LOCUS       sequence-369274-       11118 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses flag-tagged p53 with both R248W and P278A mutations in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11118)
  AUTHORS   Stransky LA, Vigeant SM, Huang B, West D, Denize T, Walton E, 
            Signoretti S, Kaelin WG Jr
  TITLE     Sensitivity of VHL mutant kidney cancers to HIF2 inhibitors does not
            require an intact p53 pathway.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Apr 5;119(14):e2120403119. doi: 
            10.1073/pnas.2120403119. Epub 2022 Mar 31.
  PUBMED    35357972
REFERENCE   2  (bases 1 to 11118)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2022-04-5"; volume: "119(14):e2120403119. 
            doi"; pages: " 10.1073/pnas.2120403119. Epub 2022 Mar 31"
COMMENT     Sequence Label: pLenti_EF1a_Flag_TP53_R248W+P278A
FEATURES             Location/Qualifiers
     source          1..11118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    14..38
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             44..67
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             83..1261
                     /codon_start=1
                     /gene="Homo sapiens TP53"
                     /product="human tumor suppressor protein that is often 
                     mutated in cancers"
                     /label=p53
                     /note="responds to cellular stresses to regulate the 
                     expression of target genes"
                     /translation="EEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLS
                     PDDIEQWFTEDPGPDEAPRMPEAAPRVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQG
                     SYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYK
                     QSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPP
                     EVGSDCTTIHYNYMCNSSCMGGMNWRPILTIITLEDSSGNLLGRNSFEVRVCACAGRDR
                     RTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMF
                     RELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
     protein_bind    complement(1263..1287)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1307..1330
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        1356..1855
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1391..1414)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1788..1807
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             1873..2898
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             1873..2898
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     protein_bind    complement(2929..2962)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3018..3606
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3071..3091)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3489..3500)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3609..3625)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3610..3626)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3815..4048
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4120..4241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4157..4176)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4211..4230
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4281..4416
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(4437..4455)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      complement(4478..4836)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4518..4539)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4730..4749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4863..4967
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4969..5326
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4969..5160
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4992..5012)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5177..5312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5361..6155
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5361..6155
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5415..5434)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6025..6044
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6343..6362)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6387..6434
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6763..7351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7252..7271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7526..7829
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7831..8029
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7984..8004
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             8047..8227
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8274..8399
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8896..9129
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9314..9358
                     /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    9656..9773
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        9845..11023
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          10076..11014
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     10971..10991
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        11040..11058
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gatctggcca atcacaagtt tgtacaaaaa agcaggctaa atggactaca aggacgacga
       61 tgacaagggt ggcggaggga gcgaggagcc gcagtcagat cctagcgtcg agccccctct
      121 gagtcaggaa acattttcag acctatggaa actacttcct gaaaacaacg ttctgtcccc
      181 cttgccgtcc caagcaatgg atgatttgat gctgtccccg gacgatatag agcagtggtt
      241 cactgaagac ccaggtccag atgaagctcc cagaatgcca gaggctgctc cccgcgtggc
      301 ccctgcacca gcagctccta caccggcggc ccctgcacca gccccctcct ggcccctgtc
      361 atcttctgtc ccttcccaga aaacctacca gggcagctac ggtttccgtc tgggcttctt
      421 gcattctggg acagccaagt ctgtgacttg cacgtactcc cctgccctca acaagatgtt
      481 ttgccaactg gccaagacct gccctgtgca gctgtgggtt gattccacac ccccgcccgg
      541 cacccgcgtc cgcgccatgg ccatctacaa gcagtcacag cacatgacgg aggttgtgag
      601 gcgctgcccc caccatgagc gctgctcaga tagcgatggt ctggcccctc ctcagcatct
      661 tatccgagtg gaaggaaatt tgcgtgtgga gtatttggat gacagaaaca cttttcgaca
      721 tagtgtggtg gtgccctatg agccgcctga ggttggctct gactgtacca ccatccacta
      781 caactacatg tgtaacagct cttgtatggg cggaatgaac tggaggccca tcctcaccat
      841 catcacactg gaagactcca gtggtaatct actgggacgg aacagctttg aggtgcgtgt
      901 ttgtgcctgt gctgggagag accggcgcac agaggaagag aatctccgca agaaagggga
      961 gcctcaccac gagctgcccc cagggagcac taagcgagca ctgcccaaca acaccagctc
     1021 ctctccccag ccaaagaaga aaccactgga tggagaatat ttcacccttc agatccgtgg
     1081 gcgtgagcgc ttcgagatgt tccgagagct gaatgaggcc ttggaactca aggatgccca
     1141 ggctgggaag gagccagggg ggagcagggc tcactccagc cacctgaagt ccaaaaaggg
     1201 tcagtctacc tcccgccata aaaaactcat gttcaagaca gaagggcctg actcagactg
     1261 aaacccagct ttcttgtaca aagtggtggg aggaggtgga tctggcgact acaaggacga
     1321 cgatgacaag tgactcgagg ttaacgaatt ctaccgggta ggggaggcgc ttttcccaag
     1381 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     1441 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     1501 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     1561 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     1621 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     1681 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     1741 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     1801 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     1861 catcccgcca ccatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     1921 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     1981 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     2041 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     2101 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     2161 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     2221 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     2281 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     2341 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     2401 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     2461 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     2521 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     2581 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     2641 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     2701 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     2761 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     2821 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     2881 ccgagggcaa aggaatagtg tacaagtagg aattcgtcga gggacctaat aacttcgtat
