LOCUS       sequence-310699-       14925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the NINJA targeting construct (inducible neoantigen 
            expression via cumulative Cre, rtTA, doxycycline and tamoxifen)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14925)
  AUTHORS   Damo M, Fitzgerald B, Lu Y, Nader M, William I, Cheung JF, Connolly 
            KA, Foster GG, Akama-Garren E, Lee DY, Chang GP, Gocheva V, Schmidt 
            LM, Boileve A, Wilson JH, Cui C, Monroy I, Gokare P, Cabeceiras P, 
            Jacks T, Joshi NS
  TITLE     Inducible de novo expression of neoantigens in tumor cells and mice.
  JOURNAL   Nat Biotechnol. 2020 Jul 27. pii: 10.1038/s41587-020-0613-1. doi: 
            10.1038/s41587-020-0613-1.
  PUBMED    32719479
REFERENCE   2  (bases 1 to 14925)
  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 
            Biotechnol. 2020 Jul 27. pii: 10.1038/s41587-020-0613-1. doi: 
            10.1038/s41587-020-0613-1."
COMMENT     Sequence Label: NINJA TC
FEATURES             Location/Qualifiers
     source          1..14925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        74..178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(179..247,248..1039)
                     /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(397..416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1210..1798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1699..1718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1952..1969
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2122..2127,2128..2145,2146..2152)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2160..2176
                     /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     2165..2187
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2184..2200
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2184..2200
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2219..2239
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2221..2239
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     3093..3109
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3094..3110
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3143..3176)
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(3263..3862)
                     /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(3346..3366)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3843..3862
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3883..4382)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3931..3950)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     4324..4347
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    complement(4406..4439)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    complement(4406..4439)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             5046..5981
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             5094..5987
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     protein_bind    6070..6103
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    6118..6151
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    6118..6151
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             complement(6996..7016)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(7050..7364)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(7053..7077)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    7124..7142
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7160..7178
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7195..7213
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7231..7249
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7267..7285
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7302..7320
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7338..7356
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     enhancer        9847..10226
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10228..10504
                     /label=chicken beta-actin promoter
     intron          10505..11521
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     11445..11467
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     11529..11548
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(11573..11575,11576..11578,11579..12037)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMA"
     primer_bind     complement(11618..11639)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(11879..11898)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    12160..12193
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    12244..12291
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(12333..12356)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(12457..12490)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     polyA_signal    12511..12592
