LOCUS       sequence-148551-       10438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recruits VPR to compatible gRNAs for transcriptional activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10438)
  AUTHORS   Chavez A, Tuttle M, Pruitt BW, Ewen-Campen B, Chari R, Ter-Ovanesyan
            D, Haque SJ, Cecchi RJ, Kowal EJ, Buchthal J, Housden BE, Perrimon 
            N, Collins JJ, Church G
  TITLE     Comparison of Cas9 activators in multiple species.
  JOURNAL   Nat Methods. 2016 May 23. doi: 10.1038/nmeth.3871.
  PUBMED    27214048
REFERENCE   2  (bases 1 to 10438)
  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 
            Methods. 2016 May 23. doi: 10.1038/nmeth.3871."
COMMENT     Sequence Label: pLEX_307_ms2_VPR
FEATURES             Location/Qualifiers
     source          1..10438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..1274
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          327..1265
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1222..1242
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1294..1318
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1330..1350
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1360..1383
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1774..1923
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             1948..1968
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2398..2754
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             2773..3342
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     protein_bind    complement(3347..3371)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3379..3420
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3471..4059
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3524..3544)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3942..3953)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4131..4364
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4442..4563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4479..4498)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4533..4552
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4603..4738
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4758..4777)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4759..4777)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4787..4804)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4787..4803)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4796..4818)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4945..5400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5032..5051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5242..5263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5426..5530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5531..5599,5600..6391)
                     /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(5749..5768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6562..7150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7051..7070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7304..7321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7438..7459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7474..7479,7480..7497,7498..7504)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7512..7528
                     /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     7517..7539
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7536..7552
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7536..7552
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7571..7591
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7573..7591
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7619..7845
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7846..8026
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8073..8198
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8691..8924
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9109..9153
                     /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"
     promoter        9384..9594
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        9406..9602
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9453..9588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             9634..10236
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(9634..9653)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10130..10150
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10279..10396
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctcgaggcct gcaggtgcaa agatggataa agttttaaac agagaggaat ctttgcagct
       61 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
      121 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
      181 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      241 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      301 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
      361 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
      421 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
      481 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
      541 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
      601 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
      661 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
      721 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
      781 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
      841 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
      901 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
      961 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     1021 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     1081 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     1141 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     1201 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     1261 ttcaggtgtc gtgaggctag catcgattga tcaacaagtt tgtacaaaaa agcaggctac
     1321 aaaatgggac ctaagaaaaa gaggaaggtg gcggccgctg actacaagga tgacgacgat
     1381 aaatctagaa tggcttctaa ctttactcag ttcgttctcg tcgacaatgg cggaactggc
     1441 gacgtgactg tcgccccaag caacttcgct aacgggatcg ctgaatggat cagctctaac
     1501 tcgcgttcac aggcttacaa agtaacctgt agcgttcgtc agagctctgc gcagaatcgc
     1561 aaatacacca tcaaagtcga ggtgcctaaa ggcgcctggc gttcgtactt aaatatggaa
     1621 ctaaccattc caattttcgc cacgaattcc gactgcgagc ttattgttaa ggcaatgcaa
     1681 ggtctcctaa aagatggaaa cccgattccc tcagcaatcg cagcaaactc cggcatctac
     1741 gaggccagcg aggccagcgg ttccggacgg gctgacgcat tggacgattt tgatctggat
     1801 atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc ggatgccctt
     1861 gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga cctggacatg
     1921 ctgattaact ctagaagttc cggatctccg aaaaagaaac gcaaagttgg tagccagtac
     1981 ctgcccgaca ccgacgaccg gcaccggatc gaggaaaagc ggaagcggac ctacgagaca
     2041 ttcaagagca tcatgaagaa gtcccccttc agcggcccca ccgaccctag acctccacct
     2101 agaagaatcg ccgtgcccag cagatccagc gccagcgtgc caaaacctgc cccccagcct
     2161 taccccttca ccagcagcct gagcaccatc aactacgacg agttccctac catggtgttc
     2221 cccagcggcc agatctctca ggcctctgct ctggctccag cccctcctca ggtgctgcct
     2281 caggctcctg ctcctgcacc agctccagcc atggtgtctg cactggctca ggcaccagca
     2341 cccgtgcctg tgctggctcc tggacctcca caggctgtgg ctccaccagc ccctaaacct
     2401 acacaggccg gcgagggcac actgtctgaa gctctgctgc agctgcagtt cgacgacgag
     2461 gatctgggag ccctgctggg aaacagcacc gatcctgccg tgttcaccga cctggccagc
     2521 gtggacaaca gcgagttcca gcagctgctg aaccagggca tccctgtggc ccctcacacc
     2581 accgagccca tgctgatgga ataccccgag gccatcaccc ggctcgtgac aggcgctcag
     2641 aggcctcctg atccagctcc tgcccctctg ggagcaccag gcctgcctaa tggactgctg
     2701 tctggcgacg aggacttcag ctctatcgcc gatatggatt tctcagcctt gctgggctct
     2761 ggcagcggca gccgggattc cagggaaggg atgtttttgc cgaagcctga ggccggctcc
     2821 gctattagtg acgtgtttga gggccgcgag gtgtgccagc caaaacgaat ccggccattt
