LOCUS       sequence-394148-        9940 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses kinase-inactive VRK2 (K168E).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9940)
  AUTHORS   So J, Mabe NW, Englinger B, Chow KH, Moyer SM, Yerrum S, Trissal MC,
            Marques JG, Kwon JJ, Shim B, Pal S, Panditharatna E, Quinn T, 
            Schaefer DA, Jeong D, Mayhew DL, Hwang J, Beroukhim R, Ligon KL, 
            Stegmaier K, Filbin MG, Hahn WC
  TITLE     VRK1 as a synthetic lethal target in VRK2 promoter-methylated 
            cancers of the nervous system.
  JOURNAL   JCI Insight. 2022 Oct 10;7(19):e158755. doi: 
            10.1172/jci.insight.158755.
  PUBMED    36040810
REFERENCE   2  (bases 1 to 9940)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "JCI 
            Insight."; date: "2022-10-10"; volume: "7(19):e158755. doi"; pages: 
            " 10.1172/jci.insight.158755"
COMMENT     Sequence Label: pLX317-VRK2-K168E
FEATURES             Location/Qualifiers
     source          1..9940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        84..1262
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          315..1253
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1210..1230
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1282..1306
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             2446..2457
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    complement(2837..2861)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2869..2910
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2961..3549
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3014..3034)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3432..3443)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3621..3854
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3932..4053
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3969..3988)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4023..4042
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4093..4228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4248..4267)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4249..4267)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4277..4294)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4277..4293)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4286..4308)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4435..4890
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4522..4541)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4732..4753
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4916..5020
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5021..5089,5090..5881)
                     /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(5239..5258)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6052..6640
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6541..6560
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6794..6811
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6928..6949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6964..6969,6970..6987,6988..6994)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7002..7018
                     /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     7007..7029
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7026..7042
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7026..7042
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7061..7081
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7063..7081
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7109..7335
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7336..7516
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7563..7688
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8181..8414
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8599..8643
                     /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        8874..9084
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        8896..9092
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8943..9078
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             9124..9726
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(9124..9143)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9620..9640
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9769..9886
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 aggtgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct
       61 aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc
      121 ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt
      181 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      241 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      301 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
      361 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
      421 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
      481 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
      541 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
      601 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
      661 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
      721 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
      781 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
      841 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
      901 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
      961 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     1021 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     1081 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     1141 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     1201 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     1261 gaggctagca tcgattgatc aacaagtttg tacaaaaaag caggcaccat gccaccaaaa
     1321 agaaatgaaa aatacaaact tcctattcca tttccagaag gcaaggttct ggatgatatg
     1381 gaaggcaatc agtgggtact gggcaagaag attggctctg gaggatttgg attgatatat
     1441 ttagctttcc ccacaaataa accagagaaa gatgcaagac atgtagtaaa agtggaatat
     1501 caagaaaatg gcccgttatt ttcagaactt aaattttatc agagagttgc aaaaaaagac
     1561 tgtatcaaaa agtggataga acgcaaacaa cttgattatt taggaattcc tctgttttat
     1621 ggatctggtc tgactgaatt caagggaaga agttacagat ttatggtaat ggaaagacta
     1681 ggaatagatt tacagaagat ctcaggccag aatggtacct ttaaaaagtc aactgtcctg
     1741 caattaggta tccgaatgtt ggatgtactg gaatatatac atgaaaatga atatgttcat
     1801 ggtgatatag aagcagcaaa tctacttttg ggttacaaaa atccagacca ggtttatctt
     1861 gcagattatg gactttccta cagatattgt cccaatggga accacaaaca gtatcaggaa
     1921 aatcctagaa aaggccataa tgggacaata gagtttacca gcttggatgc ccacaaggga
     1981 gtagccttgt ccagacgaag tgacgttgag atcctcggct actgcatgct gcggtggttg
     2041 tgtgggaaac ttccctggga acagaacctg aaggaccctg tggctgtgca gactgctaaa
     2101 acaaatctgt tggacgagct cccccagtca gtgcttaaat gggctccttc tggaagcagt
     2161 tgctgtgaaa tagcccaatt tttggtatgt gctcatagtt tagcatatga tgaaaagcca
     2221 aactatcaag ccctcaagaa aattttgaac cctcatggaa tacctttagg accactggac
     2281 ttttccacaa aaggacagag tataaatgtc catactccaa acagtcaaaa agttgattca
     2341 caaaaggctg caacaaagca agtcaacaag gcacacaata ggttaatcga aaaaaaagtc
     2401 cacagtgaga gaagcgctga gtcctgtgca acatggaaag tgcagaaaga ggagaaactg
     2461 attggattga tgaacaatga agcagctcag gaaagcacaa ggagaagaca gaaatatcaa
     2521 gagtctcaag aacctttgaa tgaagtaaac agtttcccac aaaaaatcag ctatacacaa
     2581 ttcccaaact cattttatga gcctcatcaa gattttacca gtccagatat attcaagaag
     2641 tcaagatctc catcttggta taaatacact tccacagtca gcacggggat cacagactta
     2701 gaaagttcaa ctggactttg gcctacaatt tcccagttta ctcttagtga agagacaaac
     2761 gcagatgttt attattatcg catcatcata cctgtccttt tgatgttagt atttcttgct
     2821 ttattttttc tctgagaccc agctttcttg tacaaagtgg ttgatatcgg taagcctatc
     2881 cctaaccctc tcctcggtct cgattctacg tagtaatgaa ctagtaccgg ttaagtcgac
     2941 aatcaacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     3001 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3061 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3121 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3181 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3241 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3301 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     3361 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3421 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3481 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3541 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     3601 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     3661 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     3721 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     3781 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     3841 gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt atttataact
     3901 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     3961 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4021 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     4081 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4141 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4201 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     4261 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     4321 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     4381 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     4441 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4501 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4561 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4621 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4681 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4741 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4801 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4861 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4921 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4981 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5041 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5101 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5161 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5221 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5281 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5341 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5401 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5461 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5521 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5581 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5641 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5701 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5761 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5821 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5881 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5941 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6001 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6061 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6121 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     6181 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     6241 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     6301 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     6361 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6421 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6481 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6541 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6601 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6661 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6721 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6781 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6841 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6901 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6961 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     7021 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     7081 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
     7141 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
     7201 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
     7261 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
     7321 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     7381 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     7441 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     7501 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     7561 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     7621 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     7681 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     7741 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     7801 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     7861 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     7921 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     7981 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     8041 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     8101 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     8161 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     8221 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     8281 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     8341 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     8401 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     8461 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     8521 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     8581 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     8641 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     8701 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     8761 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccatg
     8821 cattgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     8881 gaagtatgca aagcatgcgt ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     8941 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     9001 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     9061 gaggcttttt tggaggccta ggcttttgca aaaagctttc tagaggtacc accggagctt
     9121 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     9181 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     9241 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     9301 gacatcggca aggtgtgggt cgcggacgac ggcgccgccg tggcggtctg gaccacgccg
     9361 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     9421 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     9481 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     9541 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     9601 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     9661 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     9721 agttaacgaa ttctagatct tgagacaaat ggcagtattc atccacaatt ttaaaagaaa
     9781 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     9841 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     9901 ggacagcaga gatccacttt ggcgccggct cgaggcctgc
//
