LOCUS       sequence-276484-        9430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-Cas9 Knock down.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9430)
  AUTHORS   Arnan C, Ullrich S, Pulido-Quetglas C, Nurtdinov R, Esteban A, 
            Blanco-Fernandez J, Aparicio-Prat E, Johnson R, Perez-Lluch S, Guigo
            R
  TITLE     Paired guide RNA CRISPR-Cas9 screening for protein-coding genes and 
            lncRNAs involved in transdifferentiation of human B-cells to 
            macrophages.
  JOURNAL   BMC Genomics. 2022 May 26;23(1):402. doi: 
            10.1186/s12864-022-08612-7.
  PUBMED    35619054
REFERENCE   2  (bases 1 to 9430)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC 
            Genomics."; date: "2022-05-26"; volume: "23(1):402. doi"; pages: " 
            10.1186/s12864-022-08612-7"
COMMENT     Sequence Label: pDECKO_mCherry_CD19.4
FEATURES             Location/Qualifiers
     source          1..9430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..1264
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1255
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1212..1232
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1277..1873
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1277..1296)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1773..1793
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1883..1936
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      1943..1952
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1949..2659
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2094..2112)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2362..2381
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2675..3263
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2728..2748)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3146..3157)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3335..3568
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3646..3767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3683..3702)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3737..3756
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3807..3942
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3962..3981)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3963..3981)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3991..4008)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3991..4007)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4000..4022)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4149..4604
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4236..4255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4446..4467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4630..4734
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4735..4803,4804..5595)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4953..4972)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5766..6354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6255..6274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6508..6525
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6642..6663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6678..6683,6684..6701,6702..6708)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6716..6732
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6721..6743
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6740..6756
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6740..6756
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6775..6795
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6777..6795
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6823..7049
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7050..7230
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7277..7402
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7895..8128
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8313..8357
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        8535..8783
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8535..8555
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8706..8725
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8805..8880
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8887..9101
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     9022..9041
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        9130..9205
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    9261..9378
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccgggtgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccactggctg cagtacgtga ttcttgatcc
      421 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
      481 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
      541 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
      601 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
      661 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
      721 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
      781 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
      841 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
      901 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
      961 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1021 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1081 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1141 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1201 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     1261 gtgacgtacg gccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     1321 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     1381 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     1441 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg ccgtggcggt
     1501 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     1561 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     1621 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca
     1681 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     1741 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     1801 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     1861 caagcccggt gccggatcgg gagagggcag aggaagtctg ctaacatgcg gtgacgtcga
     1921 ggagaatcct ggcccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc
     1981 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     2041 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     2101 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     2161 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     2221 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     2281 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     2341 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     2401 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     2461 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     2521 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     2581 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     2641 ggacgagctg tacaagtaaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     2701 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     2761 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2821 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2881 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2941 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     3001 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     3061 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     3121 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     3181 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     3241 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     3301 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     3361 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     3421 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     3481 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     3541 ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt
     3601 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     3661 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3721 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     3781 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3841 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3901 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     3961 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     4021 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4081 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4141 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4201 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4261 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4321 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4381 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4441 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4501 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4561 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     4621 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4681 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4741 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4801 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4861 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4921 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4981 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5041 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5101 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5161 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5221 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5281 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5341 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5401 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5461 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5521 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5581 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5641 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5701 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5761 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5821 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5881 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5941 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6001 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6061 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6121 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6181 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6241 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6301 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6361 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6421 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6481 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6541 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6601 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6661 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     6721 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     6781 aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct tatgcaatac
     6841 tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa ggagagaaaa
     6901 agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta ttaggaaggc
     6961 aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc agagatattg
     7021 tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag atctgagcct
     7081 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     7141 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     7201 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     7261 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     7321 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     7381 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     7441 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     7501 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     7561 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     7621 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     7681 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     7741 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     7801 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     7861 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     7921 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     7981 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     8041 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     8101 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     8161 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     8221 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     8281 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     8341 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     8401 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     8461 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     8521 cgaggggacc cagagagggc ctatttccca tgattccttc atatttgcat atacgataca
     8581 aggctgttag agagataatt agaattaatt tgactgtaaa cacaaagata ttagtacaaa
     8641 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     8701 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttataca
     8761 tcttgtggaa aggacgaaac accggcctgg ctggacagtc aatggtttta gagctagaaa
     8821 tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc
     8881 ttttttgaac gctgacgtca tcaacccgct ccaaggaatc gcgggcccag tgtcactagg
     8941 cgggaacacc cagcgcgcgt gcgccctggc aggaagatgg ctgtgaggga caggggagtg
     9001 gcgccctgca atatttgcat gtcgctatgt gttctgggaa atcaccataa acgtgaaatg
     9061 tctttggatt tgggagtctt ataagttctg tatgagacca ctctttccca gaccgccctg
     9121 agatctgggg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     9181 tgaaaaagtg gcaccgagtc ggtgcttttt taagcttggc gtaactagat cttgagacaa
     9241 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     9301 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     9361 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact ttggcgccgg
     9421 ctcgaggggg
//
