LOCUS       sequence-360487-       10235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express EML4-ALK fusion variant 3a (E6a;A20) harboring ALK I1171N 
            mutation in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10235)
  AUTHORS   Shiba-Ishii A, Johnson TW, Dagogo-Jack I, Mino-Kenudson M, Johnson 
            TR, Wei P, Weinrich SL, McTigue MA, Walcott MA, Nguyen-Phuong L, 
            Dionne K, Acker A, Kiedrowski LA, Do A, Peterson JL, Barth JL, Yeap 
            BY, Gainor JF, Lin JJ, Yoda S, Hata AN
  TITLE     Analysis of lorlatinib analogs reveals a roadmap for targeting 
            diverse compound resistance mutations in ALK-positive lung cancer.
  JOURNAL   Nat Cancer. 2022 Jun 20. pii: 10.1038/s43018-022-00399-6. doi: 
            10.1038/s43018-022-00399-6.
  PUBMED    35726063
REFERENCE   2  (bases 1 to 10235)
  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: "Nat Cancer.
            2022 Jun 20. pii: 10.1038/s43018-022-00399-6. doi: 
            10.1038/s43018-022-00399-6."
COMMENT     Sequence Label: pLenti-EML4-ALK variant 3a I1171N
FEATURES             Location/Qualifiers
     source          1..10235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     79..99
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     125..149
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     224..243
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        224..242
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    272..296
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2691..2715)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2747..2770
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2786..2812
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             2786..2812
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2819..3385
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2835..3387
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2979..2996)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3206..3225
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3399..3998
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3399..3418)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3895..3915
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4015..4603
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4068..4088)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4486..4497)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4678..4911
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5030..5048)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5116..5220
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5221..5289,5290..6081)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5439..5458)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6252..6840
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6741..6760
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6994..7011
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7251..7272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7287..7292,7293..7310,7311..7317)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7325..7341
                     /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     7330..7352
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7349..7365
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7349..7365
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7415..7434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             7626..8259
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    8306..8431
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8928..9161
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9346..9390
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    9687..9804
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        9856..10159
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(10160..10235,1..128)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
       61 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      121 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
      181 tccatagaag acaccggcgg ccgcagatct ctcgaggttg atctaatacg actcactata
      241 gggagaccca agctggctag ttaagctatc aacaagtttg tacaaaaaag caggctccgc
      301 ggccgccccc ttcaccatgg acggtttcgc cggcagtctc gatgatagta tttctgctgc
      361 aagtacttct gatgttcaag atcgcctgtc agctcttgag tcacgagttc agcaacaaga
      421 agatgaaatc actgtgctaa aggcggcttt ggctgatgtt ttgaggcgtc ttgcaatctc
      481 tgaagatcat gtggcctcag tgaaaaaatc agtctcaagt aaaggccaac caagccctcg
      541 agcagttatt cccatgtcct gtataaccaa tggaagtggt gcaaacagaa aaccaagtca
      601 taccagtgct gtctcaattg caggaaaaga aactctttca tctgctgcta aaagtggtac
      661 agaaaaaaag aaagaaaaac cacaaggaca gagagaaaaa aaagaggaat ctcattctaa
      721 tgatcaaagt ccacaaattc gagcatcacc ttctccccag ccctcttcac aacctctcca
      781 aatacacaga caaactccag aaagcaagaa tgctactccc accaaaagca taaaacgacc
      841 atcaccagct gaaaagtcac ataattcttg ggaaaattca gatgatagcc gtaataaatt
      901 gtcgaaaata ccttcaacac ccaaattaat accaaaagtt accaaaactg cagacaagca
      961 taaagatgtc atcatcaacc aagtgtaccg ccggaagcac caggagctgc aagccatgca
     1021 gatggagctg cagagccctg agtacaagct gagcaagctc cgcacctcga ccatcatgac
     1081 cgactacaac cccaactact gctttgctgg caagacctcc tccatcagtg acctgaagga
     1141 ggtgccgcgg aaaaacatca ccctcattcg gggtctgggc catggcgcct ttggggaggt
     1201 gtatgaaggc caggtgtccg gaatgcccaa cgacccaagc cccctgcaag tggctgtgaa
     1261 gacgctgcct gaagtgtgct ctgaacagga cgaactggat ttcctcatgg aagccctgat
     1321 caacagcaaa ttcaaccacc agaacattgt tcgctgcatt ggggtgagcc tgcaatccct
     1381 gccccggttc atcctgctgg agctcatggc ggggggagac ctcaagtcct tcctccgaga
     1441 gacccgccct cgcccgagcc agccctcctc cctggccatg ctggaccttc tgcacgtggc
     1501 tcgggacatt gcctgtggct gtcagtattt ggaggaaaac cacttcatcc accgagacat
     1561 tgctgccaga aactgcctct tgacctgtcc aggccctgga agagtggcca agattggaga
     1621 cttcgggatg gcccgagaca tctacagggc gagctactat agaaagggag gctgtgccat
     1681 gctgccagtt aagtggatgc ccccagaggc cttcatggaa ggaatattca cttctaaaac
     1741 agacacatgg tcctttggag tgctgctatg ggaaatcttt tctcttggat atatgccata
     1801 ccccagcaaa agcaaccagg aagttctgga gtttgtcacc agtggaggcc ggatggaccc
     1861 acccaagaac tgccctgggc ctgtataccg gataatgact cagtgctggc aacatcagcc
     1921 tgaagacagg cccaactttg ccatcatttt ggagaggatt gaatactgca cccaggaccc
     1981 ggatgtaatc aacaccgctt tgccgataga atatggtcca cttgtggaag aggaagagaa
     2041 agtgcctgtg aggcccaagg accctgaggg ggttcctcct ctcctggtct ctcaacaggc
     2101 aaaacgggag gaggagcgca gcccagctgc cccaccacct ctgcctacca cctcctctgg
     2161 caaggctgca aagaaaccca cagctgcaga ggtctctgtt cgagtcccta gagggccggc
     2221 cgtggaaggg ggacacgtga atatggcatt ctctcagtcc aaccctcctt cggagttgca
     2281 caaggtccac ggatccagaa acaagcccac cagcttgtgg aacccaacgt acggctcctg
     2341 gtttacagag aaacccacca aaaagaataa tcctatagca aagaaggagc cacacgacag
     2401 gggtaacctg gggctggagg gaagctgtac tgtcccacct aacgttgcaa ctgggagact
     2461 tccgggggcc tcactgctcc tagagccctc ttcgctgact gccaatatga aggaggtacc
     2521 tctgttcagg ctacgtcact tcccttgtgg gaatgtcaat tacggctacc agcaacaggg
     2581 cttgccctta gaagccgcta ctgcccctgg agctggtcat tacgaggata ccattctgaa
     2641 aagcaagaat agcatgaacc agcctgggcc ctgaaagggt gggcgcgccg acccagcttt
     2701 cttgtacaaa gtggtttagc tagagggccc gcggttcgaa ctcgaggact acaaggacga
     2761 cgatgacaag tcggccgctg gaggataccc ctacgacgtg cccgactacg cctagaattc
