LOCUS       sequence-413561-       14138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CLIP donor for integrating CRISPRa (dCas12-miniVPR)-T2A-GFP-P2A to 
            the ACTB locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14138)
  AUTHORS   Chavez M, Rane DA, Chen X, Qi LS
  TITLE     Stable expression of large transgenes via the knock-in of an 
            integrase-deficient lentivirus.
  JOURNAL   Nat Biomed Eng. 2023 May 1. doi: 10.1038/s41551-023-01037-x.
  PUBMED    37127707
REFERENCE   2  (bases 1 to 14138)
  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 Biomed 
            Eng. 2023 May 1. doi: 10.1038/s41551-023-01037-x."
COMMENT     Sequence Label: pSLQ6332 - ACTB CRISPRa CLIP Donor
FEATURES             Location/Qualifiers
     source          1..14138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(830..850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        835..1164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1015..1150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2298..2363
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2493..2513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2938..3072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2975..2994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3029..3048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             3240..3873
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3920..4045
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4542..4775
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4960..5004
                     /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    5289..5406
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             5942..9625
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFRNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIRVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGRGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKRIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIARGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             9632..9658
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             9638..9664
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             9677..9697
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9704..9724
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9761..9910
                     /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             9917..9937
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10070..10426
                     /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             10655..10708
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             10715..11380
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             10715..11380
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             11405..11461
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     LTR             11939..12172
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(12197..12215)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(12198..12215)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(12505..12524)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(12738..12757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12857..12879
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(12917..12935)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        13003..13107
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(13108..13176,13177..13968)
                     /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(13326..13345)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgtg gaatgtgtgt
      841 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
      901 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1081 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1141 tttggaggcc taggcttttg caaaaagctt ggacacaaga caggcttgcg agatatgttt
     1201 gagaatacca ctttatcccg cgtcagggag aggcagtgcg taaaaagacg cggactcatg
     1261 tgaaatactg gtttttagtg cgccagatct ctataatctc gcgcaaccta ttttcccctc
     1321 gaacactttt taagccgtag ataaacaggc tgggacactt cacatgagcg aaaaatacat
     1381 cgtcacctgg gacatgttgc agatccatgc acgtaaactc gcaagccgac tgatgccttc
     1441 tgaacaatgg aaaggcatta ttgccgtaag ccgtggcggt ctgtaccggg tgcgttactg
     1501 gcgcgtgaac tgggtattcg tcatgtcgat accgtttgta tttccagcta cgatcacgac
     1561 aaccagcgcg agcttaaagt gctgaaacgc gcagaaggcg atggcgaagg cttcatcgtt
     1621 attgatgacc tggtggatac cggtggtact gcggttgcga ttcgtgaaat gtatccaaaa
     1681 gcgcactttg tcaccatctt cgcaaaaccg gctggtcgtc cgctggttga tgactatgtt
     1741 gttgatatcc cgcaagatac ctggattgaa cagccgtggg atatgggcgt cgtattcgtc
     1801 ccgccaatct ccggtcgcta atcttttcaa cgcctggcac tgccgggcgt tgttcttttt
     1861 aacttcaggc gggttacaat agtttccagt aagtattctg gaggctgcat ccatgacaca
