LOCUS       sequence-363077-       11362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG-dCas12a(WT)-HA-miniVPR-mCherry-T2A-HygroR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11362)
  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 11362)
  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: pSLQ10333
FEATURES             Location/Qualifiers
     source          1..11362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1775..5458
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIARGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             5465..5491
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5471..5497
                     /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             5510..5530
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5537..5557
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5594..5743
                     /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             5750..5770
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5903..6259
                     /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             6494..7201
                     /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(6639..6657)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6907..6926
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7211..7264
                     /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             7265..8290
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             7265..8290
                     /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(8384..8403)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8449..8504
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8503..8522)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8865..8881)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8865..8881)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8878..8900)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8889..8905
                     /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(8913..8919,8920..8937,8938..8943))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8958..8979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9038..9233
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9084..9219
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    9239..9373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9276..9295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9330..9349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9441..9458)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9612..10200)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9692..9711)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10371..11162,11163..11231))
                     /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     10994..11013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11232..11336)
                     /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 gggggggsgc gccrggsggg gsggggsggg gsgrggggsg
      541 gggsggggsg rggsggrgrg gkgsggsggc rgccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gkggcggcag gkgggggtgc cgggcggggc
     1261 ggggccgccy cgggccgggg rgggckcggg ggrggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tctgcagtcg
     1741 acggtaccgc gggcccggga tccaccgggc caccatgagc aagctggaga agtttacaaa
     1801 ctgctactcc ctgtctaaga ccctgaggtt caaggccatc cctgtgggca agacccagga
     1861 gaacatcgac aataagcggc tgctggtgga ggacgagaag agagccgagg attataaggg
     1921 cgtgaagaag ctgctggatc gctactatct gtcttttatc aacgacgtgc tgcacagcat
     1981 caagctgaag aatctgaaca attacatcag cctgttccgg aagaaaacca gaaccgagaa
     2041 ggagaataag gagctggaga acctggagat caatctgcgg aaggagatcg ccaaggcctt
     2101 caagggcaac gagggctaca agtccctgtt taagaaggat atcatcgaga caatcctgcc
     2161 agagttcctg gacgataagg acgagatcgc cctggtgaac agcttcaatg gctttaccac
     2221 agccttcacc ggcttctttg ataacagaga gaatatgttt tccgaggagg ccaagagcac
     2281 atccatcgcc ttcaggtgta tcaacgagaa tctgacccgc tacatctcta atatggacat
     2341 cttcgagaag gtggacgcca tctttgataa gcacgaggtg caggagatca aggagaagat
     2401 cctgaacagc gactatgatg tggaggattt ctttgagggc gagttcttta actttgtgct
     2461 gacacaggag ggcatcgacg tgtataacgc catcatcggc ggcttcgtga ccgagagcgg
     2521 cgagaagatc aagggcctga acgagtacat caacctgtat aatcagaaaa ccaagcagaa
     2581 gctgcctaag tttaagccac tgtataagca ggtgctgagc gatcgggagt ctctgagctt
     2641 ctacggcgag ggctatacat ccgatgagga ggtgctggag gtgtttagaa acaccctgaa
     2701 caagaacagc gagatcttca gctccatcaa gaagctggag aagctgttca agaattttga
     2761 cgagtactct agcgccggca tctttgtgaa gaacggcccc gccatcagca caatctccaa
     2821 ggatatcttc ggcgagtgga acgtgatccg ggacaagtgg aatgccgagt atgacgatat
     2881 ccacctgaag aagaaggccg tggtgaccga gaagtacgag gacgatcgga gaaagtcctt
     2941 caagaagatc ggctcctttt ctctggagca gctgcaggag tacgccgacg ccgatctgtc
     3001 tgtggtggag aagctgaagg agatcatcat ccagaaggtg gatgagatct acaaggtgta
     3061 tggctcctct gagaagctgt tcgacgccga ttttgtgctg gagaagagcc tgaagaagaa
     3121 cgacgccgtg gtggccatca tgaaggacct gctggattct gtgaagagct tcgagaatta
     3181 catcaaggcc ttctttggcg agggcaagga gacaaacagg gacgagtcct tctatggcga
     3241 ttttgtgctg gcctacgaca tcctgctgaa ggtggaccac atctacgatg ccatccgcaa
     3301 ttatgtgacc cagaagccct actctaagga taagttcaag ctgtattttc agaaccctca
     3361 gttcatgggc ggctgggaca aggataagga gacagactat cgggccacca tcctgagata
     3421 cggctccaag tactatctgg ccatcatgga taagaagtac gccaagtgcc tgcagaagat
     3481 cgacaaggac gatgtgaacg gcaattacga gaagatcaac tataagctgc tgcccggccc
     3541 taataagatg ctgccaaagg tgttcttttc taagaagtgg atggcctact ataaccccag
     3601 cgaggacatc cagaagatct acaagaatgg cacattcaag aagggcgata tgtttaacct
     3661 gaatgactgt cacaagctga tcgacttctt taaggatagc atctcccggt atccaaagtg
     3721 gtccaatgcc tacgatttca acttttctga gacagagaag tataaggaca tcgccggctt
     3781 ttacagagag gtggaggagc agggctataa ggtgagcttc gagtctgcca gcaagaagga
     3841 ggtggataag ctggtggagg agggcaagct gtatatgttc cagatctata acaaggactt
     3901 ttccgataag tctcacggca cacccaatct gcacaccatg tacttcaagc tgctgtttga
     3961 cgagaacaat cacggacaga tcaggctgag cggaggagca gagctgttca tgaggcgcgc
     4021 ctccctgaag aaggaggagc tggtggtgca cccagccaac tcccctatcg ccaacaagaa
     4081 tccagataat cccaagaaaa ccacaaccct gtcctacgac gtgtataagg ataagaggtt
     4141 ttctgaggac cagtacgagc tgcacatccc aatcgccatc aataagtgcc ccaagaacat
     4201 cttcaagatc aatacagagg tgcgcgtgct gctgaagcac gacgataacc cctatgtgat
     4261 cggcatcgcc aggggcgagc gcaatctgct gtatatcgtg gtggtggacg gcaagggcaa
     4321 catcgtggag cagtattccc tgaacgagat catcaacaac ttcaacggca tcaggatcaa
     4381 gacagattac cactctctgc tggacaagaa ggagaaggag aggttcgagg cccgccagaa
     4441 ctggacctcc atcgagaata tcaaggagct gaaggccggc tatatctctc aggtggtgca
     4501 caagatctgc gagctggtgg agaagtacga tgccgtgatc gccctggagg acctgaactc
     4561 tggctttaag aatagccgcg tgaaggtgga gaagcaggtg tatcagaagt tcgagaagat
     4621 gctgatcgat aagctgaact acatggtgga caagaagtct aatccttgtg caacaggcgg
     4681 cgccctgaag ggctatcaga tcaccaataa gttcgagagc tttaagtcca tgtctaccca
     4741 gaacggcttc atcttttaca tccctgcctg gctgacatcc aagatcgatc catctaccgg
     4801 ctttgtgaac ctgctgaaaa ccaagtatac cagcatcgcc gattccaaga agttcatcag