     2821 cgcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     2881 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     2941 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     3001 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     3061 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     3121 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     3181 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     3241 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     3301 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     3361 tttcctttga aaaacacgat gataatatgg ccacaaccat gaccgagtac aagcccacgg
     3421 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     3481 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     3541 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     3601 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     3661 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     3721 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     3781 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     3841 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     3901 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     3961 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatg catcgatgga tcctaatcaa
     4021 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     4081 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     4141 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     4201 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     4261 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     4321 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     4381 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     4441 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     4501 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     4561 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctgagat cctttaagac
     4621 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     4681 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     4741 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4801 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     4861 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     4921 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     4981 ggcccgggtt aattaaggaa agggctagat cattcttgaa gacgaaaggg cctcgtgata
     5041 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5101 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5161 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5221 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5281 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5341 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5401 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5461 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5521 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5581 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5641 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5701 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5761 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5821 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5881 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5941 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6001 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6061 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6121 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6181 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6241 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6301 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6361 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6421 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6481 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6541 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6601 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6661 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6721 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6781 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6841 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6901 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6961 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7021 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagcaag
     7081 ctcatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     7141 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctccccg
     7201 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7261 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7321 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaatttc
     7381 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     7441 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
     7501 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
     7561 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
     7621 gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta
     7681 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata
     7741 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga
     7801 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga
     7861 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc
     7921 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact
     7981 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct
     8041 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga
     8101 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     8161 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     8221 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     8281 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     8341 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     8401 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     8461 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     8521 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     8581 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     8641 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     8701 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     8761 acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg
     8821 agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc
     8881 aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc
     8941 cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta
     9001 caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt
     9061 gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga
     9121 atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct
     9181 ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg
     9241 gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca
     9301 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
     9361 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
     9421 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
     9481 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
     9541 cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag
     9601 agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg ccgaattaat
     9661 tcacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     9721 gcagggaaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     9781 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     9841 actagtggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     9901 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     9961 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
    10021 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
    10081 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
    10141 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
    10201 gtttgactca cggggatttc caagtctcca cccca
//
