LOCUS       Addgene_p3015b_with_annotations        7740 bp ds-DNA     circular SYN 01-JUL-2024
DEFINITION  Expresses a sgRNA template with a protospacer cloning site flanked
            by BsaI restriction sites for easy insertion of a targeting
            cassette.
ACCESSION
VERSION     .
KEYWORDS    p3015b.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct.
REFERENCE   1  (bases 1 to 7740)
  AUTHORS   Gopalakrishna KP, Hillebrand GH, Bhavana VH, Elder JL, D'Mello A,
            Tettelin H and Hooven TA
  TITLE     Group B Streptococcus Cas9 variants provide insight into
            programmable gene repression and CRISPR-Cas transcriptional
            effects.
  JOURNAL   Commun Biol. 2023 Jun 9;6(1):620. doi: 10.1038/s42003-023-04994-w.
  PUBMED    37296208
REFERENCE   2  (bases 1 to 7740)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 3, 2024 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7740
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      214..270
                     /created_by="User"
                     /X-Transferred_From="3015b Flex sgRNA Deleted Cryptic
                     Promoter"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     Originof Replication (<a
                     href=""urn:local:.:32d-c8zthnm"">3015bFlex sgRNA Deleted
                     Cryptic Promoter</a>)"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="cfbProm-eGFP into 3015-CPF GA"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; Origin
                     of Replication (<a
                     href=""urn:local:.:357-c906sen"">cfbProm-eGFP into
                     3015-CPF GA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Origin of Replication (<a
                     href=""urn:local:.:32d-c8zthnm"">3015b Flex sgRNA Deleted
                     Cryptic Promoter</a>)"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015b_mCherry_SHAM 4_17_24 Edited for
                     Addgene"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; Origin
                     of Replication (<a
                     href=""urn:local:.:yi-hwr7qk0"">3015b_mCherry_SHAM 4_17_24
                     Edited for Addgene</a>)"
                     /label="Origin of Replication"
     promoter        780..843
                     /created_by="User"
                     /X-Transferred_From="3010 Flexible sgRNA"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     xyl/tet Promoter (<a href=""urn:local:.:32f-c8zthnn"">3010
                     Flexible sgRNA</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/-1;
                     xyl/tet Promoter (<a href=""urn:local:.:32c-c8zthnl"">3015
                     Flexible sgRNA</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/-1;
                     xyl/tet Promoter (<a
                     href=""urn:local:.:32e-c8zthnn"">3015b Flex sgRNA + cylE55
                     PS</a>)"
                     /X-Alternative_Match="100.00%; Adjusted-Interval-0:0/-1;
                     xyl/tet Promoter (<a
                     href=""urn:local:.:32d-c8zthnm"">3015b Flex sgRNA Deleted
                     Cryptic Promoter</a>)"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="cfbProm-eGFP into 3015-CPF GA"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; xyl/tet
                     Promoter (<a href=""urn:local:.:357-c906sen"">cfbProm-eGFP
                     into 3015-CPF GA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     xyl/tet Promoter (<a href=""urn:local:.:32f-c8zthnn"">3010
                     Flexible sgRNA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/-1;
                     xyl/tet Promoter (<a href=""urn:local:.:32c-c8zthnl"">3015
                     Flexible sgRNA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/-1;
                     xyl/tet Promoter (<a
                     href=""urn:local:.:32e-c8zthnn"">3015b Flex sgRNA + cylE55
                     PS</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/-1;
                     xyl/tet Promoter (<a
                     href=""urn:local:.:32d-c8zthnm"">3015b Flex sgRNA Deleted