     4861 ctcctttgac aggatcatgt acgtgcccga ggaggatctg ttcgagtttg ccctggacta
     4921 taagaacttc tctcgcacag acgccgatta catcaagaag tggaagctgt actcctacgg
     4981 caaccggatc agaatcttcc ggaatcctaa gaagaacaac gtgttcgact gggaggaggt
     5041 gtgcctgacc agcgcctata aggagctgtt caacaagtac ggcatcaatt atcagcaggg
     5101 cgatatcaga gccctgctgt gcgagcagtc cgacaaggcc ttctactcta gctttatggc
     5161 cctgatgagc ctgatgctgc agatgcggaa cagcatcaca ggccgcaccg acgtggattt
     5221 tctgatcagc cctgtgaaga actccgacgg catcttctac gatagccgga actatgaggc
     5281 ccaggagaat gccatcctgc caaagaacgc cgacgccaat ggcgcctata acatcgccag
     5341 aaaggtgctg tgggccatcg gccagttcaa gaaggccgag gacgagaagc tggataaggt
     5401 gaagatcgcc atctctaaca aggagtggct ggagtacgcc cagaccagcg tgaagcacgg
     5461 atcctatccc tatgacgtgc ccgattatgc cagcctgggc agcggctccc ccaagaaaaa
     5521 acgcaaggtg gaagatccta agaaaaagcg gaaagtggac ggcattggta gtgggagcaa
     5581 cggcagcagc ctcgatgctt tagacgattt tgacttagat atgcttggtt cagacgcgtt
     5641 agacgacttc gacctagaca tgttaggctc agatgcattg gacgacttcg atttagatat
     5701 gttgggctcc gatgccctag atgactttga tctagatatg ctaggtagtc ccaaaaagaa
     5761 gaggaaagtg ggatccgtcc ttccgcaagc gcctgcccca gcgcctgcac cggcaatggt
     5821 gtccgccctc gcacaggccc ctgcgcccgt ccccgtgctc gcgcctggac cgccccaggc
     5881 ggtcgctcca ccggctccga agccgacgca ggccggagag ggaacactct ccgaagcact
     5941 tcttcaactc cagtttgatg acgaggatct tggagcactc cttggaaact cgacagaccc
     6001 tgcggtgttt accgacctcg cgtcagtaga taactccgaa tttcagcagc ttttgaacca
     6061 gggtatcccg gtcgcgccac atacaacgga gcccatgttg atggaatacc ccgaagcaat
     6121 cacgagactt gtgacgggag cgcagcggcc tcccgatccc gcacccgcac ctttgggggc
     6181 acctggcctc cctaacggac ttttgagcgg cgacgaggat ttctcctcca tcgccgatat
     6241 ggatttctca gccttgctgt caggctctgg cagcggcagc gacctttccc atccgccccc
     6301 aaggggccat ctggatgagc tgacaaccac acttgagtcc atgaccgagg atctgaacct
     6361 ggactcaccc ctgaccccgg aattgaacga gattctggat accttcctga acgacgagtg
     6421 cctcttgcat gccatgcata tcagcacagg actgtccatc ttcgacacat ctctgtttgg
     6481 cagcagcgga tccatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     6541 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     6601 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     6661 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     6721 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     6781 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     6841 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     6901 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     6961 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     7021 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     7081 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     7141 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     7201 gggatccggc gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg
     7261 ccctatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     7321 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     7381 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     7441 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     7501 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     7561 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     7621 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     7681 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     7741 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     7801 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     7861 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     7921 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     7981 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     8041 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     8101 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     8161 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     8221 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     8281 aaaggaatag gtacaagtaa ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg
     8341 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     8401 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     8461 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     8521 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     8581 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     8641 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     8701 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     8761 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     8821 tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     8881 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     8941 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     9001 gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca
     9061 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     9121 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     9181 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     9241 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     9301 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     9361 tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     9421 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     9481 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     9541 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     9601 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     9661 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     9721 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     9781 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     9841 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     9901 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     9961 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    10021 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    10081 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    10141 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    10201 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    10261 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
    10321 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
    10381 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
    10441 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
    10501 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    10561 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    10621 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    10681 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    10741 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    10801 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    10861 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    10921 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    10981 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    11041 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    11101 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    11161 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    11221 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    11281 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    11341 tttccccgaa aagtgccacc tg
//