     2881 catcctccag gaagtccatg ggccaaccgc ccactccccg ccagcctcgc accaacacca
     2941 accggtccag tacatgagcc agtcgggtca ctgaccccgg caccagtccc tcagccactg
     3001 gatccagcgc ccgcagtgac tcccgaggcc agtcacctgt tggaggatcc cgatgaagag
     3061 acgagccagg ctgtcaaagc ccttcgggag atggccgata ctgtgattcc ccagaaggaa
     3121 gaggctgcaa tctgtggcca aatggacctt tcccatccgc ccccaagggg ccatctggat
     3181 gagctgacaa ccacacttga gtccatgacc gaggatctga acctggactc acccctgacc
     3241 ccggaattga acgagattct ggataccttc ctgaacgacg agtgcctctt gcatgccatg
     3301 catatcagca caggactgtc catcttcgac acatctctgt tttgaaaccc agctttcttg
     3361 tacaaagtgg ttgatatcgg taagcctatc cctaaccctc tcctcggtct cgattctacg
     3421 tagtaatgaa ctagtaccgg ttaagtcgac aatcaacgcg ttaagtcgac aatcaacctc
     3481 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3541 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3601 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3661 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3721 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3781 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3841 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     3901 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3961 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     4021 ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac
     4081 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     4141 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     4201 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     4261 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     4321 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtacgt atagtagttc
     4381 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     4441 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4501 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4561 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg
     4621 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     4681 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4741 gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     4801 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     4861 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     4921 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4981 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5041 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5101 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5161 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5221 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5281 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5341 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     5401 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5461 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5521 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5581 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5641 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5701 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5761 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5821 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5881 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5941 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6001 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6061 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6121 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6181 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6241 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6301 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6361 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6421 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6481 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6541 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6601 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6661 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6721 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6781 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6841 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6901 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6961 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7021 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7081 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7141 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7201 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7261 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7321 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7381 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     7441 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     7501 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     7561 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg
     7621 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     7681 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     7741 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     7801 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     7861 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     7921 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     7981 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     8041 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     8101 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     8161 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     8221 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     8281 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     8341 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     8401 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     8461 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     8521 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     8581 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     8641 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     8701 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     8761 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     8821 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     8881 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     8941 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     9001 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     9061 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     9121 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     9181 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     9241 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     9301 acccacctcc caaccccgag gggacccatg cattgcatct caattagtca gcaaccaggt
     9361 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcgt ctcaattagt
     9421 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     9481 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     9541 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     9601 aaaagctttc tagaggtacc accggagctt accatgaccg agtacaagcc cacggtgcgc
     9661 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     9721 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     9781 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     9841 ggcgccgccg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     9901 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     9961 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
    10021 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
    10081 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
    10141 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
    10201 acctggtgca tgacccgcaa gcccggtgcc agttaacgaa ttctagatct tgagacaaat
    10261 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
    10321 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
    10381 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccacttt ggcgccgg
//
