LOCUS       sequence-404973-       11759 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual gRNA plasmid for UCK2 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11759)
  AUTHORS   Girish V, Lakhani AA, Thompson SL, Scaduto CM, Brown LM, Hagenson 
            RA, Sausville EL, Mendelson BE, Kandikuppa PK, Lukow DA, Yuan ML, 
            Stevens EC, Lee SN, Schukken KM, Akalu SM, Vasudevan A, Zou C, 
            Salovska B, Li W, Smith JC, Taylor AM, Martienssen RA, Liu Y, Sun R,
            Sheltzer JM
  TITLE     Oncogene-like addiction to aneuploidy in human cancers.
  JOURNAL   Science. 2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521.
  PUBMED    37410869
REFERENCE   2  (bases 1 to 11759)
  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: "Science. 
            2023 Jul 6:eadg4521. doi: 10.1126/science.adg4521."
COMMENT     Sequence Label: UCK2_Deletion_gRNA_1
FEATURES             Location/Qualifiers
     source          1..11759
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     437..456
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(675..779)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(805..1260)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(942..963)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1154..1173
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1387..1409
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1401..1418
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1402..1418
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1428..1447
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1428..1446
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      1467..1602
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1642..1763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1653..1672)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1707..1726
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             1841..2074
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    2146..2734
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2252..2263
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     2661..2681
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(2750..3463,3464..3466,3467..3469))
                     /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             complement(join(2753..3463,3464..3466,3467..3469))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2795..2816)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3144..3163
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(3470..3526)
                     /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             complement(3542..3589)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(3590..7690)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(3590..7690)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7715..7735)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7742..7765)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      7765..7774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7794..8005)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    8074..8191
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        8198..8446
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8198..8218
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8369..8388
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    9040..9273
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    9766..9891
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             9938..10118
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(10119..10345)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(10373..10393)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10373..10391)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10412..10428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10412..10428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10425..10447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10436..10452
                     /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(10460..10466,10467..10484,10485..10490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10505..10526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10643..10660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10814..11402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10894..10913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11573..11759,1..605,606..674))
                     /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"
ORIGIN
        1 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
