LOCUS       sequence-431399-       13337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knockout, EF1a-driven Cas12a (Cas only).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13337)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 13337)
  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: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: pMV_AA090
FEATURES             Location/Qualifiers
     source          1..13337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2640..3818
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2871..3809
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3766..3786
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3823..3832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3868..3888
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3895..7812
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      7532..7541
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7819..7866
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7873..7953
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             7960..8013
                     /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             8020..8418
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8429..9017
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8482..8502)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8900..8911)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9089..9322
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9348..9365)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9354..9578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9624..10052
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9711..9730)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9921..9942
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10061..10081)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10066..10395
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10246..10381
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10308..10327
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10443..10490
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10509..10883
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11013..11134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11050..11069)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11104..11123
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11183..11199)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11183..11199)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11196..11218)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11207..11223
                     /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(11231..11237,11238..11255,11256..11261))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11276..11297
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11414..11431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11585..12173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11665..11684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12344..13135,13136..13204))
                     /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     12967..12986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13205..13309)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta aggtaccggg cagaggtctc gcttttttcg agacaacacg
     2641 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     2701 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     2761 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     2821 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     2881 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     2941 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
     3001 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
     3061 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
     3121 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
     3181 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
     3241 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
     3301 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
     3361 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     3421 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     3481 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     3541 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     3601 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     3661 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     3721 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     3781 tggttcaaag tttttttctt ccatttcagg tgtcgtgaag gagccaccat ggccaaacgg
     3841 acagccgacg gaagcgagtt cgagtcacca aagaagaagc ggaaagtcgg cagcacacag
     3901 ttcgagggct ttaccaacct gtatcaggtg agcaagacac tgcggtttga gctgatccca
     3961 cagggcaaga ccctgaagca catccaggag cagggcttca tcgaggagga caaggcccgc
     4021 aatgatcact acaaggagct gaagcccatc atcgatcgga tctacaagac ctatgccgac
     4081 cagtgcctgc agctggtgca gctggattgg gagaacctga gcgccgccat cgactcctat
     4141 agaaaggaga aaaccgagga gacaaggaac gccctgatcg aggagcaggc cacatatcgc
     4201 aatgccatcc acgactactt catcggccgg acagacaacc tgaccgatgc catcaataag
     4261 agacacgccg agatctacaa gggcctgttc aaggccgagc tgtttaatgg caaggtgctg
     4321 aagcagctgg gcaccgtgac cacaaccgag cacgagaacg ccctgctgcg gagcttcgac
     4381 aagtttacaa cctacttctc cggcttttat cggaacagga agaacgtgtt cagcgccgag
     4441 gatatcagca cagccatccc acaccgcatc gtgcaggaca acttccccaa gtttaaggag
     4501 aattgtcaca tcttcacacg cctgatcacc gccgtgccca gcctgcggga gcactttgag
     4561 aacgtgaaga aggccatcgg catcttcgtg agcacctcca tcgaggaggt gttttccttc
     4621 cctttttata accagctgct gacacagacc cagatcgacc tgtataacca gctgctggga
     4681 ggaatctctc gggaggcagg caccgagaag atcaagggcc tgaacgaggt gctgaatctg
     4741 gccatccaga agaatgatga gacagcccac atcatcgcct ccctgccaca cagattcatc
     4801 cccctgttta agcagatcct gtccgatagg aacaccctgt ctttcatcct ggaggagttt
     4861 aagagcgacg aggaagtgat ccagtccttc tgcaagtaca agacactgct gagaaacgag
     4921 aacgtgctgg agacagccga ggccctgttt aacgagctga acagcatcga cctgacacac
     4981 atcttcatca gccacaagaa gctggagaca atcagcagcg ccctgtgcga ccactgggat
     5041 acactgagga atgccctgta tgagcggaga atctccgagc tgacaggcaa gatcaccaag
     5101 tctgccaagg agaaggtgca gcgcagcctg aagcacgagg atatcaacct gcaggagatc
     5161 atctctgccg caggcaagga gctgagcgag gccttcaagc agaaaaccag cgagatcctg
     5221 tcccacgcac acgccgccct ggatcagcca ctgcctacaa ccctgaagaa gcaggaggag
     5281 aaggagatcc tgaagtctca gctggacagc ctgctgggcc tgtaccacct gctggactgg
     5341 tttgccgtgg atgagtccaa cgaggtggac cccgagttct ctgcccggct gaccggcatc
     5401 aagctggaga tggagccttc tctgagcttc tacaacaagg ccagaaatta tgccaccaag
     5461 aagccctact ccgtggagaa gttcaagctg aactttcaga tgcctacact ggcccgcggc
     5521 tgggacgtga atcgggagaa gaacaatggc gccatcctgt ttgtgaagaa cggcctgtac
     5581 tatctgggca tcatgccaaa gcagaagggc aggtataagg ccctgagctt cgagcccaca
     5641 gagaaaacca gcgagggctt tgataagatg tactatgact acttccctga tgccgccaag
     5701 atgatcccaa agtgcagcac ccagctgaag gccgtgacag cccactttca gacccacaca
     5761 acccccatcc tgctgtccaa caatttcatc gagcctctgg agatcacaaa ggagatctac
     5821 gacctgaaca atcctgagaa ggagccaaag aagtttcaga cagcctacgc caagaaaacc
     5881 ggcgaccaga agggctacag agaggccctg tgcaagtgga tcgacttcac aagggatttt
     5941 ctgtccaagt ataccaagac aacctctatc gatctgtcta gcctgcggcc atcctctcag
     6001 tataaggacc tgggcgagta ctatgccgag ctgaatcccc tgctgtacca catcagcttc
     6061 cagagaatcg ccgagaagga gatcatggat gccgtggaga caggcaagct gtacctgttc
     6121 cagatctata acaaggactt tgccaagggc caccacggca agcctaatct gcacacactg
     6181 tattggaccg gcctgttttc tccagagaac ctggccaaga caagcatcaa gctgaatggc
     6241 caggccgagc tgttctaccg ccctaagtcc aggatgaaga ggatggcaca ccggctggga
     6301 gagaagatgc tgaacaagaa gctgaaggat cagaaaaccc caatccccga caccctgtac
