LOCUS       sequence-420116-       11946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CasRx (NLS-RfxCas13d-NLS) with 2A-Blasticidin for RNA targeting. 
            CasRx-2A-Blasticidin is driven by EF1a promoter. EGFP is driven by 
            SV40.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11946)
  AUTHORS   Wang L, Bitar M, Lu X, Jacquelin S, Nair S, Sivakumaran H, Hillman 
            KM, Kaufmann S, Ziegman R, Casciello F, Gowda H, Rosenbluh J, 
            Edwards SL, French JD
  TITLE     CRISPR-Cas13d screens identify KILR, a breast cancer risk-associated
            lncRNA that regulates DNA replication and repair.
  JOURNAL   Mol Cancer. 2024 May 15;23(1):101. doi: 10.1186/s12943-024-02021-y.
  PUBMED    38745269
REFERENCE   2  (bases 1 to 11946)
  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 
            Cancer."; date: "2024-05-15"; volume: "23(1):101. doi"; pages: " 
            10.1186/s12943-024-02021-y"
COMMENT     Sequence Label: pLX_TRC311/NLS-Cas13d-P2A-Blast
FEATURES             Location/Qualifiers
     source          1..11946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     290..307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    424..445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(460..465,466..483,484..490)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    498..514
                     /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     503..525
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     522..538
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     522..538
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     557..577
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        559..577
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        605..831
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1059..1184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1677..1910
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2095..2139
                     /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"
     promoter        2370..2580
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2392..2588
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2439..2574
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(2607..2609,2610..2612,2613..3326)
                     /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(2607..2609,2610..2612,2613..3323)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2652..2673)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2913..2932)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3260..3281
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3367..3484
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3622..4800
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3853..4791
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4748..4768
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4818..4838
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4848..7745
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKNGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA
                     HINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             7755..7775
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7785..7811
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7821..7877
                     /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             7878..8276
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(8279..8303)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             8311..8352
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    8403..8991
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8456..8476)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8874..8885)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9063..9296
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9374..9495
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9411..9430)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9465..9484
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9535..9670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9690..9709)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9691..9709)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9719..9736)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9719..9735)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9728..9750)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9877..10332
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9964..9983)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10174..10195
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10358..10462
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10463..10531,10532..11323)
                     /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(10681..10700)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11494..11946,1..136)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
       61 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      421 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      481 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
      541 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
      601 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
      661 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      721 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      781 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1081 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1141 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1201 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1261 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1321 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1381 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1441 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1501 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     1561 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1621 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1681 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1741 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1801 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1861 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1921 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1981 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     2041 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     2101 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     2161 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     2221 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     2281 tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat tagtcagcaa
     2341 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcgtctca
     2401 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2461 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2521 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2581 tttgcaaaaa gctttctaga ggtaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     2641 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2701 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2761 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2821 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2881 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2941 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3001 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3061 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     3121 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     3181 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     3241 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     3301 tcggcatgga cgagctgtac aagtagaatt ctagatcttg agacaaatgg cagtattcat
     3361 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     3421 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     3481 ttttcgggtt tattacaggg acagcagaga tccactttgg cgccggctcg aggcctgcag
     3541 gtgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag
     3601 gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg cacatcgccc
     3661 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg
     3721 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     3781 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     3841 gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat
     3901 ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct tgatcccgag
     3961 cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc cccttcgcct
     4021 cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac
     4081 cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt ttgatgacct
     4141 gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga tctgcacact
     4201 ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt
     4261 tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta gtctcaagct