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     12536..12555
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(12701..12748)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     13119..13140
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(13213..13231)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(13214..13231)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(13251..13268)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13257..13481
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(14331..14350)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(14332..14350)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(14360..14377)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(14360..14376)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14369..14391)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(14518..14925,1..48)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14605..14624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     14815..14836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
       61 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      121 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      181 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      241 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      301 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      361 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      421 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      481 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      541 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
      601 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
      661 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
      721 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
      781 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
      841 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
      901 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
      961 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1021 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1081 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1141 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1201 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1261 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1321 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1381 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1441 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1501 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1561 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     1621 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     1681 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1741 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     1801 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     1861 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     1921 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     1981 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     2041 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     2101 cagtcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     2161 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc
     2221 aattaaccct cactaaaggg aacaaaagct ggagctctgt tttggttggc gtaaggcgcc
     2281 tgtcagttaa cggcagccgg agtgcgcagc cgccggcagc ctcgctctgc ccactgggtg
     2341 gggcgggagg taggtggggt gaggcgagct ggacgtgcgg gcgcggtcgg cctctggcgg
     2401 ggcgggggrg gggagggagg gtcagcgaaa gtagctcgcg cgcgagcggc cgcccaccct
     2461 ccccttcctc tgggggagtc gttttacccg ccgccggccg ggcctcgtcg tctgattggc
     2521 tctcggggcc cagaaaactg gcccttgcca ttggctcgtg ttcgtgcaag ttgagtccat
     2581 ccgccggcca gcgggggcgg cgaggaggcg ctcccaggtt ccggccctcc cctcggcccc
     2641 gcgccgcaga gtctggccgc gcgcccctgc gcaacgtggc aggaagcgcg cgctgggggc
     2701 ggggacgggc agtagggctg agcggctgcg gggcgggtgc aagcacgttt ccgacttgag
     2761 ttgcctcaag aggggcgtgc tgagccagac ctccatcgcg cactccgggg agtggaggga
     2821 aggagcgagg gctcagttgg gctgttttgg aggcaggaag cacttgctct cccaaagtcg
     2881 ctctgagttg ttatcagtaa gggagctgca gtggagtagg cggggagaag gccgcaccct
     2941 tctccggagg ggggagggga gtgttgcaat acctttctgg gagttctctg ctgcctcctg
     3001 gcttctgagg accgccctgg gcctgggaga atcccttccc cctcttccct cgtgatctgc
     3061 aactccagtc tttctcccgg tgctagctac tctcgaggtc gacggtatcg ataagcttga
     3121 tatcgaattc ctgcagcccg ggataacttc gtatagtaca cattatacga agttatggct
     3181 aagctttgtg ccatgccatt gatgctagga acaaacgacc caacacccgt gcgttttatt
     3241 ctgtcttttt attgccacgc gttcaggcac cgggcttgcg ggtcatgcac caggtgcgcg