     6361 caggagctgt acgactatgt gaatcacaga ctgtcccacg acctgtctga tgaggccagg
     6421 gccctgctgc ccaacgtgat caccaaggag gtgtctcacg agatcatcaa ggataggcgc
     6481 tttaccagcg acaagttctt tttccacgtg cctatcacac tgaactatca ggccgccaat
     6541 tccccatcta agttcaacca gagggtgaat gcctacctga aggagcaccc cgagacacct
     6601 atcatcggca tcgatcgggg cgagagaaac ctgatctata tcacagtgat cgactccacc
     6661 ggcaagatcc tggagcagcg gagcctgaac accatccagc agtttgatta ccagaagaag
     6721 ctggacaaca gggagaagga gagggtggca gcaaggcagg cctggtctgt ggtgggcaca
     6781 atcaaggatc tgaagcaggg ctatctgagc caggtcatcc acgagatcgt ggacctgatg
     6841 atccactacc aggccgtggt ggtgctggag aacctgaatt tcggctttaa gagcaagagg
     6901 accggcatcg ccgagaaggc cgtgtaccag cagttcgaga agatgctgat cgataagctg
     6961 aattgcctgg tgctgaagga ctatccagca gagaaagtgg gaggcgtgct gaacccatac
     7021 cagctgacag accagttcac ctcctttgcc aagatgggca cccagtctgg cttcctgttt
     7081 tacgtgcctg ccccatatac atctaagatc gatcccctga ccggcttcgt ggaccccttc
     7141 gtgtggaaaa ccatcaagaa tcacgagagc cgcaagcact tcctggaggg cttcgacttt
     7201 ctgcactacg acgtgaaaac cggcgacttc atcctgcact ttaagatgaa cagaaatctg
     7261 tccttccaga ggggcctgcc cggctttatg cctgcatggg atatcgtgtt cgagaagaac
     7321 gagacacagt ttgacgccaa gggcacccct ttcatcgccg gcaagagaat cgtgccagtg
     7381 atcgagaatc acagattcac cggcagatac cgggacctgt atcctgccaa cgagctgatc
     7441 gccctgctgg aggagaaggg catcgtgttc agggatggct ccaacatcct gccaaagctg
     7501 ctggagaatg acgattctca cgccatcgac accatggtgg ccctgatccg cagcgtgctg
     7561 cagatgcgga actccaatgc cgccacaggc gaggactata tcaacagccc cgtgcgcgat
     7621 ctgaatggcg tgtgcttcga ctcccggttt cagaacccag agtggcccat ggacgccgat
     7681 gccaatggcg cctaccacat cgccctgaag ggccagctgc tgctgaatca cctgaaggag
     7741 agcaaggatc tgaagctgca gaacggcatc tccaatcagg actggctggc ctacatccag
     7801 gagctgcgca acgggtccaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag
     7861 aaaaagggat cctacccata cgatgttcca gattacgctt atccctacga cgtgcctgat
     7921 tatgcatacc catatgatgt ccccgactat gccggaagcg agggcagagg aagccttcta
     7981 acatgcggtg acgtggagga gaatcccggc ccttccggga tggccaagcc tttgtctcaa
     8041 gaagaatcca ccctcattga aagagcaacg gctacaatca acagcatccc catctctgag
     8101 gactacagcg tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat
     8161 gtatatcatt ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct
     8221 gcggcagctg gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg
     8281 agcccctgcg gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata
     8341 gtgaaggaca gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt
     8401 tatgtgtggg agggctaata aattcgaaaa tcaacctctg gattacaaaa tttgtgaaag
     8461 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     8521 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     8581 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     8641 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     8701 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     8761 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     8821 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     8881 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     8941 gccggctctg cggcctcttc cgcgtcctcg ccttcgccct cagacgagtc ggatctccct
     9001 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
     9061 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagataa
     9121 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     9181 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     9241 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     9301 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     9361 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     9421 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     9481 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     9541 taatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     9601 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     9661 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     9721 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     9781 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     9841 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     9901 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     9961 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
    10021 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
    10081 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    10141 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    10201 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
    10261 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
    10321 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
    10381 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
    10441 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
    10501 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
    10561 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
    10621 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
    10681 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
    10741 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
    10801 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
    10861 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
    10921 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
    10981 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
    11041 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    11101 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
    11161 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
    11221 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
    11281 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
    11341 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
    11401 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
    11461 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
    11521 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    11581 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    11641 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    11701 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    11761 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    11821 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    11881 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    11941 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    12001 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    12061 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    12121 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
    12181 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
    12241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
    12301 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
    12361 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
    12421 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
    12481 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
    12541 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    12601 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    12661 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    12721 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    12781 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    12841 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    12901 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    12961 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    13021 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    13081 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    13141 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    13201 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    13261 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    13321 gaaaagtgcc acctgac
//