     1921 ggcaaacctg agcgaaaccc tgttcaaacc ccgctttaaa catcctgaaa cctcgacgct
     1981 agtccgccgc tttaatcacg gcgcacaacc gcctgtgcag tcggcccttg atggtaaaac
     2041 catccctcac tggtatcgca tgattaaccg tctgatgtgg atctggcgcg gcattgaccc
     2101 acgcgaaatc ctcgacgtcc aggcacgtat tgtgatgagc gatgccgaac gtaccgacga
     2161 tgatttatac gatacggtga ttggctaccg tggcggcaac tggatttatg agtgggcccc
     2221 ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag
     2281 agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga
     2341 ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt
     2401 ggaatgcctt taatgaggaa aacctgtttt gctcagaaga aatgccatct agtgatgatg
     2461 aggctactgc tgactctcaa cattctactc ctccaaaaaa gaagagaaag gtagaagacc
     2521 ccaaggactt tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa
     2581 ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa
     2641 ttatggaaaa atattctgta acctttataa gtaggcataa cagttataat cataacatac
     2701 tgttttttct tactccacac aggcatagag tgtctgctat taataactat gctcaaaaat
     2761 tgtgtacctt tagcttttta atttgtaaag gggttaataa ggaatatttg atgtatagtg
     2821 ccttgactag agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2881 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2941 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3001 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3061 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
     3121 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
     3181 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt
     3241 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac
     3301 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
     3361 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
     3421 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     3481 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
     3541 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
     3601 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
     3661 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
     3721 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     3781 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     3841 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     3901 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     3961 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     4021 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     4081 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     4141 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     4201 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     4261 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     4321 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     4381 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     4441 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     4501 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     4561 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     4621 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     4681 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     4741 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     4801 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     4861 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     4921 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     4981 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     5041 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     5101 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     5161 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     5221 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccaaat ggcagtattc
     5281 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     5341 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     5401 aattttcggg tttattacag ggacagcaga gatccagttt ggatcgataa gcttgatatc
     5461 gaattccgcg gcgatatcat catccatggc tgggactcaa ggcgctaact gcgcgtgcgt
     5521 tctggggccc ggggtgccgc ggcctgggct ggggcgaagg cgggctcggc cggaaggggt
     5581 ggggtcgccg cggctcccgg gcgcttgcgc gcacttcctg cccgagccgc tggccgcccg
     5641 agggtgtggc cgctgcgtgc gcgcgcgccg acccggcgct gtttgaaccg ggcggaggcg
     5701 gggctggcgc ccggttggga gggggttggg gcctggcttc ctgccgcgcg ccgcggggac
     5761 gcctccgacc agtgtttgcc ttttatggta ataacgcggc cggcccggct tcctttgtcc
     5821 ccaatctggg cgcgcgccgg cgccccctgg cggcctaagg actcggcgcg ccggaagtgg
     5881 ccagggcggg ggcgacctcg gctcacagcg cgcccggcta ttctcgcaac tgacaggatc
     5941 catgagcaag ctggagaagt ttacaaactg ctactccctg tctaagaccc tgaggttcaa
     6001 ggccatccct gtgggcaaga cccaggagaa catcgacaat aagcggctgc tggtggagga
     6061 cgagaagaga gccgaggatt ataagggcgt gaagaagctg ctggatcgct actatctgtc
     6121 ttttatcaac gacgtgctgc acagcatcaa gctgaagaat ctgaacaatt acatcagcct
