LOCUS       sequence-420407-        9551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA-2 against ZWILCH.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9551)
  AUTHORS   Brambati A, Sacco O, Porcella S, Heyza J, Kareh M, Schmidt JC, Sfeir
            A
  TITLE     RHINO directs MMEJ to repair DNA breaks in mitosis.
  JOURNAL   Science. 2023 Aug 11;381(6658):653-660. doi: 
            10.1126/science.adh3694. Epub 2023 Jul 13.
  PUBMED    37440612
REFERENCE   2  (bases 1 to 9551)
  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: 
            "10.1126/science.adh3694"; journalName: "Science"; date: 
            "2023-08-11- 11"; volume: "381"; issue: "6658"; pages: "653-660"
COMMENT     Sequence Label: lenti-ZWILCH gRNA2
FEATURES             Location/Qualifiers
     source          1..9551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    282..399
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        450..698
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     450..470
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     621..640
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        720..795
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        857..1068
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1076..1085
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1082..1084,1085..1087,1088..1798)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1082..1084,1085..1087,1088..1798)
                     /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(1127..1148)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1388..1407)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1735..1756
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1799..1819
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1829..1885
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1886..2482
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     2379..2399
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2498..3086
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2551..2571)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2969..2980)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3158..3391
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(3417..3434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3423..3647
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3693..4121
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3780..3799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3990..4011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4130..4150)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4135..4464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4315..4450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4377..4396
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4565..4939
                     /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    5069..5190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5106..5125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5160..5179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5239..5255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5239..5255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5252..5274)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5263..5279
                     /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(5287..5293,5294..5311,5312..5317))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5332..5353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5470..5487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5641..6229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5721..5740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6400..7191,7192..7260))
                     /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     7023..7042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7261..7365)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7440..7459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7631..8010
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8012..8210
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8165..8185
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             8228..8408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8455..8580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9073..9306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9491..9535
                     /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 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
       61 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
      121 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
      181 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc
      241 gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
      301 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
      361 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
      421 agagatccag tttggttaat taaggtaccg agggcctatt tcccatgatt ccttcatatt
      481 tgcatatacg atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa
      541 agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt
      601 taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt
      661 tcttggcttt atatatcttg tggaaaggac gaaacaccgg ttaattggac tttacaccag
      721 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
      781 gcaccgagtc ggtgcttttt tgaattcgct agctaggtct tgaaaggagt gggaattggc
      841 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
      901 ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
      961 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
     1021 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagga ccggtgccac
     1081 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1141 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1201 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1261 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1321 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1381 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1441 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1501 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1561 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1621 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1681 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1741 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagcc
     1801 caagaaaaag cggaaagtgg gatccggcgc aacaaacttc tctctgctga aacaagccgg
     1861 agatgtcgaa gagaatcctg gaccgaccga gtacaagccc acggtgcgcc tcgccacccg
     1921 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     1981 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     2041 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     2101 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     2161 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     2221 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga
     2281 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     2341 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     2401 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     2461 gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat
     2521 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2581 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2641 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2701 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2761 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2821 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2881 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     2941 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3001 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3061 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     3121 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     3181 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     3241 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     3301 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     3361 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
     3421 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3481 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3541 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3601 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3661 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3721 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3781 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3841 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3901 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3961 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4021 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4081 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4141 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4201 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4261 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4321 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4381 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4441 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4501 gatcagcacg tgttgacaat taatcatcgg catagtataa tacgacaagg tgaggaacta
     4561 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     4621 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     4681 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     4741 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     4801 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     4861 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     4921 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     4981 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5041 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5101 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5161 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5221 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5281 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5341 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5401 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5461 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5521 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5581 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5641 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5701 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5761 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5821 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5881 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5941 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6001 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6061 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6121 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6181 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6241 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6301 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6361 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6421 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6481 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6541 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6601 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6661 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6721 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6781 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6841 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6901 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6961 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7021 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7081 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7141 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7201 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7261 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7321 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7381 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     7441 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7501 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     7561 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     7621 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     7681 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     7741 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     7801 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     7861 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     7921 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     7981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     8041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     8101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     8161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc
     8221 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     8281 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     8341 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     8401 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
     8461 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     8521 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     8581 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     8641 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     8701 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     8761 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     8821 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     8881 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
     8941 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     9001 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     9061 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     9121 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     9181 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     9241 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     9301 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     9361 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     9421 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     9481 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     9541 ttggtttaac a
//
