                     /snapgene_direction="RIGHT"
                     /label="f1 ori"
     CDS             complement(4294..5097)
                     /label="URA3"
     promoter        complement(5098..5318)
                     /label="URA3 promoter"
     primer_bind     complement(5467..5487)
                     /created_by="slopez"
                     /Sequence="AACCTCTGACACATGCAGCTC"
                     /Tm="60.6"
                     /%GC="52.4"
                     /Hairpin_Tm="33.5"
                     /Self_Dimer_Tm="None"
                     /modified_by="slopez"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="293-294 - 295-296"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL293
                     (<a href=""urn:local:.:s-copn8jr"">293-294 - 295-296</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL293 (<a href=""urn:local:.:jp-d559yd8"">452-293</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL293 (<a href=""urn:local:.:7v-cn66apn"">pSCL33</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL293 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL293 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL293 (<a href=""urn:local:.:5p-cn64u0h"">pSCL15</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL293 (<a href=""urn:local:.:8k-d4asw2j"">Vv_ecoli_opt in
                     pET-24(+)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL293 (<a href=""urn:local:.:6r-d558dj1"">452-294</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL293 (<a href=""urn:local:.:7a-cn669je"">293-294</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL293 (<a
                     href=""urn:local:.:8f-d4as1mt"">Vv_proteins_mamm_opt in
                     pET-24(+)</a>)"
                     /label="SCL293"
     primer_bind     5488..5508
                     /created_by="slopez"
                     /Sequence="TTCACCGTCATCACCGAAACG"
                     /Tm="61.2"
                     /%GC="52.4"
                     /Hairpin_Tm="35.0"
                     /Self_Dimer_Tm="None"
                     /modified_by="slopez"
                     /X-Transferred_From="293-294 - 295-296"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL294
                     (<a href=""urn:local:.:s-copn8jr"">293-294 - 295-296</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:7v-cn66apn"">pSCL33</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:5p-cn64u0h"">pSCL15</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:8k-d4asw2j"">Vv_ecoli_opt in
                     pET-24(+)</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:7a-cn669je"">293-294</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a
                     href=""urn:local:.:8f-d4as1mt"">Vv_proteins_mamm_opt in
                     pET-24(+)</a>)"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="294-451 - 453-454 - 455-456 - 452-293"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; SCL294
                     (<a href=""urn:local:.:u0-d55aa9h"">294-451 - 453-454 -
                     455-456 - 452-293</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:s-copn8jr"">293-294 -
                     295-296</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:6j-d558buk"">294-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:7v-cn66apn"">pSCL33</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:fm-d559pe1"">452-294 -
                     453-454 - 455-456 - 293-451</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:w7-d55aimz"">pSCL70</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:5p-cn64u0h"">pSCL15</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:8k-d4asw2j"">Vv_ecoli_opt in
                     pET-24(+)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     SCL294 (<a href=""urn:local:.:6r-d558dj1"">452-294</a>)"
                     /label="SCL294"
ORIGIN      
        1 ttttctttaa tttctttttt tactttctat ttttaattta tatatttata ttaaaaaatt
       61 taaattataa ttatttttat agcacgtgat gaaaaggacc caggtggcac ttttcgggga
      121 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      181 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      241 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      301 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
      361 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
      421 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
      481 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
      541 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
      601 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
      661 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
      721 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
      781 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
      841 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
      901 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      961 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     1021 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1081 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     1141 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     1201 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     1261 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     1321 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     1381 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     1441 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     1501 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     1561 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     1621 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     1681 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     1741 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     1801 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     1861 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcaagt agagggggta