                     Cryptic Promoter</a>)"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015b_mCherry_SHAM 4_17_24 Edited for
                     Addgene"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; xyl/tet
                     Promoter (<a
                     href=""urn:local:.:yi-hwr7qk0"">3015b_mCherry_SHAM 4_17_24
                     Edited for Addgene</a>)"
                     /label="xyl/tet Promoter"
     note            <844..>871
                     /created_by="User"
                     /X-Transferred_From="3010 Flexible sgRNA, Digestion of
                     3010 Flexible sgRNA - Fragment 1"
                     /X-Transferred_Similarity="100.00%"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015 Flexible sgRNA"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     Protospacer Sequence (<a
                     href=""urn:local:.:32c-c8zthnl"">3015 Flexible sgRNA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     Protospacer Sequence (<a
                     href=""urn:local:.:357-c906sen"">cfbProm-eGFP into
                     3015-CPF GA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:1/0;
                     Protospacer Sequence (<a
                     href=""urn:local:.:32f-c8zthnn"">3010 Flexible sgRNA</a>)"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015b_mCherry_SHAM 4_17_24 Edited for
                     Addgene"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     Protospacer Sequence (<a
                     href=""urn:local:.:yi-hwr7qk0"">3015b_mCherry_SHAM 4_17_24
                     Edited for Addgene</a>)"
                     /label="Protospacer Sequence"
     note            844..849
                     /created_by="smc345"
                     /label="BsaI Site to Remove Sham Protospacer"
     note            866..871
                     /created_by="smc345"
                     /label="BsaI Cut Site to Remove Sham Protospacer"
     misc_RNA        <872..975
                     /created_by="User"
                     /modified_by="User"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015 Flexible sgRNA"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; TRACR
                     (<a href=""urn:local:.:32c-c8zthnl"">3015 Flexible
                     sgRNA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     TRACR (<a href=""urn:local:.:357-c906sen"">cfbProm-eGFP
                     into 3015-CPF GA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:1/0;
                     TRACR (<a href=""urn:local:.:32e-c8zthnn"">3015b Flex
                     sgRNA + cylE55 PS</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:1/0;
                     TRACR (<a href=""urn:local:.:32d-c8zthnm"">3015b Flex
                     sgRNA Deleted Cryptic Promoter</a>)"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015b_mCherry_SHAM 4_17_24 Edited for
                     Addgene"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; TRACR
                     (<a href=""urn:local:.:yi-hwr7qk0"">3015b_mCherry_SHAM
                     4_17_24 Edited for Addgene</a>)"
                     /label="TRACR"
     primer_bind     complement(1022..1039)
                     /note="L4440 vector, forward primer"
                     /label="L4440"
     rep_origin      complement(1193..1781)
                     /direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     primer_bind     complement(1273..1292)
                     /note="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     promoter        1960..2049
                     /created_by="User"
                     /X-Transferred_From="3012 BSAI Fix C5, 3012 cylE 30 bp PS
                     + US CR GA Product"
                     /X-Transferred_Similarity="100.00%"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015 Flexible sgRNA"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; TetR
                     Promoter (<a href=""urn:local:.:32c-c8zthnl"">3015
                     Flexible sgRNA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     TetRPromoter (<a href=""urn:local:.:32e-c8zthnn"">3015b
                     FlexsgRNA + cylE55 PS</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     TetRPromoter (<a