       61 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
      121 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
      181 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
      241 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
      301 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
      361 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
      421 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
      481 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
      541 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
      601 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
      661 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
      721 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
      781 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
      841 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
      901 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
      961 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     1021 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     1081 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     1141 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     1201 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     1261 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     1321 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     1381 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     1441 tatagggcga attgggtacg tccctaggcc tccaaaaaag cctcctcact acttctggaa
     1501 tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     1561 gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg
     1621 gatagctaga gccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     1681 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     1741 ttataagctg caataaacaa gttcctctca ctctctgata ttcatttctt tgcaagttat
     1801 aaatactgaa taataagatg acatgaacta ctatacgtac tgctagagat tttccacact
     1861 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
     1921 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
     1981 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaagc aaaaagcaga
     2041 tcttgtcttc gttgggagtg aattagccct tccagtcccc ccttttcttt taaaaagtgg
     2101 ctaagatcta cagctgcctt gtaagtcatt ggtcttaaag tcgacgcggg gaggcggccc
     2161 aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg
     2221 gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg tagcagaagg
     2281 acgtcccgcg cagaatccag gtggcaacac aggcgagcag ccaaggaaag gacgatgatt
     2341 tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg
     2401 caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg aaagtcccgg
     2461 aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag
     2521 tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga gacagcaacc
     2581 aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag
     2641 gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca
     2701 atctttcaca aattttgtaa tccagaggtt gattgtcgac ttaacgcgtc tacttgtaca
     2761 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     2821 cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg
     2881 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     2941 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     3001 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     3061 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     3121 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     3181 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     3241 cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg
     3301 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     3361 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     3421 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg gggccggggt
     3481 tctcctccac gtcgccggcc tgcttcagca ggctgaagtt ggtggcgccg ctgccgctag
     3541 ctttcttttt cttagcttga ccagctttct tagtagcagc aggacgcttg tcgcctccca
     3601 gctgagacag gtcgatccgt gtctcgtaca ggccggtgat gctctggtgg atcagggtgg
     3661 cgtccagcac ctctttggtg ctggtgtacc tcttccggtc gatggtggtg tcaaagtact
     3721 tgaaggcggc aggggctccc agattggtca gggtaaacag gtggatgata ttctcggcct
     3781 gctctctgat gggcttatcc cggtgcttgt tgtaggcgga cagcactttg tccagattag
     3841 cgtcggccag gatcactctc ttggagaact cgctgatctg ctcgatgatc tcgtccaggt
     3901 agtgcttgtg ctgttccaca aacagctgtt tctgctcatt atcctcgggg gagcccttca
     3961 gcttctcata gtggctggcc aggtacagga agttcacata tttggagggc agggccagtt
     4021 cgtttccctt ctgcagttcg ccggcagagg ccagcattct cttccggccg ttttccagct
     4081 cgaacaggga gtacttaggc agcttgatga tcaggtcctt tttcacttct ttgtagccct
