LOCUS       sequence-363081-       11363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG-hyperdCas12a-HA-miniVPR-mCherry-T2A-HygroR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11363)
  AUTHORS   Guo LY, Bian J, Davis AE, Liu P, Kempton HR, Zhang X, Chemparathy A,
            Gu B, Lin X, Rane DA, Xu X, Jamiolkowski RM, Hu Y, Wang S, Qi LS
  TITLE     Multiplexed genome regulation in vivo with hyper-efficient Cas12a.
  JOURNAL   Nat Cell Biol. 2022 Apr;24(4):590-600. doi: 
            10.1038/s41556-022-00870-7. Epub 2022 Apr 12.
  PUBMED    35414015
REFERENCE   2  (bases 1 to 11363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41556-022-00870-7"; journalName: "Nat Cell Biol"; date: 
            "2022-04"; volume: "24"; issue: "4"; pages: "590-600"
COMMENT     Sequence Label: pSLQ10844
FEATURES             Location/Qualifiers
     source          1..11363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1776..5459
                     /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             5466..5492
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5472..5498
                     /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             5511..5531
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5538..5558
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5595..5744
                     /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             5751..5771
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5904..6260
                     /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             6495..7202
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6640..6658)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6908..6927
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7212..7265
                     /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             7266..8291
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             7266..8291
                     /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"
     primer_bind     complement(8385..8404)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8450..8505
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8504..8523)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8866..8882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8866..8882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8879..8901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8890..8906
                     /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)."
     promoter        complement(join(8914..8920,8921..8938,8939..8944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8959..8980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9039..9234
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9085..9220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    9240..9374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9277..9296)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9331..9350
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9442..9459)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9613..10201)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9693..9712)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10372..11163,11164..11232))
                     /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     10995..11014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11233..11337)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggnsgsg cgccrggsgg ggsggggsgg ggsgrggggs
      541 ggggsggggs grggsggrgr ggkgsggsgg crgccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggkgggggtg ccgggcgggg
     1261 cggggccgcc ycgggccggg grgggctcgg gggrggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttctgcagtc
     1741 gacggtaccg cgggcccggg atccaccggg ccaccatgag caagctggag aagtttacaa
     1801 actgctactc cctgtctaag accctgaggt tcaaggccat ccctgtgggc aagacccagg
     1861 agaacatcga caataagcgg ctgctggtgg aggacgagaa gagagccgag gattataagg
     1921 gcgtgaagaa gctgctggat cgctactatc tgtcttttat caacgacgtg ctgcacagca
     1981 tcaagctgaa gaatctgaac aattacatca gcctgttccg gaagaaaacc agaaccgaga
     2041 aggagaataa ggagctggag aacctggaga tcaatctgcg gaaggagatc gccaaggcct
     2101 tcaagggcaa cgagggctac aagtccctgt ttaagaagga tatcatcgag acaatcctgc
     2161 cagagttcct ggacgataag gacgagatcg ccctggtgaa cagcttcaat ggctttacca
     2221 cagccttcac cggcttcttt cgtaacagag agaatatgtt ttccgaggag gccaagagca
     2281 catccatcgc cttcaggtgt atcaacgaga atctgacccg ctacatctct aatatggaca
     2341 tcttcgagaa ggtggacgcc atctttgata agcacgaggt gcaggagatc aaggagaaga
     2401 tcctgaacag cgactatgat gtggaggatt tctttgaggg cgagttcttt aactttgtgc
     2461 tgacacagga gggcatccgc gtgtataacg ccatcatcgg cggcttcgtg accgagagcg
     2521 gcgagaagat caagggcctg aacgagtaca tcaacctgta taatcagaaa accaagcaga
     2581 agctgcctaa gtttaagcca ctgtataagc aggtgctgag cgatcgggag tctctgagct
     2641 tctacggcag gggctataca tccgatgagg aggtgctgga ggtgtttaga aacaccctga