                     href=""urn:local:.:357-c906sen"">cfbProm-eGFPinto 3015-CPF
                     GA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     TetRPromoter (<a href=""urn:local:.:32d-c8zthnm"">3015b
                     FlexsgRNA Deleted Cryptic Promoter</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     TetRPromoter (<a
                     href=""urn:local:.:32f-c8zthnn"">3010Flexible sgRNA</a>)"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015b_mCherry_SHAM 4_17_24 Edited for
                     Addgene"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; TetR
                     Promoter (<a
                     href=""urn:local:.:yi-hwr7qk0"">3015b_mCherry_SHAM 4_17_24
                     Edited for Addgene</a>)"
                     /label="TetR Promoter"
     RBS             2034..2042
                     /note="full consensus sequence for ribosome-binding sites
                     upstream of start codons in E. coli; complementary to
                     aregion in the 3' end of the 16S rRNA (Chen et al., 1994)"
                     /label="Shine-Dalgarno sequence"
     gene            2050..2673
                     /created_by="User"
                     /X-Transferred_From="3010 cylE 30 bp sgRNA GA Product,
                     3012_3013 cylE 30 bp sgRNA GA Product"
                     /X-Transferred_Similarity="100.00%"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015 Flexible sgRNA"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; TetR
                     (<ahref=""urn:local:.:32c-c8zthnl"">3015 Flexible
                     sgRNA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     TetR(<a href=""urn:local:.:32e-c8zthnn"">3015b Flex sgRNA
                     +cylE55 PS</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     TetR(<a href=""urn:local:.:357-c906sen"">cfbProm-eGFP
                     into3015-CPF GA</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     TetR(<a href=""urn:local:.:32d-c8zthnm"">3015b Flex
                     sgRNADeleted Cryptic Promoter</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     TetR(<a href=""urn:local:.:32f-c8zthnn"">3010
                     FlexiblesgRNA</a>)"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015b_mCherry_SHAM 4_17_24 Edited for
                     Addgene"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; TetR (<a
                     href=""urn:local:.:yi-hwr7qk0"">3015b_mCherry_SHAM 4_17_24
                     Edited for Addgene</a>)"
                     /label="TetR"
     primer_bind     complement(2123..2142)
                     /note="Tetracycline resistance gene, reverse primer"
                     /label="Tet-R"
     gene            3219..3956
                     /gene="ermAM"
                     /X-Transferred_From="pORI23 (based on high-quality MCS
                     sequencing data)"
                     /X-Transferred_Similarity="100.00%"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pMBsacB"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; ermAM
                     gene (<a href=""urn:local:.:34b-c8zu34j"">pMBsacB</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     ermAM gene (<a href=""urn:local:.:34g-c8zu3ga"">pORI23
                     (based on high-quality MCS sequencing data)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/-4;
                     Erm(<a href=""urn:local:.:34b-c8zu34j"">pMBsacB</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/-4;
                     Erm(<a
                     href=""urn:local:.:33l-c8ztuar"">pDCErm(deduced)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/-4;
                     Erm(<a href=""urn:local:.:33t-c8ztve7"">pHY304
                     GenbankSubmission</a>)"
                     /modified_by="smc345"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015b_mCherry_SHAM 4_17_24 Edited for
                     Addgene"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; ermAM
                     gene (<a
                     href=""urn:local:.:yi-hwr7qk0"">3015b_mCherry_SHAM 4_17_24
                     Edited for Addgene</a>)"
