LOCUS       sequence-440827-       10953 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the gRNA targeting the control site in the human ZEB2 locus 
            for the CRISPRd experiment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10953)
  AUTHORS   Matsui S, Granitto M, Buckley M, Ludwig K, Koigi S, Shiley J, 
            Zacharias WJ, Mayhew CN, Lim HW, Iwafuchi M
  TITLE     Pioneer and PRDM transcription factors coordinate bivalent 
            epigenetic states to safeguard cell fate.
  JOURNAL   Mol Cell. 2024 Jan 4:S1097-2765(23)01026-2. doi: 
            10.1016/j.molcel.2023.12.007.
  PUBMED    38211589
REFERENCE   2  (bases 1 to 10953)
  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: "Mol Cell. 
            2024 Jan 4:S1097-2765(23)01026-2. doi: 
            10.1016/j.molcel.2023.12.007."
COMMENT     Sequence Label: FgH1tUTG-sgRNA.Control-ZEB2_CRISPRd
FEATURES             Location/Qualifiers
     source          1..10953
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(156..175)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     503..524
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    609..1197
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(662..682)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1080..1091)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             1707..1887
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(1913..1930)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1919..2143
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2189..2617
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2276..2295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2486..2507
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2626..2646)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2631..2960
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2811..2946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2873..2892
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3008..3055
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3074..3448
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3578..3699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3615..3634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3669..3688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3748..3764)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3748..3764)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3761..3783)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3772..3788
                     /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)."
     promoter        complement(join(3796..3802,3803..3820,3821..3826))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3841..3862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3979..3996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4150..4738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4230..4249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4909..5700,5701..5769))
                     /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     5532..5551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5770..5874)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5949..5968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6140..6519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6521..6719
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6674..6694
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             6737..6917
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6964..7089
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7582..7815
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8000..8044
                     /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"
     misc_feature    8342..8459
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     8650..8669
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    8714..8732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        8759..8834
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8870..10081
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     9918..9937
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             10119..10739
                     /codon_start=1
                     /gene="synthetic gene"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     CDS             10749..10802
                     /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(10803..10805,10806..10808,10809..10953,1..569)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(10803..10805,10806..10808,10809..10953,1..566)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(10848..10869)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
       61 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      121 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      181 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      241 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
      301 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
      361 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      421 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
      481 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
      541 ctctcggcat ggacgagctg tacaagtaaa gcggccgcga tctacaattc gatatcaagc
      601 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
      661 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
      721 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
      781 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
      841 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
      901 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
      961 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     1021 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     1081 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     1141 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     1201 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     1261 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     1321 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     1381 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
     1441 accacacaca aggctacttc cctgattggc agaactacac accagggcca gggatcagat
     1501 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag
     1561 aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg
     1621 acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg
     1681 cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg
     1741 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     1801 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     1861 tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact
     1921 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1981 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     2041 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     2101 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2161 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2221 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2281 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2341 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2401 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2461 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2521 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2581 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2641 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2701 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2761 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2821 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2881 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2941 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3001 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3061 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     3121 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     3181 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     3241 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     3301 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     3361 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     3421 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     3481 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     3541 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     3601 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3661 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     3721 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3781 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3841 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3901 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3961 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4021 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4081 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4141 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4201 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4261 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4321 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4381 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4441 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4501 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4561 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4621 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4681 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4741 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4801 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4861 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4921 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4981 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5041 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5101 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5161 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5221 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5281 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5341 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5401 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5461 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5521 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5581 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5641 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5701 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5761 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5821 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5881 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     5941 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     6001 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     6061 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     6121 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     6181 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     6241 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     6301 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     6361 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     6421 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     6481 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     6541 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     6601 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     6661 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc
     6721 gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     6781 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     6841 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     6901 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
     6961 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
     7021 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
     7081 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
     7141 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
     7201 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
     7261 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
     7321 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
     7381 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
     7441 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
     7501 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
     7561 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
     7621 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
     7681 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
     7741 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
     7801 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
     7861 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
     7921 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
     7981 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
     8041 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
     8101 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
     8161 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
     8221 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
     8281 agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca
     8341 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     8401 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     8461 gggtttatta cagggacagc agagatccag tttggttaat taaccatgga attcgaacgc
     8521 tgacgtcatc aacccgctcc aaggaatcgc gggcccagtg tcactaggcg ggaacaccca
     8581 gcgcgcgtgc gccctggcag gaagatggct gtgagggaca ggggagtggc gccctgcaat
     8641 atttgcatgt cgctatgtgt tctgggaaat caccataaac gtgaaatgtc tttggatttg
     8701 ggaatcttat aagtccctat cagtgataga gattcccgtc agctaacggt gcagctacgt
     8761 tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg
     8821 caccgagtcg gtgctttttt tttaattaac ccgtgtcggc tccagatctg gcctccgcgc
     8881 cgggttttgg cgcctcccgc gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac
     8941 gaagggcgca gcgagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc
     9001 tcataagact cggccttaga accccagtat cagcagaagg acattttagg acgggacttg
     9061 ggtgactcta gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc
     9121 cttctcggcg attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag
     9181 gacgcgccgg gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg
     9241 tttgtggatc gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt
     9301 tcgtggccgc cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta
     9361 gtctgggtcc gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg
     9421 cggctgttcc cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa
     9481 caaggtgggg ggcatggtgg gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg
     9541 gaaagctctt attcgggtga gatgggctgg ggcaccatct ggggaccctg acgtgaagtt
     9601 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     9661 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     9721 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     9781 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     9841 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     9901 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     9961 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
    10021 tcagtgttag actagtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
    10081 cgaagcttgg gctgcaggtc gactctagag gatccaccat gtccagactg gacaagagca
    10141 aagtcatcaa ctctgccttg gagctcctga atgaagttgg cattgagggc ttgaccacca
    10201 ggaagctggc ccagaagctg ggtgtggagc agcctaccct gtactggcat gtgaagaaca
    10261 agagggctct gcttgatgcc ctggccattg agatgttgga caggcaccac acccacttct
    10321 gccctctgga aggggagtcc tggcaggact tcctgaggaa caatgccaag agcttcagat
    10381 gtgccttgct ctcccaccgg gatggtgcca aagttcactt gggcaccagg cctacagaga
    10441 agcagtatga gaccctggag aaccagctgg cattcctgtg ccaacaaggc ttctccctgg
    10501 aaaatgcctt gtatgccctc tctgctgtgg gccacttcac cttgggctgt gtgctggagg
    10561 accaggagca ccaagttgcc aaggaggaga gggagacccc caccactgac tccatgccac
    10621 cactgctgcg gcaagctatt gagttgtttg accaccaagg ggctgagcct gcattccttt
    10681 ttggcctgga actgatcatc tgtggcctgg aaaagcagct gaaatgtgag tctggctctg
    10741 ggtccggtga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc
    10801 caatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
    10861 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
    10921 acggcaagct gaccctgaag ttcatctgca cca
//
