LOCUS       sequence-252209-       12593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of IGF1R E1356K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12593)
  AUTHORS   Ng PK, Li J, Jeong KJ, Shao S, Chen H, Tsang YH, Sengupta S, Wang Z,
            Bhavana VH, Tran R, Soewito S, Minussi DC, Moreno D, Kong K, 
            Dogruluk T, Lu H, Gao J, Tokheim C, Zhou DC, Johnson AM, Zeng J, Ip 
            CKM, Ju Z, Wester M, Yu S, Li Y, Vellano CP, Schultz N, Karchin R, 
            Ding L, Lu Y, Cheung LWT, Chen K, Shaw KR, Meric-Bernstam F, Scott 
            KL, Yi S, Sahni N, Liang H, Mills GB
  TITLE     Systematic Functional Annotation of Somatic Mutations in Cancer.
  JOURNAL   Cancer Cell. 2018 Mar 12;33(3):450-462.e10. doi: 
            10.1016/j.ccell.2018.01.021.
  PUBMED    29533785
REFERENCE   2  (bases 1 to 12593)
  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"; doi: 
            "10.1016/j.ccell.2018.01"; journalName: "Cancer Cell"; date: 
            "2018-03-12- 12"; volume: "33"; issue: "3"; pages: "450-462.e10"
COMMENT     Sequence Label: pHAGE-IGF1R-E1356K
FEATURES             Location/Qualifiers
     source          1..12593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1199..1223
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5331..5355)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5363..5381
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5406..5425)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5407..5425)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    complement(5431..5451)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             5505..5546
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5567..6134
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5584..6136
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5728..5745)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5955..5974
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6148..6747
                     /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(6148..6167)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6644..6664
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6761..7349
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6814..6834)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7232..7243)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7424..7657
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7776..7794)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7862..7966
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7967..8035,8036..8827)
                     /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(8185..8204)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8998..9586
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9487..9506
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9740..9757
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9997..10018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10033..10038,10039..10056,10057..10063)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10071..10087
                     /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     10076..10098
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10095..10111
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10095..10111
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10161..10180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             10372..11005
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    11052..11177
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11674..11907
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12092..12136
                     /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    12429..12546
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgaa gcggccctgc agatatcaac
     1201 aagtttgtac aaaaaagcag gcaccatgaa gtctggctcc ggaggagggt ccccgacctc
     1261 gctgtggggg ctcctgtttc tctccgccgc gctctcgctc tggccgacga gtggagaaat
     1321 ctgcgggcca ggcatcgaca tccgcaacga ctatcagcag ctgaagcgcc tggagaactg
     1381 cacggtgatc gagggctacc tccacatcct gctcatctcc aaggccgagg actaccgcag
     1441 ctaccgcttc cccaagctca cggtcattac cgagtacttg ctgctgttcc gagtggctgg
     1501 cctcgagagc ctcggagacc tcttccccaa cctcacggtc atccgcggct ggaaactctt
     1561 ctacaactac gccctggtca tcttcgagat gaccaatctc aaggatattg ggctttacaa
     1621 cctgaggaac attactcggg gggccatcag gattgagaaa aatgctgacc tctgttacct
     1681 ctccactgtg gactggtccc tgatcctgga tgcggtgtcc aataactaca ttgtggggaa
     1741 taagccccca aaggaatgtg gggacctgtg tccagggacc atggaggaga agccgatgtg
     1801 tgagaagacc accatcaaca atgagtacaa ctaccgctgc tggaccacaa accgctgcca
     1861 gaaaatgtgc ccaagcacgt gtgggaagcg ggcgtgcacc gagaacaatg agtgctgcca
     1921 ccccgagtgc ctgggcagct gcagcgcgcc tgacaacgac acggcctgtg tagcttgccg
     1981 ccactactac tatgccggtg tctgtgtgcc tgcctgcccg cccaacacct acaggtttga
     2041 gggctggcgc tgtgtggacc gtgacttctg cgccaacatc ctcagcgccg agagcagcga
     2101 ctccgagggg tttgtgatcc acgacggcga gtgcatgcag gagtgcccct cgggcttcat
     2161 ccgcaacggc agccagagca tgtactgcat cccttgtgaa ggtccttgcc cgaaggtctg
     2221 tgaggaagaa aagaaaacaa agaccattga ttctgttact tctgctcaga tgctccaagg
     2281 atgcaccatc ttcaagggca atttgctcat taacatccga cgggggaata acattgcttc
     2341 agagctggag aacttcatgg ggctcatcga ggtggtgacg ggctacgtga agatccgcca
     2401 ttctcatgcc ttggtctcct tgtccttcct aaaaaacctt cgcctcatcc taggagagga
     2461 gcagctagaa gggaattact ccttctacgt cctcgacaac cagaacttgc agcaactgtg
     2521 ggactgggac caccgcaacc tgaccatcaa agcagggaaa atgtactttg ctttcaatcc
     2581 caaattatgt gtttccgaaa tttaccgcat ggaggaagtg acggggacta aagggcgcca
