LOCUS       sequence-437552-       10242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To generate LIG4 KO in human cells. Co-expresses eSpCas9(1.1), GFP 
            and a guide RNA against LIG4 exon 3..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10242)
  AUTHORS   Bossaert M, Moreno A, Peixoto A, Pillaire MJ, Chanut P, Frit P, 
            Calsou P, Loparo JJ, Britton S
  TITLE     Identification of the main barriers to Ku accumulation in chromatin.
  JOURNAL   bioRxiv [Preprint]. 2024 Jan 4:2024.01.03.574002. doi: 
            10.1101/2024.01.03.574002.
  PUBMED    38260538
REFERENCE   2  (bases 1 to 10242)
  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: "bioRxiv 
            [Preprint]. 2024 Jan 4:2024.01.03.574002. doi: 
            10.1101/2024.01.03.574002."
COMMENT     Sequence Label: pCAG-eSpCas9-2A-GFP-sgRNA-Lig4
FEATURES             Location/Qualifiers
     source          1..10242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2269..2316
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2332..2385
                     /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"
     CDS             join(2386..2388,2389..3099)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2386..2388,2389..3099)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2689..2708)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3036..3057
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3127..3144)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3133..3340
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3349..3489
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3349..3478
                     /label=AAV2 ITR
     rep_origin      3564..4019
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3651..3670)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3861..3882
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4036..4055)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4155..4177
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4215..4233)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4301..4405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4406..4474,4475..5266)
                     /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(4624..4643)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5437..6025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5926..5945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        6087..6335
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6087..6107
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6258..6277
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6356..6431
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        6611..6914
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          7193..8209
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     8133..8155
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     8217..8236
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      8284..8293
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8293..8358
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8365..8385
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(8410..10242,1..705,706..708,709..1170,1171..1173,
                     1174..1341,1342..1344,1345..2268)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Slaymaker et al., 2016)"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
       61 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
      121 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
      181 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
      241 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
      301 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
      361 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
      421 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
      481 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
      541 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
      601 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
      661 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctggccga cgactccatc
      721 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
      781 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
      841 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
      901 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
      961 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
     1021 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
     1081 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
     1141 gtgggaaccg ccctgatcaa aaagtaccct gcgctggaaa gcgagttcgt gtacggcgac
     1201 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
     1261 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
     1321 gccaacggcg agatccggaa ggcgcctctg atcgagacaa acggcgaaac cggggagatc
     1381 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
     1441 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
     1501 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
     1561 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
     1621 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
     1681 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
     1741 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
     1801 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
     1861 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
     1921 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
     1981 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
     2041 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
     2101 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
     2161 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
     2221 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
     2281 gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat tcggcagtgg agagggcaga
     2341 ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag caagggcgag
     2401 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2461 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2521 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     2581 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     2641 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2701 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2761 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2821 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     2881 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2941 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3001 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3061 gccgccggga tcactctcgg catggacgag ctgtacaagg aattctaact agagctcgct
     3121 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3181 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3241 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3301 agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag
     3361 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3421 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3481 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3541 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3601 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     3661 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3721 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     3781 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3841 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     3901 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     3961 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4021 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4081 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4141 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4201 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4261 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4321 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4381 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4441 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4501 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4561 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4621 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4681 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4741 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4801 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4861 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4921 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4981 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5041 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5101 atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa
     5161 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5221 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5281 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5341 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5401 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5461 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5521 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5581 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5641 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5701 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5761 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5821 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5881 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5941 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6001 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6061 cttttgctgg ccttttgctc acatgtgagg gcctatttcc catgattcct tcatatttgc
     6121 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
     6181 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     6241 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     6301 tggctttata tatcttgtgg aaaggacgaa acaccggggt aagagaacct tcagtgtttt
     6361 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     6421 cgagtcggtg cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     6481 ttttagcgcg tgcgccaatt ctgcagacaa atggctctag taacggccgc cagtgtgctg
     6541 gaattcaggc tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     6601 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6661 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6721 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6781 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     6841 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6901 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
     6961 cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg
     7021 cggggggggg ggggggggcg gggagggggg gggggggggg gggggggggc gggggggggc
     7081 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat
     7141 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc
     7201 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct
     7261 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg
     7321 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga
     7381 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt
     7441 gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg
     7501 cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc
     7561 cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg
     7621 ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc
     7681 ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg
     7741 gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc
     7801 tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg
     7861 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact
     7921 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag
     7981 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg
     8041 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga
     8101 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg
     8161 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
     8221 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctgaatt ctgcagatcc
     8281 ggtgccacca tggactataa ggaccacgac ggagactaca aggatcatga tattgattac
     8341 aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc
     8401 ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg
     8461 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
     8521 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
     8581 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
     8641 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
     8701 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
     8761 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
     8821 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     8881 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     8941 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     9001 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     9061 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     9121 ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     9181 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     9241 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     9301 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     9361 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     9421 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     9481 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     9541 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     9601 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     9661 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     9721 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     9781 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     9841 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     9901 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     9961 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
    10021 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
    10081 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
    10141 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
    10201 aaaattatca aggacaagga cttcctggac aatgaggaaa ac
//