     6181 gttccggaag aaaaccagaa ccgagaagga gaataaggag ctggagaacc tggagatcaa
     6241 tctgcggaag gagatcgcca aggccttcaa gggcaacgag ggctacaagt ccctgtttaa
     6301 gaaggatatc atcgagacaa tcctgccaga gttcctggac gataaggacg agatcgccct
     6361 ggtgaacagc ttcaatggct ttaccacagc cttcaccggc ttctttcgta acagagagaa
     6421 tatgttttcc gaggaggcca agagcacatc catcgccttc aggtgtatca acgagaatct
     6481 gacccgctac atctctaata tggacatctt cgagaaggtg gacgccatct ttgataagca
     6541 cgaggtgcag gagatcaagg agaagatcct gaacagcgac tatgatgtgg aggatttctt
     6601 tgagggcgag ttctttaact ttgtgctgac acaggagggc atccgcgtgt ataacgccat
     6661 catcggcggc ttcgtgaccg agagcggcga gaagatcaag ggcctgaacg agtacatcaa
     6721 cctgtataat cagaaaacca agcagaagct gcctaagttt aagccactgt ataagcaggt
     6781 gctgagcgat cgggagtctc tgagcttcta cggcaggggc tatacatccg atgaggaggt
     6841 gctggaggtg tttagaaaca ccctgaacaa gaacagcgag atcttcagct ccatcaagaa
     6901 gctggagaag ctgttcaaga attttgacga gtactctagc gccggcatct ttgtgaagaa
     6961 cggccccgcc atcagcacaa tctccaagcg tatcttcggc gagtggaacg tgatccggga
     7021 caagtggaat gccgagtatg acgatatcca cctgaagaag aaggccgtgg tgaccgagaa
     7081 gtacgaggac gatcggagaa agtccttcaa gaagatcggc tccttttctc tggagcagct
     7141 gcaggagtac gccgacgccg atctgtctgt ggtggagaag ctgaaggaga tcatcatcca
     7201 gaaggtggat gagatctaca aggtgtatgg ctcctctgag aagctgttcg acgccgattt
     7261 tgtgctggag aagagcctga agaagaacga cgccgtggtg gccatcatga aggacctgct
     7321 ggattctgtg aagagcttcg agaattacat caaggccttc tttggcgagg gcaaggagac
     7381 aaacagggac gagtccttct atggcgattt tgtgctggcc tacgacatcc tgctgaaggt
     7441 ggaccacatc tacgatgcca tccgcaatta tgtgacccag aagccctact ctaaggataa
     7501 gttcaagctg tattttcaga accctcagtt catgggcggc tgggacaagg ataaggagac
     7561 agactatcgg gccaccatcc tgagatacgg ctccaagtac tatctggcca tcatggataa
     7621 gaagtacgcc aagtgcctgc agaagatcga caaggacgat gtgaacggca attacgagaa
     7681 gatcaactat aagctgctgc ccggccctaa taagatgctg ccaaaggtgt tcttttctaa
     7741 gaagtggatg gcctactata accccagcga ggacatccag aagatctaca agaatggcac
     7801 attcaagaag ggcgatatgt ttaacctgaa tgactgtcac aagctgatcg acttctttaa
     7861 ggatagcatc tcccggtatc caaagtggtc caatgcctac gatttcaact tttctgagac
     7921 agagaagtat aaggacatcg ccggctttta cagagaggtg gaggagcagg gctataaggt
     7981 gagcttcgag tctgccagca agaaggaggt ggataagctg gtggaggagg gcaagctgta
     8041 tatgttccag atctataaca aggacttttc cgataagtct cacggcacac ccaatctgca
     8101 caccatgtac ttcaagctgc tgtttgacga gaacaatcac ggacagatca ggctgagcgg
     8161 aggagcagag ctgttcatga ggcgcgcctc cctgaagaag gaggagctgg tggtgcaccc
     8221 agccaactcc cctatcgcca acaagaatcc agataatccc aagaaaacca caaccctgtc
     8281 ctacgacgtg tataaggata agaggttttc tgaggaccag tacgagctgc acatcccaat
     8341 cgccatcaat aagtgcccca agaacatctt caagatcaat acagaggtgc gcgtgctgct
     8401 gaagcacgac gataacccct atgtgatcgg catcgccagg ggcgagcgca atctgctgta
     8461 tatcgtggtg gtggacggca agggcaacat cgtggagcag tattccctga acgagatcat
     8521 caacaacttc aacggcatca ggatcaagac agattaccac tctctgctgg acaagaagga
     8581 gaaggagagg ttcgaggccc gccagaactg gacctccatc gagaatatca aggagctgaa
     8641 ggccggctat atctctcagg tggtgcacaa gatctgcgag ctggtggaga agtacgatgc
     8701 cgtgatcgcc ctggaggacc tgaactctgg ctttaagaat agccgcgtga aggtggagaa
     8761 gcaggtgtat cagaagttcg agaagatgct gatcgataag ctgaactaca tggtggacaa
     8821 gaagtctaat ccttgtgcaa caggcggcgc cctgaagggc tatcagatca ccaataagtt
     8881 cgagagcttt aagtccatgt ctacccagaa cggcttcatc ttttacatcc ctgcctggct
     8941 gacatccaag atcgatccat ctaccggctt tgtgaacctg ctgaaaacca agtataccag
     9001 catcgccgat tccaagaagt tcatcagctc ctttgacagg atcatgtacg tgcccgagga
     9061 ggatctgttc gagtttgccc tggactataa gaacttctct cgcacagacg ccgattacat
     9121 caagaagtgg aagctgtact cctacggcaa ccggatcaga atcttccgga atcctaagaa
     9181 gaacaacgtg ttcgactggg aggaggtgtg cctgaccagc gcctataagg agctgttcaa
     9241 caagtacggc atcaattatc agcagggcga tatcagagcc ctgctgtgcg agcagtccga
     9301 caaggccttc tactctagct ttatggccct gatgagcctg atgctgcaga tgcggaacag
     9361 catcacaggc cgcaccgacg tggattttct gatcagccct gtgaagaact ccgacggcat
     9421 cttctacgat agccggaact atgaggccca ggagaatgcc atcctgccaa agaacgccga
     9481 cgccaatggc gcctataaca tcgccagaaa ggtgctgtgg gccatcggcc agttcaagaa
     9541 ggccgaggac gagaagctgg ataaggtgaa gatcgccatc tctaacaagg agtggctgga
     9601 gtacgcccag accagcgtga agcacggatc ctatccctat gacgtgcccg attatgccag
     9661 cctgggcagc ggctccccca agaaaaaacg caaggtggaa gatcctaaga aaaagcggaa
     9721 agtggacggc attggtagtg ggagcaacgg cagcagcctc gatgctttag acgattttga
     9781 cttagatatg cttggttcag acgcgttaga cgacttcgac ctagacatgt taggctcaga
     9841 tgcattggac gacttcgatt tagatatgtt gggctccgat gccctagatg actttgatct
     9901 agatatgcta ggtagtccca aaaagaagag gaaagtggga tccgtccttc cgcaagcgcc
     9961 tgccccagcg cctgcaccgg caatggtgtc cgccctcgca caggcccctg cgcccgtccc