     2641 aagcaaaggg gacataaaca ccaggaacaa cggggagaga gcctcctgtg aaagtgacgt
     2701 cctgcatttc acctccacca ccacgtcgaa gaatcgcatc atcataacct ggcaccggta
     2761 ccggccccct gactacaggg atctcatcag cttcaccgtt tactacaagg aagcaccctt
     2821 taagaatgtc acagagtatg atgggcagga tgcctgcggc tccaacagct ggaacatggt
     2881 ggacgtggac ctcccgccca acaaggacgt ggagcccggc atcttactac atgggctgaa
     2941 gccctggact cagtacgccg tttacgtcaa ggctgtgacc ctcaccatgg tggagaacga
     3001 ccatatccgt ggggccaaga gtgagatctt gtacattcgc accaatgctt cagttccttc
     3061 cattcccttg gacgttcttt cagcatcgaa ctcctcttct cagttaatcg tgaagtggaa
     3121 ccctccctct ctgcccaacg gcaacctgag ttactacatt gtgcgctggc agcggcagcc
     3181 tcaggacggc tacctttacc ggcacaatta ctgctccaaa gacaaaatcc ccatcaggaa
     3241 gtatgccgac ggcaccatcg acattgagga ggtcacagag aaccccaaga ctgaggtgtg
     3301 tggtggggag aaagggcctt gctgcgcctg ccccaaaact gaagccgaga agcaggccga
     3361 gaaggaggag gctgaatacc gcaaagtctt tgagaatttc ctgcacaact ccatcttcgt
     3421 gcccagacct gaaaggaagc ggagagatgt catgcaagtg gccaacacca ccatgtccag
     3481 ccgaagcagg aacaccacgg ccgcagacac ctacaacatc accgacccgg aagagctgga
     3541 gacagagtac cctttctttg agagcagagt ggataacaag gagagaactg tcatttctaa
     3601 ccttcggcct ttcacattgt accgcatcga tatccacagc tgcaaccacg aggctgagaa
     3661 gctgggctgc agcgcctcca acttcgtctt tgcaaggact atgcccgcag aaggagcaga
     3721 tgacattcct gggccagtga cctgggagcc aaggcctgaa aactccatct ttttaaagtg
     3781 gccggaacct gagaatccca atggattgat tctaatgtat gaaataaaat acggatcaca
     3841 agttgaggat cagcgagaat gtgtgtccag acaggaatac aggaagtatg gaggggccaa
     3901 gctaaaccgg ctaaacccgg ggaactacac agcccggatt caggccacat ctctctctgg
     3961 gaatgggtcg tggacagatc ctgtgttctt ctatgtccag gccaaaacag gatatgaaaa
     4021 cttcatccat ctgatcatcg ctctgcccgt cgctgtcctg ttgatcgtgg gagggttggt
     4081 gattatgctg tacgtcttcc atagaaagag aaataacagc aggctgggga atggagtgct
     4141 gtatgcctct gtgaacccgg agtacttcag cgctgctgat gtgtacgttc ctgatgagtg
     4201 ggaggtggct cgggagaaga tcaccatgag ccgggaactt gggcaggggt cgtttgggat
     4261 ggtctatgaa ggagttgcca agggtgtggt gaaagatgaa cctgaaacca gagtggccat
     4321 taaaacagtg aacgaggccg caagcatgcg tgagaggatt gagtttctca acgaagcttc
     4381 tgtgatgaag gagttcaatt gtcaccatgt ggtgcgattg ctgggtgtgg tgtcccaagg
     4441 ccagccaaca ctggtcatca tggaactgat gacacggggc gatctcaaaa gttatctccg
     4501 gtctctgagg ccagaaatgg agaataatcc agtcctagca cctccaagcc tgagcaagat
     4561 gattcagatg gccggagaga ttgcagacgg catggcatac ctcaacgcca ataagttcgt
     4621 ccacagagac cttgctgccc ggaattgcat ggtagccgaa gatttcacag tcaaaatcgg
     4681 agattttggt atgacgcgag atatctatga gacagactat taccggaaag gagggaaagg
     4741 gctgctgccc gtgcgctgga tgtctcctga gtccctcaag gatggagtct tcaccactta
     4801 ctcggacgtc tggtccttcg gggtcgtcct ctgggagatc gccacactgg ccgagcagcc
     4861 ctaccagggc ttgtccaacg agcaagtcct tcgcttcgtc atggagggcg gccttctgga
     4921 caagccagac aactgtcctg acatgctgtt tgaactgatg cgcatgtgct ggcagtataa
     4981 ccccaagatg aggccttcct tcctggagat catcagcagc atcaaagagg agatggagcc
     5041 tggcttccgg gaggtctcct tctactacag cgaggagaac aagctgcccg agccggagga
     5101 gctggacctg gagccagaga acatggagag cgtccccctg gacccctcgg cctcctcgtc
     5161 ctccctgcca ctgcccgaca gacactcagg acacaaggcc gagaacggcc ccggccctgg
     5221 ggtgctggtc ctccgcgcca gcttcgacga gagacagcct tacgcccaca tgaacggggg
     5281 ccgcaagaac aagcgggcct tgccgctgcc ccagtcttcg acctgctaga acccagcttt
     5341 cttgtacaaa gtggtgatat ccaattaacc ctcactaaag gtagttaaag tatagttgtt
     5401 tgagtcccta tagtgagtcg tattaccacc caacttttct atacaaagtg gttgatatcc
     5461 agcacagtgg cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta
     5521 accctctcct cggtctcgat tctacgcgta ccggttagta atgagatccc tccccccccc
     5581 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     5641 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     5701 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     5761 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     5821 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     5881 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     5941 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     6001 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     6061 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     6121 aaacacgatg ataatatggc cacacatatg accgagtaca agcccacggt gcgcctcgcc
     6181 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     6241 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     6301 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     6361 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     6421 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     6481 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     6541 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     6601 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     6661 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     6721 tgcatgaccc gcaagcccgg tgcctaaatc gatagatcct aatcaacctc tggattacaa
     6781 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     6841 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     6901 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     6961 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     7021 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     7081 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     7141 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     7201 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     7261 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     7321 tcggatctcc ctttgggccg cctccccgcc tgagatcctt taagaccaat gacttacaag
