LOCUS       sequence-252606-       10303 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral expression of ESR1 E380Q.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10303)
  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 10303)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-ESR1-E380Q
FEATURES             Location/Qualifiers
     source          1..10303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2223..3401
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2454..3392
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3349..3369
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    3421..3445
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4297..5232
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLQCAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             4345..5238
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLQCAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     protein_bind    complement(5240..5264)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5317..5358
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    5379..5946
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5396..5948
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5540..5557)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5767..5786
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5960..5962,5963..5965,5966..6679)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5960..5962,5963..5965,5966..6676)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6005..6026)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6266..6285)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6613..6634
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    6693..7281
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6746..6766)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7164..7175)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7356..7589
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7708..7726)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7794..7898
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7899..7967,7968..8759)
                     /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(8117..8136)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8930..9518
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9419..9438
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9672..9689
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9929..9950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9965..9970,9971..9988,9989..9995)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10003..10019
                     /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     10008..10030
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10027..10043
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10027..10043
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10093..10112
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtcgt
     2221 gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     2281 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2341 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2401 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2461 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2521 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2581 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2641 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2701 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2761 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     2821 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     2881 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     2941 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     3001 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     3061 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3121 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3181 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3241 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3301 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3361 cagtggttca aagttttttt cttccatttc aggtgtcgtg aagcggccct gcagatatca
     3421 acaagtttgt acaaaaaagc aggcttcacc atgaccatga ccctccacac caaagcatct
     3481 gggatggccc tactgcatca gatccaaggg aacgagctgg agcccctgaa ccgtccgcag
     3541 ctcaagatcc ccctggagcg gcccctgggc gaggtgtacc tggacagcag caagcccgcc
     3601 gtgtacaact accccgaggg cgccgcctac gagttcaacg ccgcggccgc cgccaacgcg
     3661 caggtctacg gtcagaccgg cctcccctac ggccccgggt ctgaggctgc ggcgttcggc
     3721 tccaacggcc tggggggttt ccccccactc aacagcgtgt ctccgagccc gctgatgcta
     3781 ctgcacccgc cgccgcagct gtcgcctttc ctgcagcccc acggccagca ggtgccctac
     3841 tacctggaga acgagcccag cggctacacg gtgcgcgagg ccggcccgcc ggcattctac
     3901 aggccaaatt cagataatcg acgccagggt ggcagagaaa gattggccag taccaatgac
     3961 aagggaagta tggctatgga atctgccaag gagactcgct actgtgcagt gtgcaatgac
     4021 tatgcttcag gctaccatta tggagtctgg tcctgtgagg gctgcaaggc cttcttcaag
     4081 agaagtattc aaggacataa cgactatatg tgtccagcca ccaaccagtg caccattgat
     4141 aaaaacagga ggaagagctg ccaggcctgc cggctccgca aatgctacga agtgggaatg
     4201 atgaaaggtg ggatacgaaa agaccgaaga ggagggagaa tgttgaaaca caagcgccag
     4261 agagatgatg gggagggcag gggtgaagtg gggtctgctg gagacatgag agctgccaac
     4321 ctttggccaa gcccgctcat gatcaaacgc tctaagaaga acagcctggc cttgtccctg
     4381 acggccgacc agatggtcag tgccttgttg gatgctgagc cccccatact ctattccgag
     4441 tatgatccta ccagaccctt cagtgaagct tcgatgatgg gcttactgac caacctggca
     4501 gacagggagc tggttcacat gatcaactgg gcgaagaggg tgccaggctt tgtggatttg
     4561 accctccatg atcaggtcca ccttctacaa tgtgcctggc tagagatcct gatgattggt
     4621 ctcgtctggc gctccatgga gcacccaggg aagctactgt ttgctcctaa cttgctcttg
     4681 gacaggaacc agggaaaatg tgtagagggc atggtggaga tcttcgacat gctgctggct
     4741 acatcatctc ggttccgcat gatgaatctg cagggagagg agtttgtgtg cctcaaatct
     4801 attattttgc ttaattctgg agtgtacaca tttctgtcca gcaccctgaa gtctctggaa
     4861 gagaaggacc atatccaccg agtcctggac aagatcacag acactttgat ccacctgatg
     4921 gccaaggcag gcctgaccct gcagcagcag caccagcggc tggcccagct cctcctcatc
     4981 ctctcccaca tcaggcacat gagtaacaaa ggcatggagc atctgtacag catgaagtgc
     5041 aagaacgtgg tgcccctcta tgacctgctg ctggagatgc tggacgccca ccgcctacat
     5101 gcgcccacta gccgtggagg ggcatccgtg gaggagacgg accaaagcca cttggccact
     5161 gcgggctcta cttcatcgca ttccttgcaa aagtattaca tcacggggga ggcagagggt
     5221 ttccctgcca cggtctgaga cccagctttc ttgtacaaag tggttgatat ccagcacagt
     5281 ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
     5341 ctcggtctcg attctacgcg taccggttag taatgagatc cctccccccc ccctaacgtt
     5401 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     5461 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     5521 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     5581 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     5641 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     5701 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     5761 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     5821 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     5881 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     5941 tgataatatg gccacacata tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     6001 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     6061 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     6121 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     6181 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     6241 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     6301 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     6361 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     6421 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     6481 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     6541 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     6601 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     6661 ggacgagctg tacaagtaaa tcgatagatc ctaatcaacc tctggattac aaaatttgtg
     6721 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     6781 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     6841 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     6901 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     6961 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     7021 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     7081 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     7141 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     7201 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     7261 ccctttgggc cgcctccccg cctgagatcc tttaagacca atgacttaca aggcagctgt
     7321 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     7381 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     7441 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     7501 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     7561 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     7621 tttataactt gcaaagaaat gaatatcaga gagtgagagg cccgggttaa ttaaggaaag
     7681 ggctagatca ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     7741 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     7801 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     7861 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     7921 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     7981 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     8041 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     8101 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     8161 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     8221 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     8281 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     8341 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     8401 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     8461 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     8521 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     8581 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     8641 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     8701 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     8761 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     8821 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     8881 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     8941 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     9001 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     9061 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     9121 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     9181 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     9241 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     9301 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     9361 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     9421 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     9481 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     9541 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     9601 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     9661 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     9721 ctctccccgc gcgttggccg attcattaat gcagcaagct catggctgac taattttttt
     9781 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     9841 cttttttgga ggcctaggct tttgcaaaaa gctccccgtg gcacgacagg tttcccgact
     9901 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     9961 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
    10021 ttcacacagg aaacagctat gacatgatta cgaatttcac aaataaagca tttttttcac
    10081 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
    10141 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
    10201 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
    10261 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