    10021 cgtgctcgcg cctggaccgc cccaggcggt cgctccaccg gctccgaagc cgacgcaggc
    10081 cggagaggga acactctccg aagcacttct tcaactccag tttgatgacg aggatcttgg
    10141 agcactcctt ggaaactcga cagaccctgc ggtgtttacc gacctcgcgt cagtagataa
    10201 ctccgaattt cagcagcttt tgaaccaggg tatcccggtc gcgccacata caacggagcc
    10261 catgttgatg gaataccccg aagcaatcac gagacttgtg acgggagcgc agcggcctcc
    10321 cgatcccgca cccgcacctt tgggggcacc tggcctccct aacggacttt tgagcggcga
    10381 cgaggatttc tcctccatcg ccgatatgga tttctcagcc ttgctgtcag gctctggcag
    10441 cggcagcgac ctttcccatc cgcccccaag gggccatctg gatgagctga caaccacact
    10501 tgagtccatg accgaggatc tgaacctgga ctcacccctg accccggaat tgaacgagat
    10561 tctggatacc ttcctgaacg acgagtgcct cttgcatgcc atgcatatca gcacaggact
    10621 gtccatcttc gacacatctc tgtttggatc gggagagggc agaggaagtc tgctaacatg
    10681 cggtgacgtc gaggagaatc ctggcccagg atccatgccc gccatgaaga tcgagtgccg
    10741 catcaccggc accctgaacg gcgtggagtt cgagctggtg ggcggcggag agggcacccc
    10801 cgagcagggc cgcatgacca acaagatgaa gagcaccaaa ggcgccctga ccttcagccc
    10861 ctacctgctg agccacgtga tgggctacgg cttctaccac ttcggcacct accccagcgg
    10921 ctacgagaac cccttcctgc acgccatcaa caacggcggc tacaccaaca cccgcatcga
    10981 gaagtacgag gacggcggcg tgctgcacgt gagcttcagc taccgctacg aggccggccg
    11041 cgtgatcggc gacttcaagg tggtgggcac cggcttcccc gaggacagcg tgatcttcac
    11101 cgacaagatc atccgcagca acgccaccgt ggagcacctg caccccatgg gcgataacgt
    11161 gctggtgggc agcttcgccc gcaccttcag cctgcgcgac ggcggctact acagcttcgt
    11221 ggtggacagc cacatgcact tcaagagcgc catccacccc agcatcctgc agaacggggg
    11281 ccccatgttc gccttccgcc gcgtggagga gctgcacagc aacaccgagc tgggcatcgt
    11341 ggagtaccag cacgccttca agacccccat cgccttcgcc agatctcgag tccccgggag
    11401 cggagctact aacttcagcc tgctgaagca ggctggagac gtggaggaga accctggacc
    11461 tgccggctcc ggtaccgatg atgatatcgc agcgctcgtc gtcgacaacg gctccggcat
    11521 gtgcaaggcc ggcttcgcgg gcgacgatgc cccccgggcc gtcttcccct ccatcgtggg
    11581 gcgccccagg gaccaggtag gggagctggc tgggtggggc agccccggga gcgggcggga
    11641 ggcaagggcg ctttctctgc acaggagcct cccggtttcc ggggtggggg ctgcgcccgt
    11701 gctcagggct tcttgtcctt tccttcccag ggcgtgatgg tgggcatggg tcagaaggat
    11761 tcctatgtgg gcgacgaggc ccagagcaag agaggcatcc tcaccctgaa gtaccccatc
    11821 gccatggatg atgatatcgc cggcctcgag ggaattaatt cgagctcggt acctttaaga
    11881 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
    11941 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
    12001 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
    12061 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
    12121 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcatctag
    12181 aattaattcc gtgtattcta tagtgtcacc taaatcgtat gtgtatgata cataaggtta
    12241 tgtattaatt gtagccgcgt tctaacgaca atatgtacaa gcctaattgt gtagcatctg
    12301 gcttactgaa gcagacccta tcatctctct cgtaaactgc cgtcagagtc ggtttggttg
    12361 gacgaacctt ctgagtttct ggtaacgccg tcccgcaccc ggaaatggtc agcgaaccaa
    12421 tcagcagggt catcgctagc cagatcctct acgccggacg catcgtggcc ggcatcaccg
    12481 gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg
    12541 ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg
    12601 ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca
    12661 acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc
    12721 gaatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
    12781 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
    12841 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
    12901 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
    12961 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
    13021 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
    13081 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
    13141 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
    13201 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
    13261 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
    13321 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
    13381 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
    13441 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
    13501 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
    13561 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
    13621 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
    13681 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
    13741 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
    13801 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
    13861 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
    13921 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
    13981 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
    14041 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
    14101 tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