     7381 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     7441 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     7501 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7561 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7621 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     7681 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc cgggttaatt
     7741 aaggaaaggg ctagatcatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     7801 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     7861 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     7921 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7981 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     8041 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     8101 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     8161 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     8221 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     8281 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     8341 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     8401 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     8461 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     8521 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     8581 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     8641 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     8701 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     8761 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     8821 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8881 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8941 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     9001 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     9061 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     9121 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     9181 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     9241 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     9301 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     9361 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     9421 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     9481 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     9541 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     9601 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     9661 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     9721 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     9781 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agcaagctca tggctgacta
     9841 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     9901 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccgtggc acgacaggtt
     9961 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
    10021 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
    10081 ataacaattt cacacaggaa acagctatga catgattacg aatttcacaa ataaagcatt
    10141 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    10201 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
    10261 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
    10321 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag ttggaagggc
    10381 taattcactc ccaaagaaga caagatatcc ttgatctgtg gatctaccac acacaaggct
    10441 acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca ctgacctttg
    10501 gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc aataaaggag
    10561 agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag
    10621 tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga gagctgcatc
    10681 cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc gctggggact
    10741 ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga tcctgcatat
    10801 aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg agcctgggag
    10861 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
    10921 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
    10981 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
    11041 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
    11101 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
    11161 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
    11221 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
    11281 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
    11341 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
    11401 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
    11461 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
    11521 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
    11581 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
    11641 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
    11701 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
    11761 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
    11821 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
    11881 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
    11941 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
    12001 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
    12061 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
    12121 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
    12181 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
    12241 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
    12301 gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag
    12361 atccattcga ttagtgaacg gatctcgacg gtatcgccga attcacaaat ggcagtattc
    12421 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
    12481 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
    12541 aattttcggg tttattacag ggacagcaga gatccagttt ggactagtcg tga
//