     3301 gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga
     3361 ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca
     3421 ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga
     3481 cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct
     3541 gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga
     3601 acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt
     3661 ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct
     3721 cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg
     3781 cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca
     3841 ccgtgggctt gtactcggtc atggtaagct tgggctgcag gtcgaaaggc ccggagatga
     3901 ggaagaggag aacagcgcgg cagacgtgcg cttttgaagc gtgcagaatg ccgggcctcc
     3961 ggaggacctt cgggcgcccg ccccgcccct gagcccgccc ctgagcccgc ccccggaccc
     4021 accccttccc agcctctgag cccagaaagc gaaggagcaa agctgctatt ggccgctgcc
     4081 ccaaaggcct acccgcttcc attgctcagc ggtgctgtcc atctgcacga gactagtgag
     4141 acgtgctact tccatttgtc acgtcctgca cgacgcgagc tgcggggcgg gggggaactt
     4201 cctgactagg ggaggagtag aaggtggcgc gaaggggcca ccaaagaacg gagccggttg
     4261 gcgcctaccg gtggatgtgg aatgtgtgcg aggccagagg ccacttgtgt agcgccaagt
     4321 gcccagcggg gctgctaaag cgcatgctcc agactgcctt gggaaaagcg cctcccctac
     4381 ccggtaccta gcacagcctg aatggataac ttcgtatagg ataccttata cgaagttata
     4441 ccggtttcaa aaaatggaag gaaatcagga actaactctc tgctcttgtt ttccaggaca
     4501 acctggtgcg cagctacaac aaggccctga agaagaacgc cccctacccc atcttcgcta
     4561 tcaagaacgg ccctaagagc cacatcggca ggcacctgat gaccagcttt ctgagcatga
     4621 agggcctgac cgagctgaca aacgtggtgg gcaactggag cgacaagagg gcctccgccg
     4681 tggccaggac cacctacacc caccagatca ccgccatccc cgaccactac ttcgccctgg
     4741 tgtccaggta ctacgcctac gaccccatca gcaaggagat gatcgccctg aaggacgaga
     4801 ccaaccccat cgaggagtgg cagcacatcg agcagctgaa gggcagcgcc gagggcagca
     4861 tcagataccc cgcctggaac ggcatcatca gccaggaggt gctggactac ctgagcagct
     4921 acatcaacag gcggatttcc gtacgcggat ccaaaggtgg gatacgaaaa gaccgaagag
     4981 gagggagaat gttgaaacac aagcgccaga gagatgatgg ggagggcagg ggtgaagtgg
     5041 ggtctgctgg agacatgaga gctgccaacc tttggccaag cccgctcatg atcaaacgct
     5101 ctaagaagaa cagcctggcc ttgtccctga cggccgacca gatggtcagt gccttgttgg
     5161 atgctgagcc ccccatactc tattccgagt atgatcctac cagacccttc agtgaagctt
     5221 cgatgatggg cttactgacc aacctggcag acagggagct ggttcacatg atcaactggg
     5281 cgaagagggt gccaggcttt gtggatttga ccctccatga tcaggtccac cttcttgaat
     5341 gtgcctggct agagatcctg atgattggtc tcgtctggcg ctccatggag cacccagtga
     5401 agctactgtt tgctcctaac ttgctcttgg acaggaacca gggaaaatgt gtagagggca
     5461 tggtggagat cttcgacatg ctgctggcta catcatctcg gttccgcatg atgaatctgc
     5521 agggagagga gtttgtgtgc ctcaaatcta ttattttgct taattctgga gtgtacacat
     5581 ttctgtccag caccctgaag tctctggaag agaaggacca tatccaccga gtcctggaca
     5641 agatcacaga cactttgatc cacctgatgg ccaaggcagg cctgaccctg cagcagcagc
     5701 accagcggct ggcccagctc ctcctcatcc tctcccacat caggcacatg agtaacaaag
     5761 gcatggagca tctgtacagc atgaagtgca agaacgtggt gcccctctat gacctgctgc
     5821 tggaggcggc ggacgcccac cgcctacatg cgcccactag ccgtggaggg gcatccgtgg
     5881 aggagacgga ccaaagccac ttggccactg cgggctctac ttcatcgcat tccttgcaaa
     5941 agtattacat cacgggggag gcagagggtt tccctgccac agcttgaacg cgtggcaata
     6001 aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cggcgcgcca gtaacatgta
     6061 gtagtaaaca taacttcgta taatgtgtac tatacgaagt tatggcttct agctgagata
     6121 acttcgtata aggtatccta tacgaagtta tggatctatt aactcacctt cagcagctgg
     6181 tactcctgct tgttgctgct gctgttgccg gtcctgttca ctctcttcag cacgggctcg
     6241 ctgttcctca ggaactcgtc caggtacacc agggggtcga tcctgcctct ggcgctgaaa
     6301 aagtagatgt gcctggacac gcttgtcttg gtctcggtca ccaggcactg aatgatcacg
     6361 cccaggtact tgttctgcac cagcttgaag ctcttggggt ccacgttctt gatgtcgctg
     6421 aacctgccgc agttgatgaa tgtggccagg aacaggaact ggtacagggt cttggtcttg
     6481 gtgaacctgc tggtgtactc gaagctgttc aggatcttct cggtgatctc ccagatgctc
     6541 tcgccctcgg acagcagggc cttcagcatc ttcttgctgt ggctgttgcc cttgtcggcc
     6601 tcctcgctgc tctcgaactg cagctgcagg ctggacacga tgtcggtgat gtcgctctgg
     6661 tgcttctggc cgttgtaagg gatgatggtg aactcccagg cggggatcag cttcttcagg
     6721 ctggcctcca ggatggtggc cttctgggtc ttgtacttga actgcaggct cttgttcacg
     6781 atgtcgaagc tcaggctgtt gctgatgatg gtgttgtagc tcatgaaggt ggccctcttg
     6841 atggcggtgc cgttgtgggt gatcatccag cacaggtagg tcagctcggc ggcacagctg
     6901 gcgatcttct cgccgctggg cctctcgaat ctctccacga actgccgcac cagcaccttg
     6961 gggggggtct tgcacaggat gtcgaactgg ctcatcacct tcctcttctt cttaggagcc
     7021 atggtgtcga gtgacttacg tattctccag gcgatctgac ggttcactaa acgagctctg