     4141 tggcttccag aaagtcgatg ggattcttct cgaagctgct tctttccatg atggtgatcc
     4201 ccagcagctc tttcacactc ttcagtttct tggacttgcc cttttccact ttggccacca
     4261 ccagcacaga ataggccacg gtggggctgt cgaagccgcc gtacttctta gggtcccagt
     4321 ccttctttct ggcgatcagc ttatcgctgt tcctcttggg caggatagac tctttgctga
     4381 agccgcctgt ctgcacctcg gtctttttca cgatattcac ttggggcatg ctcagcactt
     4441 tccgcacggt ggcaaaatcc cggcccttat cccacacgat ctccccggtt tcgccgtttg
     4501 tctcgatcag aggccgcttc cggatctcgc cgttggccag ggtaatctcg gtcttgaaaa
     4561 agttcatgat gttgctgtag aagaagtact tggcggtagc cttgccgatt tcctgctcgc
     4621 tcttggcgat catcttccgc acgtcgtaca ccttgtagtc gccgtacacg aactcgcttt
     4681 ccagcttagg gtactttttg atcagggcgg ttcccacgac ggcgttcagg taggcgtcgt
     4741 gggcgtggtg gtagttgttg atctcgcgca ctttgtaaaa ctggaaatcc ttccggaaat
     4801 cggacaccag cttggacttc agggtgatca ctttcacttc ccggatcagc ttgtcattct
     4861 cgtcgtactt agtgttcatc cgggagtcca ggatctgtgc cacgtgcttt gtgatctgcc
     4921 gggtttccac cagctgtctc ttgatgaagc cggccttatc cagttcgctc aggccgcctc
     4981 tctcggcctt ggtcagattg tcgaactttc tctgggtaat cagcttggcg ttcagcagct
     5041 gccgccagta gttcttcatc ttcttcacga cctcttcgga gggcacgttg tcgctcttgc
     5101 cccggttctt gtcgcttctg gtcagcacct tgttgtcgat ggagtcgtcc ttcagaaagc
     5161 tctgaggcac gatatggtcc acatcgtagt cggacagccg gttgatgtcc agttcctggt
     5221 ccacgtacat atcccgccca ttctgcaggt agtacaggta cagcttctcg ttctgcagct
     5281 gggtgttttc cacggggtgt tctttcagga tctggctgcc cagctctttg atgccctctt
     5341 cgatccgctt cattctctcg cggctgttct tctgtccctt ctgggtggtc tggttctctc
     5401 tggccatttc gatcacgatg ttctcgggct tgtgccggcc catcactttc acgagctcgt
     5461 ccaccacctt cactgtctgc aggatgccct tcttaatggc ggggctgccg gccagattgg
     5521 caatgtgctc gtgcaggcta tcgccctggc cggacacctg ggctttctgg atgtcctctt
     5581 taaaggtcag gctgtcgtcg tggatcagct gcatgaagtt tctgttggcg aagccgtcgg
     5641 acttcaggaa atccaggatt gtcttgccgg actgcttgtc ccggatgccg ttgatcagct
     5701 tccggctcag cctgccccag ccggtgtatc tccgccgctt cagctgcttc atcactttgt
     5761 cgtcgaacag gtgggcatag gttttcagcc gttcctcgat catctctctg tcctcaaaca
     5821 gtgtcagggt cagcacgata tcttccagaa tgtcctcgtt ttcctcattg tccaggaagt
     5881 ccttgtcctt gataattttc agcagatcgt ggtatgtgcc cagggaggcg ttgaaccgat
     5941 cttccacgcc ggagatttcc acggagtcga agcactcgat tttcttgaag tagtcctctt
     6001 tcagctgctt cacggtcact ttccggttgg tcttgaacag caggtccacg atggcctttt
     6061 tctgctcgcc gctcaggaag gcgggctttc tcattccctc ggtcacgtat ttcactttgg
     6121 tcagctcgtt atacacggtg aagtactcgt acagcaggct gtgcttgggc agcaccttct
     6181 cgttgggcag gttcttatcg aagttggtca tccgctcgat gaagctctgg gcggaagcgc
     6241 ccttgtccac cacttcctcg aagttccagg gggtgatggt ttcctcgctc tttctggtca
     6301 tccaggcgaa tctgctgttt cccctggcca gagggcccac gtagtagggg atgcggaagg
     6361 tcaggatctt ctcgatcttt tcccggttgt ccttcaggaa tgggtaaaaa tcttcctgcc
     6421 gccgcagaat ggcgtgcagc tctcccaggt ggatctggtg ggggatgctg ccgttgtcga
     6481 aggtccgctg cttccgcagc aggtcctctc tgttcagctt cacgagcagt tcctcggtgc
     6541 cgtccatctt ttccaggatg ggcttgatga acttgtagaa ctcttcctgg ctggctccgc
     6601 cgtcaatgta gccggcgtag ccgttcttgc tctggtcgaa gaaaatctct ttgtacttct
     6661 caggcagctg ctgccgcacg agagctttca gcagggtcag gtcctggtgg tgctcgtcgt
     6721 atctcttgat catagaggcg ctcagggggg ccttggtgat ctcggtgttc actctcagga
     6781 tgtcgctcag caggatggcg tcggacaggt tcttggcggc cagaaacagg tcggcgtact
     6841 ggtcgccgat ctgggccagc aggttgtcca ggtcgtcgtc gtaggtgtcc ttgctcagct
     6901 gcagtttggc atcctcggcc aggtcgaagt tgctcttgaa gttgggggtc aggcccaggc
     6961 tcagggcaat caggtttccg aacaggccat tcttcttctc gccgggcagc tgggcgatca
     7021 gattttccag ccgtctgctc ttgctcagtc tggcagacag gatggccttg gcgtccacgc
     7081 cgctggcgtt gatggggttt tcctcgaaca gctggttgta ggtctgcacc agctggatga
     7141 acagcttgtc cacgtcgctg ttgtcggggt tcaggtcgcc ctcgatcagg aagtggcccc
     7201 ggaacttgat catgtgggcc agggccagat agatcagccg caggtcggcc ttgtcggtgc
     7261 tgtccaccag tttctttctc aggtggtaga tggtggggta cttctcgtgg taggccacct
     7321 cgtccacgat gttgccgaag atggggtgcc gctcgtgctt cttatcctct tccaccagga
     7381 aggactcttc cagtctgtgg aagaagctgt cgtccacctt ggccatctcg ttgctgaaga
     7441 tctcttgcag atagcagatc cggttcttcc gtctggtgta tcttcttctg gcggttctct
     7501 tcagccgggt ggcctcggct gtttcgccgc tgtcgaacag cagggctccg atcaggttct
     7561 tcttgatgct gtgccggtcg gtgttgccca gcaccttgaa tttcttgctg ggcaccttgt
     7621 actcgtcggt gatcacggcc cagcccacag agttggtgcc gatgtccagg ccgatgctgt
     7681 acttcttgtc ggctgctggg actccgtgga taccgacctt ccgcttcttc tttggggcca
     7741 tcttatcgtc atcgtctttg taatccatgg tggcggatcc cgcgtcacga cacctgtgtt
