LOCUS       sequence-216019-        8932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for inducible expression of eGFP-RNF8 FHA domain 
            fusion; 53BP1-independent recruitment to damaged chromatin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8932)
  AUTHORS   Noordermeer SM, Adam S, Setiaputra D, Barazas M, Pettitt SJ, Ling 
            AK, Olivieri M, Alvarez-Quilon A, Moatti N, Zimmermann M, Annunziato
            S, Krastev DB, Song F, Brandsma I, Frankum J, Brough R, Sherker A, 
            Landry S, Szilard RK, Munro MM, McEwan A, Goullet de Rugy T, Lin ZY,
            Hart T, Moffat J, Gingras AC, Martin A, van Attikum H, Jonkers J, 
            Lord CJ, Rottenberg S, Durocher D
  TITLE     The shieldin complex mediates 53BP1-dependent DNA repair.
  JOURNAL   Nature. 2018 Aug;560(7716):117-121. doi: 10.1038/s41586-018-0340-7. 
            Epub 2018 Jul 18.
  PUBMED    30022168
REFERENCE   2  (bases 1 to 8932)
  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"; doi: 
            "10.1038/s41586-018-0340-7"; journalName: "Nature"; date: "2018-08";
            volume: "560"; issue: "7716"; pages: "117-121"
COMMENT     Sequence Label: pCW-eGFP-FHA
FEATURES             Location/Qualifiers
     source          1..8932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    43..160
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        214..528
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    222..240
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    258..276
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    293..311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    329..347
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    365..383
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    400..418
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    436..454
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     501..525
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    549..573
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(591..593,594..596,597..1307)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(591..593,594..596,597..1307)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(636..657)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(897..916)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1244..1265
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1807..1831)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1849..1866
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1876..2386
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2305..2323
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             2396..2992
                     /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(2396..2415)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2892..2912
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2993..3046
                     /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(3047..3664,3665..3670,3671..3706,3707..3709,3710..3745,
                     3746..3748,3749..3784,3785..3793)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(3833..3853)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4251..4262)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4467..4700
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4772..4893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4809..4828)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4863..4882
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4933..5068
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5068..5139
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5140..5208,5209..6000)
                     /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(5358..5377)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6171..6759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6660..6679
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6913..6930
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7047..7068
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7083..7088,7089..7106,7107..7113)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7121..7137
                     /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     7126..7148
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7145..7161
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7145..7161
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7180..7200
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7182..7200
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7228..7454
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7455..7635
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7682..7807
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8300..8533
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8718..8762
                     /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"
ORIGIN
        1 acaattctcg acctcgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
       61 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
      121 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
      181 cagagatcca ctttggccgc gaatcgatat gtcgagttta ctccctatca gtgatagaga
      241 acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt actccctatc
      301 agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg tatgtcgagt
      361 ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt gatagagaac
      421 gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag gcgtgtacgg
      481 tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagaattggc
      541 tagcatcaac aagtttgtac aaaaaagcag gcttcgaagg agatagaacc atggtgagca
      601 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      661 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      721 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      781 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      841 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      901 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      961 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1021 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     1081 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     1141 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1201 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1261 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc gcgccagccg
     1321 gagccgggga gcccggcttc ttcgtcacag gagaccgcgc cggtggccgg agctggtgcc
     1381 tgcggcgggt ggggatgagc gccgggtggc tgctgctgga agatgggtgc gaggtgactg
     1441 taggacgagg atttggtgtc acataccaac tggtatcaaa aatctgcccc ctgatgattt
     1501 ctcgaaacca ctgtgttttg aagcagaatc ctgagggcca atggacaatt atggacaaca
     1561 agagtctaaa tggtgtttgg ctgaacagag cgcgtctgga acctttaagg gtctattcca
     1621 ttcatcaggg agactacatc caacttggag tgcctctgga aaataaggag aatgcggagt
     1681 atgaatatga agttactgaa gaagactggg agacaatata tccttgtctt tccccaaaga
     1741 atgaccaaat gatagaaaaa aataaggaat tgagaactaa aaggaaattc agtttggatg
     1801 aataggaccc agctttcttg tacaaagtgg tttagtaatg aaccggtcca ccaccaccac
     1861 caccactaag gatccggggt tggggttgcg ccttttccaa ggcagccctg ggtttgcgca
     1921 gggacgcggc tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg
     1981 cacattcttc acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc
     2041 cccggcgacg cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg
     2101 ccggacgtga caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca
     2161 gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcagg
     2221 gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt
     2281 gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc
     2341 ggcagtcggc tccctcgttg accgaatcac cgacctctct ccccagcaat tcaccatgac
     2401 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     2461 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     2521 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     2581 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     2641 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     2701 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     2761 gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     2821 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     2881 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     2941 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg ccgaaggtag
     3001 aggttctctc ctcacttgtg gtgatgttga agaaaaccct ggtccaatgt ctagactgga
     3061 caagagcaaa gtcataaacg gagctctgga attactcaat ggtgtcggta tcgaaggcct
     3121 gacgacaagg aaactcgctc aaaagctggg agttgagcag cctaccctgt actggcacgt
     3181 gaagaacaag cgggccctgc tcgatgccct gccaatcgag atgctggaca ggcatcatac
     3241 ccacttctgc cccctggaag gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc
     3301 ataccgctgt gctctcctct cacatcgcga cggggctaaa gtgcatctcg gcacccgccc
     3361 aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt
     3421 ctccctggag aacgcactgt acgctctgtc cgccgtgggc cactttacac tgggctgcgt
     3481 attggaggaa caggagcatc aagtagcaaa agaggaaaga gagacaccta ccaccgattc
     3541 tatgccccca cttctgagac aagcaattga gctgttcgac cggcagggag ccgaacctgc
     3601 cttccttttc ggcctggaac taatcatatg tggcctggag aaacagctaa agtgcgaaag
     3661 cggcgggccg accgacgccc ttgacgattt tgacttagac atgctcccag ccgatgccct
     3721 tgacgacttt gaccttgata tgctgcctgc tgacgctctt gacgattttg accttgacat
     3781 gctccccggg taactaagta aggatcgatc caagatatcg tattcttaac tatgttgctc
     3841 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3901 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3961 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4021 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4081 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4141 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     4201 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4261 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4321 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct gtttcgcctc
     4381 ggcgtccgga ctagaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     4441 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     4501 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     4561 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     4621 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     4681 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     4741 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     4801 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4861 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     4921 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4981 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5041 aagtagtgag gaggcttttt tggaggcctt tcaaatatgt atccgctcat gagacaataa
     5101 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5161 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5221 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5281 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5341 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5401 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5461 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5521 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5581 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5641 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5701 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5761 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5821 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5881 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5941 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6001 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6061 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6121 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6181 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6241 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6301 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6361 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6421 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6481 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6541 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6601 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6661 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6721 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6781 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6841 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6901 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6961 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7021 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7081 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7141 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     7201 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     7261 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     7321 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     7381 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     7441 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     7501 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     7561 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     7621 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     7681 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     7741 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     7801 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     7861 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     7921 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     7981 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     8041 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     8101 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     8161 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     8221 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     8281 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     8341 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     8401 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     8461 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     8521 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     8581 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     8641 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     8701 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     8761 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     8821 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     8881 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg gg
//