     note            4338..4437
                     /created_by="smc345"
                     /label="Cfb Promoter"
     note            4423..4428
                     /created_by="smc345"
                     /label="Ribosome Binding Site"
     gene            4438..>4457
                     /gene="cfb"
                     /locus_tag="SAK_1983"
                     /db_xref="GeneID:3686873"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pBSU101 cfb-promoter eGFP & downstream"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; cfb
                     gene(<a href=""urn:local:.:338-c8ztrii"">pBSU101
                     cfb-promotereGFP & downstream</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; cfb
                     gene (<a href=""urn:local:.:35m-c94fcjg"">pBSU101
                     cfb-promoter mCherry & downstream</a>)"
                     /modified_by="smc345"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015b_mCherry_SHAM 4_17_24 Edited for
                     Addgene"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; cfb gene
                     fragment (<a
                     href=""urn:local:.:yi-hwr7qk0"">3015b_mCherry_SHAM 4_17_24
                     Edited for Addgene</a>)"
                     /label="cfb gene fragment"
     CDS             4465..5175
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent
                     protein (Shaner et al., 2004)"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVM
                     NFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDG
                     ALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQ
                     YERAEGRHSTGGMDELYK"
                     /label="mCherry"
     primer_bind     complement(4610..4628)
                     /note="mCherry, reverse primer"
                     /label="mCherry-R"
     primer_bind     4878..4897
                     /note="mCherry, forward primer"
                     /label="mCherry-F"
     primer_bind     complement(5209..5226)
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /label="M13 Forward"
     primer_bind     complement(5209..5225)
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /label="M13 fwd"
     primer_bind     complement(5218..5240)
                     /note="In lacZ gene"
                     /label="M13/pUC Forward"
     gene            6121..6420
                     /gene="repD"
                     /note="Derived using Geneious Prime 2020.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pORI23 (based on high-quality MCS
                     sequencing data)"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; repD
                     gene (<a href=""urn:local:.:34g-c8zu3ga"">pORI23 (based on
                     high-quality MCS sequencing data)</a>)"
                     /note="Derived using Geneious Prime 2023.2.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="3015b_mCherry_SHAM 4_17_24 Edited for
                     Addgene"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; repD
                     gene (<a
                     href=""urn:local:.:yi-hwr7qk0"">3015b_mCherry_SHAM 4_17_24
                     Edited for Addgene</a>)"
ORIGIN      
        1 ccaggaagaa atggtggcat tcaacttgct agtgttaaat cattgttgct atcgatcatt
       61 aaagtaaaaa aagaagaaaa agaaagctat ataaaggcgc tgacaaattc ttttgactta
      121 gagcatacat tcattcaaga gactttaaac aagctagcag aacgccctaa aacggacaca
      181 caactcgatt tgtttagcta tgatacaggc tgaaaataaa acccgcacta tgccattaca
      241 tttatatcta tgatacgtgt ttgttttttc tttgctgttt agcgaatgat tagcagaaat
      301 atacagagta agattttaat taattattag ggggagaagg agagagtagc ccgaaaactt
      361 ttagttggct tggactgaac gaagtgaggg aaaggctact aaaacgtcga ggggcagtga
      421 gagcgaagcg aacacttgat tttttaattt tctatctttt ataggtcatt agagtatact
      481 tatttgtcct ataaactatt tagcagcata atagatttat tgaataggtc atttaagttg
      541 agcatattag aggaggaaaa tcttggagaa atatttgaag aacccgatta catggattgg
      601 attagttctt gtggttacgt ggtttttaac taaaagtagt gaatttttga tttttggtgt
      661 gtgtgtcttg ttgttagtat ttgctagtca aagtgattaa atagaattct catgtttgac
      721 agcttatcat cggagctccg atgataagct gtcaaacatg agaattcccg gggatcctgt