     7081 cttatatagg cctcccaccg tacacgccta cctcgacata cgttctctat cactgatagg
     7141 gagtaaactc gacatacgtt ctctatcact gatagggata aactcgacat acgttctcta
     7201 tcactgatag ggagtaaact cgacatacgt tctctatcac tgatagggag taaactcgac
     7261 atacgttctc tatcactgat agggagtaaa ctcgacatcg ttctctatca ctgataggga
     7321 gtaaactcga catacgttct ctatcactga tagggagtaa actcgacata tcgaaaccgt
     7381 cgcggccgcc agtgtgatgg atgactctag agggacagcc cccccccaaa gcccccaggg
     7441 atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg
     7501 gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg
     7561 aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca
     7621 cctgggggga tacggggaaa aagctttagg ctgaaagaga gatttagaat gacagaatca
     7681 tagaacggcc tgggttgcaa aggagcacag tgctcatcca gatccaaccc cctgctatgt
     7741 gcagggtcat caaccagcag cccaggctgc ccagagccac atccagcctg gccttgaatg
     7801 cctgcaggga tggggcatcc acagcctcct tgggcaacct gttcagtgcg tcaccaccct
     7861 ctgggggaaa aactgcctcc tcatatccaa cccaaacctc ccctgtctca gtgtaaagcc
     7921 attccccctt gtcctatcaa gggggagttt gctgtgacat tgttggtctg gggtgacaca
     7981 tgtttgccaa ttcagtgcat cacggagagg cagatcttgg ggataaggaa gtgcaggaca
     8041 gcatggacgt gggacatgca ggtgttgagg gctctgggac actctccaag tcacagcgtt
     8101 cagaacagcc ttaaggataa gaagatagga tagaaggaca aagagcaagt taaaacccag
     8161 catggagagg agcacaaaaa ggccacagac actgctggtc cctgtgtctg agcctgcatg
     8221 tttgatggtg tctggatgca agcagaaggg gtggaagagc ttgcctggag agatacagct
     8281 gggtcagtag gactgggaca ggcagctgga gaattgccat gtagatgttc atacaatcgt
     8341 caaatcatga aggctggaaa agccctccaa gatccccaag accaacccca acccacccac
     8401 cgtgcccact ggccatgtcc ctcagtgcca catccccaca gttcttcatc acctccaggg
     8461 acggtgaccc ccccacctcc gtgggcagct gtgccactgc agcaccgctc tttggagaag
     8521 gtaaatcttg ctaaatccag cccgaccctc ccctggcaca acgtaaggcc attatctctc
     8581 atccaactcc aggacggagt cagtgaggat ggggctctag agggacagcc cccccccaaa
     8641 gcccccaggg atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg
     8701 ctccgctccg gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc
     8761 gggcacgggg aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag
     8821 cctgcagaca cctgggggga tacggggaaa aagctttagg ctgaaagaga gatttagaat
     8881 gacagaatca tagaacggcc tgggttgcaa aggagcacag tgctcatcca gatccaaccc
     8941 cctgctatgt gcagggtcat caaccagcag cccaggctgc ccagagccac atccagcctg
     9001 gccttgaatg cctgcaggga tggggcatcc acagcctcct tgggcaacct gttcagtgcg
     9061 tcaccaccct ctgggggaaa aactgcctcc tcatatccaa cccaaacctc ccctgtctca
     9121 gtgtaaagcc attccccctt gtcctatcaa gggggagttt gctgtgacat tgttggtctg
     9181 gggtgacaca tgtttgccaa ttcagtgcat cacggagagg cagatcttgg ggataaggaa
     9241 gtgcaggaca gcatggacgt gggacatgca ggtgttgagg gctctgggac actctccaag
     9301 tcacagcgtt cagaacagcc ttaaggataa gaagatagga tagaaggaca aagagcaagt
     9361 taaaacccag catggagagg agcacaaaaa ggccacagac actgctggtc cctgtgtctg
     9421 agcctgcatg tttgatggtg tctggatgca agcagaaggg gtggaagagc ttgcctggag
     9481 agatacagct gggtcagtag gactgggaca ggcagctgga gaattgccat gtagatgttc
     9541 atacaatcgt caaatcatga aggctggaaa agccctccaa gatccccaag accaacccca
     9601 acccacccac cgtgcccact ggccatgtcc ctcagtgcca catccccaca gttcttcatc
     9661 acctccaggg acggtgaccc ccccacctcc gtgggcagct gtgccactgc agcaccgctc
     9721 tttggagaag gtaaatcttg ctaaatccag cccgaccctc ccctggcaca acgtaaggcc
     9781 attatctctc atccaactcc aggacggagt cagtgaggat ggggctctag aggatctagc
     9841 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     9901 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     9961 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
    10021 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
    10081 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
    10141 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
    10201 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
    10261 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
    10321 cagcgatggg ggcggggggg gggggggncg cgcgccrggs ggggcggggs ggggsgrggg
    10381 gsggggsggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
    10441 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
    10501 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
    10561 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
    10621 tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc tgtggctgcg
    10681 tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct cggggggtgc
    10741 gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga
    10801 gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc
    10861 cgggggcggt gccccgcggt gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg
    10921 tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct
    10981 gcacccccct ccccgagttg ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg
    11041 gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg
    11101 ggcggggccg cctcgggccg gggrgggctc gggggagggg cgcggcggcc cccggagcgc
    11161 cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt ttatggtaat cgtgcgagag
    11221 ggcgcaggga cttcctttgt cccaaatctg tgcggagccg aaatctggga ggcgccgccg
    11281 caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg
    11341 ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc cctctccagc ctcggggctg
    11401 tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg
    11461 tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca
    11521 gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaaa ccatggtgag
    11581 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
    11641 aaacggccac aagttcagcg tgtctggcga gggcgagggc gatgccacct acggcaagct
    11701 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
    11761 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
    11821 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
    11881 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
    11941 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
    12001 gtacaactac aacagccaca acgtctatat catggctagg tgcaaaatcc ccaacgacct
    12061 gaaacagaaa gttatgaacc acaaaggtga gtactaagta ccggagtcgc tgcgttgcct
    12121 tcgccccgtg ccccgctccg accgggaagt tcctattccg aagttcctat tcttcaaata
    12181 gtataggaac ttcgaattct gcagatatcc agcacagtgg cggccgctag catcgttacg
    12241 cgtgaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt cggatccgta
    12301 ctcaccttgt aaatgtcagg gccgttcaaa cccttgtcat cgtcgtcctt gtagtcgatc
    12361 ccgcatgttg caaagttgta cacagctggg gaaaaaaaag ggacaggata agtatgacat
    12421 catcaaggaa accctggact actgcgccct gggcccgaag ttcctatact atttgaagaa
    12481 taggaacttc ggaataggaa cttcaagctt gcagtgaaaa aaatgcttta tttgtgaaat
    12541 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
    12601 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
    12661 gtaaaacctc tacaaatgtg gtatggctga ttataagctt gaagttccta tactttctag
    12721 agaataggaa cttcggaata ggaacttcgg cgcgccgcag gacaactgca tgggggctgg
    12781 gatgggcata agaataaatg tctatgtgga cagccttaac cacttcagcc atgacctcta
    12841 tgtatgtttc taaccccaca gggagtttat cagtttaaat ccgttgagtt tgacgcagcc
    12901 gcagacctgt ctatctgtcg aagatccgcc gacaagcaga agaacggcat caaggcgaac
    12961 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
    13021 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
    13081 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
    13141 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtcga gcatgcatct
    13201 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
    13261 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
    13321 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
    13381 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
    13441 agacaatagc aggcatgctg gggatgcggt gggctctatg gctttatact acacctagtc
    13501 cgcggtgggc gggagtcttc tgggcaggct taaaggctaa cctggtgtgt gggcgttgtc
    13561 ctgcagggga attgaacagg tgtaaaattg gagggacaag acttcccaca gattttcggt
    13621 tttgtcggga agttttttaa taggggcaaa taaggaaaat gggaggatag gtagtcatct
    13681 ggggttttat gcagcaaaac tacaggttat tattgcttgt gatccgcctc ggagtatttt
    13741 ccatcgaggt agattaaaga catgctcacc cgagttttat actctcctgc ttgagatcct
    13801 tactacagta tgaaattaca gtgtcgcgag ttagactatg taagcagaat tttaatcatt
    13861 tttaaagagc ccagtacttc atatccattt ctcccgctcc ttctgcagcc ttatcaaaag
    13921 gtattttaga acactcattt tagccccatt ttcatttatt atactggctt atccaacccc
    13981 tagacagagc attggcattt tccctttcct gatcttagaa gtctgatgac tcatgaaacc
    14041 agacagatta gttacataca ccacaaatcg aggctgtagc tggggcctca acactgcagt
    14101 tcttttataa ctccttagta cactttttgt tgatcctttg ccttgatcct taattttcag
    14161 tgtctatcac ctctcccgtc aggtggtgtt ccacatttgg gcctattctc agtccaggga
    14221 gttttacaac aatagatgta ttgagaatcc aacctaaagc ttaactttcc actcccatga
    14281 atgcctctct cctttttctc catttataaa ctgagctggt acccaattcg ccctatagtg
    14341 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
    14401 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
    14461 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
    14521 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
    14581 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
    14641 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
    14701 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
    14761 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
    14821 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
    14881 ggattttgcc gatttcggcc tattggttaa aaaatgagct gattt
//
