LOCUS       sequence-305300-        9431 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 9431)
  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 9431)
  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..9431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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        1713..1961
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1713..1733
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1884..1903
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1983..2058
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2065..2279
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     2200..2219
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        2308..2383
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2439..2556
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2695..3873
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2926..3864
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3821..3841
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3886..4482
                     /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(3886..3905)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4382..4402
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4492..4545
                     /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      4552..4561
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4558..5268
                     /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(4703..4721)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4971..4990
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5284..5872
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5337..5357)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5755..5766)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5944..6177
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6255..6376
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6292..6311)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6346..6365
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6416..6551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6571..6590)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6572..6590)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6600..6617)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6600..6616)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6609..6631)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6758..7213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6845..6864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7055..7076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7239..7343
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7344..7412,7413..8204)
                     /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(7562..7581)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8375..8963
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8864..8883
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9117..9134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9251..9272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9287..9292,9293..9310,9311..9317)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9325..9341
                     /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     9330..9352
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9349..9365
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9349..9365
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9384..9404
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9386..9404
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggaccca gagagggcct atttcccatg attccttcat
     1741 atttgcatat acgatacaag gctgttagag agataattag aattaatttg actgtaaaca
     1801 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
     1861 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
     1921 atttcttggc tttatatatc ttgtggaaag gacgaaacac cggcctggct ggacagtcaa
     1981 tggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
     2041 gtggcaccga gtcggtgctt ttttgaacgc tgacgtcatc aacccgctcc aaggaatcgc
     2101 gggcccagtg tcactaggcg ggaacaccca gcgcgcgtgc gccctggcag gaagatggct
     2161 gtgagggaca ggggagtggc gccctgcaat atttgcatgt cgctatgtgt tctgggaaat
     2221 caccataaac gtgaaatgtc tttggatttg ggagtcttat aagttctgta tgagaccact
     2281 ctttcccaga ccgccctgag atctggggtt ttagagctag aaatagcaag ttaaaataag
     2341 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctttttta agcttggcgt
     2401 aactagatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     2461 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2521 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2581 gatccacttt ggcgccggct cgagggggcc cgggtgcaaa gatggataaa gttttaaaca
     2641 gagaggaatc tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc
     2701 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     2761 gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     2821 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     2881 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
     2941 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
     3001 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
     3061 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
     3121 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
     3181 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
     3241 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
     3301 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     3361 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     3421 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     3481 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     3541 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     3601 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     3661 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     3721 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     3781 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     3841 caaagttttt ttcttccatt tcaggtgtcg tgacgtacgg ccaccatgac cgagtacaag
     3901 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3961 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     4021 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     4081 gtcgcggacg acggcgccgc cgtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     4141 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     4201 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     4261 gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     4321 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     4381 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4441 gaaggaccgc gcacctggtg catgacccgc aagcccggtg ccggatcggg agagggcaga
     4501 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccaccggt cgccaccatg
     4561 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     4621 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     4681 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     4741 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     4801 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     4861 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     4921 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     4981 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     5041 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     5101 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     5161 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     5221 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaaac gcgttaagtc
     5281 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5341 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5401 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5461 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5521 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5581 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5641 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     5701 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5761 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5821 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
     5881 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     5941 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     6001 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6061 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6121 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     6181 cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6241 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6301 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6361 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     6421 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6481 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6541 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     6601 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     6661 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     6721 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     6781 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     6841 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6901 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     6961 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     7021 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     7081 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     7141 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     7201 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     7261 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     7321 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     7381 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     7441 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     7501 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     7561 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     7621 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     7681 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7741 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7801 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7861 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     7921 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7981 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     8041 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     8101 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     8161 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     8221 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     8281 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     8341 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8401 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8461 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8521 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8581 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8641 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8701 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     8761 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8821 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8881 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8941 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     9001 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     9061 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     9121 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     9181 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     9241 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     9301 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     9361 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     9421 tgcaagctta a
//