     1921 atttttcccc tttattttgt tcatacattc ttaaattgct ttgcctctcc ttttggaaag
     1981 ctatacttcg gagcactgtt gagcgaaggc tcattagata tattttctgt cattttcctt
     2041 aacccaaaaa taagggaaag ggtccaaaaa gcgctcggac aactgttgac cgtgatccga
     2101 aggactggct atacagtgtt cacaaaatag ccaagctgaa aataatgtgt agctatgttc
     2161 agttagtttg gctagcaaag atataaaagc aggtcggaaa tatttatggg cattattatg
     2221 cagagcatca acatgataaa aaaaaacagt tgaatattcc ctcaaaaggg tgcgcatctg
     2281 atgagtccgt gaggacgaaa cgagctagct cgtcatgata agattccgta tgcgcaccct
     2341 tagcgagagg tttatcatta aggtcaacct ctggatgttg tttcggcatc ctgcattgaa
     2401 tctgagttac tgtctgtttt ccttggaaat gttctattta gaaacagggg aattgcttat
     2461 taacgaaatt gcctgaaggc ctcacaactc tggacattat accattgatg cttgcgtcac
     2521 ttcaggaaac ccgtttcttc tgacgtaagg gtgcgcatac ggaatcttat caattaacga
     2581 aattgcccca gtttcagagc tatgctggaa acagcatagc aagttgaaat aaggctagtc
     2641 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tgttcactgc cgtataggca
     2701 gctaagaaat gataagattc cgtatgcgca cccttagcga gaggtttatc attaaggtca
     2761 acctctggat gttgtttcgg catcctgcat tgaatctgag ttactgtctg ttttcctcag
     2821 gattctttac ctagcttcaa gatgtcgtca aagctaaagg tcttgatatc tcatctaact
     2881 tctttaattc tgttgatggc atcatgagca gattgatagg aaacccgttt cttctgacgt
     2941 aagggtgcgc atacggaatc ttatcagcta aaggtcttga tatcgtttca gagctatgct
     3001 ggaaacagca tagcaagttg aaataaggct agtccgttat caacttgaaa aagtggcacc
     3061 gagtcggtgc tttttgatgg ccggcatggt cccagcctcc tcgctggcgc cggctgggca
     3121 acaccttcgg gtggcgaatg ggacttaaag aaagtggaat attcattcat atcatatttt
     3181 ttctattaac tgcctggttt cttttaaatt ttttattggt tgtcgacttg aacggagtga
     3241 caatatatat atatatatat ttaataatga catcattatc tgtaaatctg attcttaatg
     3301 ctattctagt tatgtaagag tggtcctttc cataaaaaaa aaaaaaaaga aaaaagaatt
     3361 ttaggaatac aatgcagctt gtaagtaaaa tctggaatat tcatatcgcc acaacttctt
     3421 atgcttataa aagcactaat gcctgaattt atgttgaaaa tatgtgtcac aaataaagaa
     3481 actgtgacat ctgacacatt tccacgtacc caattcgccc tatagtgagt cgtattacgc
     3541 gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
     3601 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
     3661 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag
     3721 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3781 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3841 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3901 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3961 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4021 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4081 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4141 caaaatatta acgtttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     4201 atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt
     4261 tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa
     4321 atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt
     4381 tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca catcatccac
     4441 ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat
     4501 aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat
     4561 aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga
     4621 tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac
     4681 ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt
     4741 cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt
     4801 attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa
     4861 tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt
     4921 ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt
     4981 acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt
     5041 ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat
     5101 ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt
     5161 tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct
     5221 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaagaa accgaaatca
     5281 aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaaa gctgtggtat ggtgcactct
     5341 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5401 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5461 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5521 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagga
     5581 cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt taatgatgga ataatttggg
     5641 aatttactct gtgtttattt atttttatgt tttgtatttg gattttagaa agtaaataaa
     5701 gaaggtagaa gagttacgga atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa
     5761 caaaaagcgt actttacata tatatttatt agacaagaaa agcagattaa atagatatac
     5821 attcgattaa cgataagtaa aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac
     5881 agacaagatg aaacaattcg gcattaatac ctgagagcag gaagagcaag ataaaaggta
     5941 gtatttgttg gcgatccccc tagagtcttt tacatcttcg gaaaacaaaa actatt
//