     7801 ctggcggcaa acccgttgcg aaaaagaacg ttcacggcga ctactgcact tatatacggt
     7861 tctcccccac cctcgggaaa aaggcggagc cagtacacga catcactttc ccagtttacc
     7921 ccgcgccacc ttctctaggc accggttcaa ttgccgaccc ctccccccaa cttctcgggg
     7981 actgtgggcg atgtgcgctc tgcccactga cgggcaccgg agccactcga gccggcgcca
     8041 aagtggatct ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt
     8101 ttgtaattct ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact
     8161 gtacccccca atcccccctt ttcttttaaa attgtgggag ggcctatttc ccatgattcc
     8221 ttcatatttg catatacgat acaaggctgt tagagagata attagaatta atttgactgt
     8281 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
     8341 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
     8401 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgatg aagttcaggt
     8461 tgtacgagtt tcagagctat gcacaagagt gcatagcaag ttgaaataag gctagtccgt
     8521 tatcaacttt gctggaaaca gcaaagtggc accgagtcgg tgcttttttt gcgtgaccga
     8581 gcttgtctgc ccgcgcagtc cactagacag acgcgcagag gtcgggcggg cctagggcca
     8641 atttcccatg gttcccttca tttgcatata tgatgtaata aggttatgga gactattaaa
     8701 cttagcccta atcaaactat atgatgataa tgtgtgtggt acaaaaggtc ataacttatt
     8761 atatactttg aaacttaaaa aagggttaca gtttagtcac cataactgta aaattttttc
     8821 tattcttagc tttatatagt tcttgagagg ccatgtttat cgttagaagt tagcgagccc
     8881 aggtttaaga gctatgctgg aaacagcata gcaagtttaa ataaggctag tccgttatca
     8941 actttgctgg aaacagcaaa gtggcaccga gtcggtgctt tttttgaatt ctagatcttg
     9001 agacaaatgg cgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
     9061 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
     9121 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
     9181 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
     9241 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
     9301 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
     9361 tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc
     9421 agcggccgct tgctgtgcgg tggtcttact tttgttttgc tcttcctcta tcttgtctaa
     9481 agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg ttgctactgt
     9541 attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta gctgtcccag
     9601 tatttgtcta cagccttctg atgtttctaa caggccagga ttaactgcga atcgttctag
     9661 ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc ctggccttaa
     9721 ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc tctcgcaccc
     9781 atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca gtcgccgccc
     9841 ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg
     9901 tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt ccacactgac
     9961 taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc acacactact
    10021 tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag ttagccagag
    10081 agctcccagg ctcagatctg gtctaaccag agagacccgt ttatgtatcg agctaggcac
    10141 ttaaatacaa tatctctgca atgcggcaat tcagtggttc gtccaatcca tgtcagaccc
    10201 gtctgttgcc ttcctaataa ggcacgatcg taccacctta cttccaccaa tcggcatgca
    10261 cggtgctttt tctctccttg taaggcatgt tgctaactca tcgttaccat gttgcaagac
    10321 tacaagagta ttgcataaga ctacattaag cttgcagctc cagcttttgt tccctttagt
    10381 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
    10441 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
    10501 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
    10561 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
    10621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    10681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    10741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    10801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    10861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    10921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    10981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    11041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    11101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    11161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    11221 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    11281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    11341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    11401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    11461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    11521 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    11581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    11641 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    11701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagcca
//