      781 cgagttcatg aaaaactaaa aaaaatattg acactctatc attgatagag tataattaaa
      841 atagagaccg cgtactacgc ctattggtct cgttttagag ctgtgaaaca gcacagcacg
      901 ttaaaataag gcagtgattt ttaatccagt ccgtattcag cttgaaaaag tgagcaccga
      961 atcggtgctt tttttctaga gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1021 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1081 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1141 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1201 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1261 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1321 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1381 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1441 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1501 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1561 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1621 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1681 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1741 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1801 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1861 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1921 aaagtatata tgagtaaact tggtctgaca gccctcgagc ataaggctcg taggctatat
     1981 tcagggcgac cacaacggtt tccctctaca aataattttg tttaactttg aaataaggag
     2041 gtaatacaaa tgtctcgttt agataaaagt aaagtgatta acagcgcatt agagctgctt
     2101 aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag
     2161 cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt
     2221 gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat
     2281 tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca
     2341 aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta
     2401 gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg
     2461 gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa
     2521 agggaaacac ctactactga tagtatgccg ccattattac gacaagctat cgaattattt
     2581 gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta
     2641 gaaaaacaac ttaaatgtga aagtgggtct taactcgagc caatcgatac gattttgaag
     2701 tggcaacaga taaaaaaaag cagtttaaaa ttgttgctga acttttaaaa caagcaaata
     2761 caatcattgt cgcaacagat agcgacagag aaggcgaaaa cattgcctgg tcgatcattc
     2821 ataaagcaaa tgccttttct aaagataaaa cgtataaaag actatggatc aatagtttag
     2881 aaaaagatgt gatccgtagc ggttttcaaa atttgcgacc aggaatgaat tactatccct
     2941 tttatcaaga agcgcacaaa aagaaaaacg aaatgataca ccaatcagtg caaaaaaaga
     3001 tataatggga gataagacgg ttcgtgttcg tgctgacttg caccatatca taaaaatcga
     3061 aacagcaaag aatggcggaa acgtaaaaga agttatggaa ataagactta gaagcaaact
     3121 taagagtgtg ttgatagtgc agtatcttaa aattttgtat aataggaatt gaagttaaat
     3181 tagatgctaa aaatttgtaa ttaagaagga gtgattacat gaacaaaaat ataaaatatt
     3241 ctcaaaactt tttaacgagt gaaaaagtac tcaaccaaat aataaaacaa ttgaatttaa
     3301 aagaaaccga taccgtttac gaaattggaa caggtaaagg gcatttaacg acgaaactgg
     3361 ctaaaataag taaacaggta acgtctattg aattagacag tcatctattc aacttatcgt
     3421 cagaaaaatt aaaactgaat actcgtgtca ctttaattca ccaagatatt ctacagtttc
     3481 aattccctaa caaacagagg tataaaattg ttgggagtat tccttaccat ttaagcacac
     3541 aaattattaa aaaagtggtt tttgaaagcc atgcgtctga catctatctg attgttgaag
     3601 aaggattcta caagcgtacc ttggatattc accgaacact agggttgctc ttgcacactc
     3661 aagtctcgat tcagcaattg cttaagctgc cagcggaatg ctttcatcct aaaccaaaag
     3721 taaacagtgt cttaataaaa cttacccgcc ataccacaga tgttccagat aaatattgga
     3781 agctatatac gtactttgtt tcaaaatggg tcaatcgaga atatcgtcaa ctgtttacta
     3841 aaaatcagtt tcatcaagca atgaaacacg ccaaagtaaa caatttaagt accgttactt
     3901 atgagcaagt attgtctatt tttaatagtt atctattatt taacgggagg aaataattct
     3961 atgagtcgct tttgtaaatt tggaaagtta cacgttacta aagggaatgt agataaatta
     4021 ttaggtatac tactgacagc ttccaaggag ctaaagagat ctaaaatagt acgtttctta
     4081 ttaaatttat tttttaagaa aaacattagg ctaacctatt tgataaaaat agtgataagt
     4141 agaaattttc taaaatattt agatttatat aaggtggtaa attatcgtta tggtttttta
     4201 catgaaaatc taaattaaat tatcagaaat taacaaaagg ataggcgcaa gtatattata
     4261 gcaaaatgat aatattagtt ttatgaaaaa aatgcagttt atttgtaata atgtaaaata