     2941 agcatacatt atacgaagtt atacatgttt aagggttccg gttccactag gtacaattcg
     3001 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3061 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3121 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3181 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3241 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3301 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3361 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3421 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     3481 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3541 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3601 ccccgcatcg ataccgtcga cctcgatcga gacctagaaa aacatggagc aatcacaagt
     3661 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     3721 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     3781 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     3841 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     3901 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     3961 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     4021 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     4081 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     4141 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4201 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     4261 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4321 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4381 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     4441 tagtgagtcg tattagcatg tgagcaaaag gccagcataa atcaaaagaa tagaccgaga
     4501 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4561 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4621 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4681 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4741 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4801 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4861 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4921 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4981 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5041 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5101 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5161 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5221 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     5281 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     5341 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5401 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5461 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5521 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5581 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5641 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5701 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5761 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5821 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5881 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5941 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6001 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6061 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6121 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6181 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6241 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6301 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     6361 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     6421 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6481 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6541 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6601 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6661 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6721 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6781 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6841 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6901 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6961 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7021 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7081 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7141 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7201 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7261 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7321 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7381 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7441 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
     7501 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     7561 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     7621 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     7681 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     7741 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     7801 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     7861 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     7921 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     7981 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc
     8041 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     8101 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     8161 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     8221 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     8281 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     8341 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     8401 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     8461 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     8521 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     8581 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     8641 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     8701 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc
     8761 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     8821 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     8881 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     8941 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     9001 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     9061 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     9121 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     9181 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     9241 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     9301 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     9361 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     9421 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     9481 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     9541 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     9601 cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta
     9661 aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa
     9721 cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt
     9781 attacaggga cagcagagat ccagtttggt tagtaccggg cccgctctag ccctagattt
     9841 aaatggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     9901 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
     9961 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
    10021 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
    10081 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
    10141 aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg
    10201 gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc
    10261 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
    10321 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
    10381 ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg
    10441 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
    10501 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
    10561 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
    10621 accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg
    10681 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
    10741 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
    10801 cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
    10861 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
    10921 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
    10981 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgaggaatta gcttggtact
    11041 aatacgactc actataggga gacccaagct ggctagccgc ggaatctaga ctgccggatc
    11101 gaattgtcga cgaattcg
//