     4321 ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca
     4381 aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc cggccctgct
     4441 gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga gtcacccaca
     4501 caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg
     4561 gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc tttaggttgg
     4621 ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac tgaagttagg
     4681 ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt tggatcttgg
     4741 ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca ggtgtcgtga
     4801 ggctagccac catgagcccc aagaagaaga gaaaggtgga ggccagcatc gaaaaaaaaa
     4861 agtccttcgc caagggcatg ggcgtgaagt ccacactcgt gtccggctcc aaagtgtaca
     4921 tgacaacctt cgccgaaggc agcgacgcca ggctggaaaa gatcgtggag ggcgacagca
     4981 tcaggagcgt gaatgagggc gaggccttca gcgctgaaat ggccgataaa aacgccggct
     5041 ataagatcgg caacgccaaa ttcagccatc ctaagggcta cgccgtggtg gctaacaacc
     5101 ctctgtatac aggacccgtc cagcaggata tgctcggcct gaaggaaact ctggaaaaga
     5161 ggtacttcgg cgagagcgct gatggcaatg acaatatttg tatccaggtg atccataaca
     5221 tcctggacat tgaaaaaatc ctcgccgaat acattaccaa cgccgcctac gccgtcaaca
     5281 atatctccgg cctggataag gacattattg gattcggcaa gttctccaca gtgtatacct
     5341 acgacgaatt caaagacccc gagcaccata gggccgcttt caacaataac gataagctca
     5401 tcaacgccat caaggcccag tatgacgagt tcgacaactt cctcgataac cccagactcg
     5461 gctatttcgg ccaggccttt ttcagcaagg agggcagaaa ttacatcatc aattacggca
     5521 acgaatgcta tgacattctg gccctcctga gcggactgag gcactgggtg gtccataaca
     5581 acgaagaaga gtccaggatc tccaggacct ggctctacaa cctcgataag aacctcgaca
     5641 acgaatacat ctccaccctc aactacctct acgacaggat caccaatgag ctgaccaact
     5701 ccttctccaa gaactccgcc gccaacgtga actatattgc cgaaactctg ggaatcaacc
     5761 ctgccgaatt cgccgaacaa tatttcagat tcagcattat gaaagagcag aaaaacctcg
     5821 gattcaatat caccaagctc agggaagtga tgctggacag gaaggatatg tccgagatca
     5881 ggaaaaatca taaggtgttc gactccatca ggaccaaggt ctacaccatg atggactttg
     5941 tgatttatag gtattacatc gaagaggatg ccaaggtggc tgccgccaat aagtccctcc
     6001 ccgataatga gaagtccctg agcgagaagg atatctttgt gattaacctg aggggctcct
     6061 tcaacgacga ccagaaggat gccctctact acgatgaagc taatagaatt tggagaaagc
     6121 tcgaaaatat catgcacaac atcaaggaat ttaggggaaa caagacaaga gagtataaga
     6181 agaaggacgc ccctagactg cccagaatcc tgcccgctgg ccgtgatgtt tccgccttca
     6241 gcaaactcat gtatgccctg accatgttcc tggatggcaa ggagatcaac gacctcctga
     6301 ccaccctgat taataaattc gataacatcc agagcttcct gaaggtgatg cctctcatcg
     6361 gagtcaacgc taagttcgtg gaggaatacg cctttttcaa agactccgcc aagatcgccg
     6421 atgagctgag gctgatcaag tccttcgcta gaatgggaga acctattgcc gatgccagga
     6481 gggccatgta tatcgacgcc atccgtattt taggaaccaa cctgtcctat gatgagctca
     6541 aggccctcgc cgacaccttt tccctggacg agaacggaaa caagctcaag aatggcaagc
     6601 acggcatgag aaatttcatt attaataacg tgatcagcaa taaaaggttc cactacctga
     6661 tcagatacgg tgatcctgcc cacctccatg agatcgccaa aaacgaggcc gtggtgaagt
     6721 tcgtgctcgg caggatcgct gacatccaga aaaaacaggg ccagaacggc aagaaccaga
     6781 tcgacaggta ctacgaaact tgtatcggaa aggataaggg caagagcgtg agcgaaaagg
     6841 tggacgctct cacaaagatc atcaccggaa tgaactacga ccaattcgac aagaaaagga
     6901 gcgtcattga ggacaccggc agggaaaacg ccgagaggga gaagtttaaa aagatcatca
     6961 gcctgtacct caccgtgatc taccacatcc tcaagaatat tgtcaatatc aacgccaggt
     7021 acgtcatcgg attccattgc gtcgagcgtg atgctcaact gtacaaggag aaaggctacg
     7081 acatcaatct caagaaactg gaagagaagg gattcagctc cgtcaccaag ctctgcgctg
     7141 gcattgatga aactgccccc gataagagaa aggacgtgga aaaggagatg gctgaaagag
     7201 ccaaggagag cattgacagc ctcgagagcg ccaaccccaa gctgtatgcc aattacatca
     7261 aatacagcga cgagaagaaa gccgaggagt tcaccaggca gattaacagg gagaaggcca
     7321 aaaccgccct gaacgcctac ctgaggaaca ccaagtggaa tgtgatcatc agggaggacc
     7381 tcctgagaat tgacaacaag acatgtaccc tgttcagaaa caaggccgtc cacctggaag
     7441 tggccaggta tgtccacgcc catatcaacg acattgccga ggtcaattcc tacttccaac
     7501 tgtaccatta catcatgcag agaattatca tgaatgagag gtacgagaaa agcagcggaa
     7561 aggtgtccga gtacttcgac gctgtgaatg acgagaagaa gtacaacgat aggctcctga
     7621 aactgctgtg tgtgcctttc ggctactgta tccccaggtt taagaacctg agcatcgagg
     7681 ccctgttcga taggaacgag gccgccaagt tcgacaagga gaaaaagaag gtgtccggca
     7741 attccggatc cggacctaag aaaaagagga aggtggcggc cgcttaccca tatgatgtcc
     7801 ccgactatgc cggatccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg
     7861 aagagaatcc tggaccgatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     7921 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     7981 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     8041 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     8101 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     8161 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     8221 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaaaccc
     8281 ccaactttct tgtacaaagt ggttgatatc ggtaagccta tccctaaccc tctcctcggt
     8341 ctcgattcta cgtagtaatg aactagtacc ggttaagtcg acaatcaacg cgttaagtcg
     8401 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     8461 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     8521 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     8581 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     8641 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     8701 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     8761 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     8821 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     8881 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     8941 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt
     9001 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     9061 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     9121 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     9181 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     9241 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtac
     9301 gtatagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     9361 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     9421 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     9481 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg
     9541 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     9601 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     9661 tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     9721 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     9781 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     9841 cttcccaaca gttgcgcagc ctgaatggcg aatcggacgc gccctgtagc ggcgcattaa
     9901 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     9961 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
    10021 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
    10081 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
    10141 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
    10201 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
    10261 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
    10321 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
    10381 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
    10441 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
    10501 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
    10561 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
    10621 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
    10681 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
    10741 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
    10801 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
    10861 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
    10921 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
    10981 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
    11041 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
    11101 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
    11161 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
    11221 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
    11281 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
    11341 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
    11401 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
    11461 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
    11521 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
    11581 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
    11641 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
    11701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
    11761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
    11821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
    11881 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
    11941 cggcag
//