     4321 taggggaaaa gaaagcgctt gacgaccttt tggacaagta gtaagataca aacatgggcc
     4381 ctgtaaatta aaaatactgc agtaaaagtg attttagttt aaaggaggaa atttattatg
     4441 aacgttaaac atatgatgga tcccatggtg agcaagggcg aggaggataa catggccatc
     4501 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     4561 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     4621 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     4681 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     4741 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     4801 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     4861 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     4921 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     4981 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     5041 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     5101 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     5161 gagctgtaca agtaatctag agtcgacctg caggcatgca agcttggcac tggccgtcgt
     5221 tttacaacgt cgtgactggg aaaaccctgg cctagcgctt agaatcgctt taggaaacac
     5281 gatccagtcc aataatcgtc gataaaaact tttgaaaaag gttggtgaaa ttacctactt
     5341 ttggaataat cacaaatcac aagtgattaa tcacaaatca caagtgatta atcacttgtt
     5401 tattaagata ttaaaagcta taatttaaat aaagcgtgaa ttttattaca caaaaagagg
     5461 ggggagaaac ttggaactag catttagaga aagcttaaaa aagatgagag gtaccaaatc
     5521 aaaagaaaaa ttctcccaag aattagaaat gagtagatca aattattcac gaatagaatc
     5581 aggaaaatca gatccaacca taaaaacact agaacaaatt gcaaagttaa ctaactcaac
     5641 gctagtagtg gatttaatcc caaatgagcc aacagaacca gaaccagaaa cagaatcaga
     5701 acaagtaaca ttggatttag aaatggaaga agaaaaaagc aatgacttcg tgtgaataat
     5761 gcgcgaaatc gttgcttatt ttttttaaaa gcggtatact agatataacg aaacaacgaa
     5821 ctgaatagaa acgaaaaaag agccatgaca catttataaa atgtttgacg acattttata
     5881 aatgcatagc ccgataagat tgccaaacca acgcttatca gttagtcaga tgaactcttc
     5941 cctcgtaaga agttatttaa ttaactttgt ttgaagacgg tatataaccg tactatcatt
     6001 atatagggaa atcagagagt tttcaagtat ctaagctact gaatttaaga attgttaagc
     6061 aatcaatcgg aaatcgtttg attgcttttt ttgtattcat ttatagaagg tggagtttgt
     6121 atgaatcatg atgaatgtaa aacttatata aaaaatagtt tattggagat aagaaaatta
     6181 gcaaatatct atacactaga aacgtttaag aaagagttag aaaagagaaa tatctactta
     6241 gaaacaaaat cagataagta tttttcttcg gagggggaag attatatata taagttaata
     6301 gaaaataaca aaataattta ttcgattagt ggaaaaaaat tgacttataa aggaaaaaaa
     6361 tctttttcaa aacatgcaat attgaaacag ttgaatgaaa aagcaaacca agttaattaa
     6421 acaacctatt ttataggatt tataggaaag gagaacagct gaatgaatat cccttttgtt
     6481 gtagaaactg tgcttcatga cggcttgtta aagtacaaat ttaaaaatag taaaattcgc
     6541 tcaatcacta ccaagccagg taaaagcaaa ggggctattt ttgcgtatcg ctcaaaatca
     6601 agcatgattg gcggtcgtgg tgttgttctg acttccgagg aagcgattca agaaaatcaa
     6661 gatacattta cacattggac acccaacgtt tatcgttatg gaacgtatgc agacgaaaac
     6721 cgttcataca cgaaaggaca ttctgaaaac aatttaagac aaatcaatac cttctttatt
     6781 gattttgata ttcacacggc aaaagaaact atttcagcaa gcgatatttt aacaaccgct
     6841 attgatttag gttttatgcc tactatgatt atcaaatctg ataaaggtta tcaagcatat
     6901 tttgttttag aaacgccagt ctatgtgact tcaaaatcag aatttaaatc tgtcaaagca
     6961 gccaaaataa tttcgcaaaa tatccgagaa tattttggaa agtctttgcc agttgatcta
     7021 acgtgtaatc attttggtat tgctcgcata ccaagaacgg acaatgtaga attttttgat
     7081 cctaattacc gttattcttt caaagaatgg caagattggt ctttcaaaca aacagataat
     7141 aagggcttta ctcgttcaag tctaacggtt ttaagcggta cagaaggcaa aaaacaagta
     7201 gatgaaccct ggtttaatct cttattgcac gaaacgaaat tttcaggaga aaagggttta
     7261 atagggcgta ataacgtcat gtttaccctc tctttagcct actttagttc aggctattca
     7321 atcgaaacgt gcgaatataa tatgtttgag tttaataatc gattagatca acccttagaa
     7381 gaaaaagaag taatcaaaat tgttagaagt gcctattcag aaaactatca aggggctaat
     7441 agggaataca ttaccattct ttgcaaagct tgggtatcaa gtgatttaac cagtaaagat
     7501 ttatttgtcc gtcaagggtg gtttaaattc aagaaaaaaa gaagcgaacg tcaacgtgtt
     7561 catttgtcag aatggaaaga agatttaatg gcttatatta gcgaaaaaag cgatgtatac
     7621 aagccttatt tagtgacgac caaaaaagag attagagaag tgctaggcat tcctgaacgg
     7681 acattagata aattgctgaa ggtactgaag gcgaatcagg aaattttctt taagattaaa
//