     2701 acaagaacag cgagatcttc agctccatca agaagctgga gaagctgttc aagaattttg
     2761 acgagtactc tagcgccggc atctttgtga agaacggccc cgccatcagc acaatctcca
     2821 agcgtatctt cggcgagtgg aacgtgatcc gggacaagtg gaatgccgag tatgacgata
     2881 tccacctgaa gaagaaggcc gtggtgaccg agaagtacga ggacgatcgg agaaagtcct
     2941 tcaagaagat cggctccttt tctctggagc agctgcagga gtacgccgac gccgatctgt
     3001 ctgtggtgga gaagctgaag gagatcatca tccagaaggt ggatgagatc tacaaggtgt
     3061 atggctcctc tgagaagctg ttcgacgccg attttgtgct ggagaagagc ctgaagaaga
     3121 acgacgccgt ggtggccatc atgaaggacc tgctggattc tgtgaagagc ttcgagaatt
     3181 acatcaaggc cttctttggc gagggcaagg agacaaacag ggacgagtcc ttctatggcg
     3241 attttgtgct ggcctacgac atcctgctga aggtggacca catctacgat gccatccgca
     3301 attatgtgac ccagaagccc tactctaagg ataagttcaa gctgtatttt cagaaccctc
     3361 agttcatggg cggctgggac aaggataagg agacagacta tcgggccacc atcctgagat
     3421 acggctccaa gtactatctg gccatcatgg ataagaagta cgccaagtgc ctgcagaaga
     3481 tcgacaagga cgatgtgaac ggcaattacg agaagatcaa ctataagctg ctgcccggcc
     3541 ctaataagat gctgccaaag gtgttctttt ctaagaagtg gatggcctac tataacccca
     3601 gcgaggacat ccagaagatc tacaagaatg gcacattcaa gaagggcgat atgtttaacc
     3661 tgaatgactg tcacaagctg atcgacttct ttaaggatag catctcccgg tatccaaagt
     3721 ggtccaatgc ctacgatttc aacttttctg agacagagaa gtataaggac atcgccggct
     3781 tttacagaga ggtggaggag cagggctata aggtgagctt cgagtctgcc agcaagaagg
     3841 aggtggataa gctggtggag gagggcaagc tgtatatgtt ccagatctat aacaaggact
     3901 tttccgataa gtctcacggc acacccaatc tgcacaccat gtacttcaag ctgctgtttg
     3961 acgagaacaa tcacggacag atcaggctga gcggaggagc agagctgttc atgaggcgcg
     4021 cctccctgaa gaaggaggag ctggtggtgc acccagccaa ctcccctatc gccaacaaga
     4081 atccagataa tcccaagaaa accacaaccc tgtcctacga cgtgtataag gataagaggt
     4141 tttctgagga ccagtacgag ctgcacatcc caatcgccat caataagtgc cccaagaaca
     4201 tcttcaagat caatacagag gtgcgcgtgc tgctgaagca cgacgataac ccctatgtga
     4261 tcggcatcgc caggggcgag cgcaatctgc tgtatatcgt ggtggtggac ggcaagggca
     4321 acatcgtgga gcagtattcc ctgaacgaga tcatcaacaa cttcaacggc atcaggatca
     4381 agacagatta ccactctctg ctggacaaga aggagaagga gaggttcgag gcccgccaga
     4441 actggacctc catcgagaat atcaaggagc tgaaggccgg ctatatctct caggtggtgc
     4501 acaagatctg cgagctggtg gagaagtacg atgccgtgat cgccctggag gacctgaact
     4561 ctggctttaa gaatagccgc gtgaaggtgg agaagcaggt gtatcagaag ttcgagaaga
     4621 tgctgatcga taagctgaac tacatggtgg acaagaagtc taatccttgt gcaacaggcg
     4681 gcgccctgaa gggctatcag atcaccaata agttcgagag ctttaagtcc atgtctaccc
     4741 agaacggctt catcttttac atccctgcct ggctgacatc caagatcgat ccatctaccg
     4801 gctttgtgaa cctgctgaaa accaagtata ccagcatcgc cgattccaag aagttcatca
     4861 gctcctttga caggatcatg tacgtgcccg aggaggatct gttcgagttt gccctggact
     4921 ataagaactt ctctcgcaca gacgccgatt acatcaagaa gtggaagctg tactcctacg
     4981 gcaaccggat cagaatcttc cggaatccta agaagaacaa cgtgttcgac tgggaggagg
     5041 tgtgcctgac cagcgcctat aaggagctgt tcaacaagta cggcatcaat tatcagcagg
     5101 gcgatatcag agccctgctg tgcgagcagt ccgacaaggc cttctactct agctttatgg
     5161 ccctgatgag cctgatgctg cagatgcgga acagcatcac aggccgcacc gacgtggatt
     5221 ttctgatcag ccctgtgaag aactccgacg gcatcttcta cgatagccgg aactatgagg
     5281 cccaggagaa tgccatcctg ccaaagaacg ccgacgccaa tggcgcctat aacatcgcca
     5341 gaaaggtgct gtgggccatc ggccagttca agaaggccga ggacgagaag ctggataagg
     5401 tgaagatcgc catctctaac aaggagtggc tggagtacgc ccagaccagc gtgaagcacg
     5461 gatcctatcc ctatgacgtg cccgattatg ccagcctggg cagcggctcc cccaagaaaa
     5521 aacgcaaggt ggaagatcct aagaaaaagc ggaaagtgga cggcattggt agtgggagca
     5581 acggcagcag cctcgatgct ttagacgatt ttgacttaga tatgcttggt tcagacgcgt
     5641 tagacgactt cgacctagac atgttaggct cagatgcatt ggacgacttc gatttagata
     5701 tgttgggctc cgatgcccta gatgactttg atctagatat gctaggtagt cccaaaaaga
     5761 agaggaaagt gggatccgtc cttccgcaag cgcctgcccc agcgcctgca ccggcaatgg
     5821 tgtccgccct cgcacaggcc cctgcgcccg tccccgtgct cgcgcctgga ccgccccagg
     5881 cggtcgctcc accggctccg aagccgacgc aggccggaga gggaacactc tccgaagcac
     5941 ttcttcaact ccagtttgat gacgaggatc ttggagcact ccttggaaac tcgacagacc
     6001 ctgcggtgtt taccgacctc gcgtcagtag ataactccga atttcagcag cttttgaacc
     6061 agggtatccc ggtcgcgcca catacaacgg agcccatgtt gatggaatac cccgaagcaa
     6121 tcacgagact tgtgacggga gcgcagcggc ctcccgatcc cgcacccgca cctttggggg
     6181 cacctggcct ccctaacgga cttttgagcg gcgacgagga tttctcctcc atcgccgata
     6241 tggatttctc agccttgctg tcaggctctg gcagcggcag cgacctttcc catccgcccc
     6301 caaggggcca tctggatgag ctgacaacca cacttgagtc catgaccgag gatctgaacc
     6361 tggactcacc cctgaccccg gaattgaacg agattctgga taccttcctg aacgacgagt
     6421 gcctcttgca tgccatgcat atcagcacag gactgtccat cttcgacaca tctctgtttg
     6481 gcagcagcgg atccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     6541 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     6601 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     6661 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     6721 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     6781 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     6841 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     6901 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     6961 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     7021 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     7081 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     7141 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     7201 agggatccgg cgagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg
     7261 gccctatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     7321 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     7381 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     7441 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     7501 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     7561 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     7621 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     7681 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     7741 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     7801 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     7861 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     7921 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     7981 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     8041 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     8101 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     8161 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     8221 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     8281 caaaggaata ggtacaagta aggccgcact cctcaggtgc aggctgccta tcagaaggtg
     8341 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc
     8401 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     8461 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     8521 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     8581 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa
     8641 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     8701 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     8761 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     8821 ttatggagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     8881 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     8941 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     9001 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     9061 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     9121 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     9181 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     9241 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     9301 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     9361 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     9421 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     9481 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     9541 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     9601 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     9661 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     9721 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     9781 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     9841 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     9901 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     9961 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
    10021 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
    10081 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
    10141 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
    10201 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
    10261 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    10321 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    10381 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    10441 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    10501 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
    10561 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    10621 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    10681 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    10741 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    10801 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    10861 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    10921 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    10981 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    11041 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    11101 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    11161 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    11221 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    11281 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    11341 atttccccga aaagtgccac ctg
//
