LOCUS       sequence-431402-       13915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  All-in-one, knockout, delivers Cas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13915)
  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 13915)
  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: pRDB_054
FEATURES             Location/Qualifiers
     source          1..13915
                     /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        complement(2695..2943)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2753..2772)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2923..2943)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        3017..4195
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3248..4186
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4143..4163
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4200..4209
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4245..4265
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4272..8189
                     /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      7909..7918
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8196..8243
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8250..8330
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             8337..8393
                     /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"
     CDS             8394..8987
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     8887..8907
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9007..9595
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9060..9080)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9478..9489)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9667..9900
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9926..9943)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9932..10156
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10202..10630
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10289..10308)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10499..10520
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10639..10659)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10644..10973
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10824..10959
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10886..10905
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11021..11068
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11087..11461
                     /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    11591..11712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11628..11647)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11682..11701
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11761..11777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11761..11777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11774..11796)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11785..11801
                     /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(11809..11815,11816..11833,11834..11839))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11854..11875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11992..12009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12163..12751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12243..12262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12922..13713,13714..13782))
                     /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     13545..13564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13783..13887)
                     /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 cagaaccgac tcggtgccac tttttcaaga
     2641 ccagattcag agacgagaaa aaacgcgtct ccatctacaa gagtagaaat taccggtgtt
     2701 tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag
     2761 catatgatag tccattttaa aacataattt taaaactgca aactacccaa gaaattatta
     2821 ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattctaat
     2881 tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc
     2941 ctcttcgaat ctagttgcaa agatggataa agttttaaac agagaggaat ctttgcagct
     3001 aatggacctt ctacacggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     3061 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
     3121 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     3181 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     3241 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
     3301 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
     3361 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
     3421 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
     3481 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
     3541 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
     3601 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
     3661 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
     3721 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
     3781 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
     3841 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
     3901 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
     3961 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
     4021 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
     4081 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
     4141 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaaggag
     4201 ccaccatggc caaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga
     4261 aagtcggcag cacacagttc gagggcttta ccaacctgta tcaggtgagc aagacactgc
     4321 ggtttgagct gatcccacag ggcaagaccc tgaagcacat ccaggagcag ggcttcatcg
     4381 aggaggacaa ggcccgcaat gatcactaca aggagctgaa gcccatcatc gatcggatct
     4441 acaagaccta tgccgaccag tgcctgcagc tggtgcagct ggattgggag aacctgagcg
     4501 ccgccatcga ctcctataga aaggagaaaa ccgaggagac aaggaacgcc ctgatcgagg
     4561 agcaggccac atatcgcaat gccatccacg actacttcat cggccggaca gacaacctga
     4621 ccgatgccat caataagaga cacgccgaga tctacaaggg cctgttcaag gccgagctgt
     4681 ttaatggcaa ggtgctgaag cagctgggca ccgtgaccac aaccgagcac gagaacgccc
     4741 tgctgcggag cttcgacaag tttacaacct acttctccgg cttttatcgg aacaggaaga
     4801 acgtgttcag cgccgaggat atcagcacag ccatcccaca ccgcatcgtg caggacaact
     4861 tccccaagtt taaggagaat tgtcacatct tcacacgcct gatcaccgcc gtgcccagcc
     4921 tgcgggagca ctttgagaac gtgaagaagg ccatcggcat cttcgtgagc acctccatcg
     4981 aggaggtgtt ttccttccct ttttataacc agctgctgac acagacccag atcgacctgt
     5041 ataaccagct gctgggagga atctctcggg aggcaggcac cgagaagatc aagggcctga
     5101 acgaggtgct gaatctggcc atccagaaga atgatgagac agcccacatc atcgcctccc
     5161 tgccacacag attcatcccc ctgtttaagc agatcctgtc cgataggaac accctgtctt
     5221 tcatcctgga ggagtttaag agcgacgagg aagtgatcca gtccttctgc aagtacaaga
     5281 cactgctgag aaacgagaac gtgctggaga cagccgaggc cctgtttaac gagctgaaca
     5341 gcatcgacct gacacacatc ttcatcagcc acaagaagct ggagacaatc agcagcgccc
     5401 tgtgcgacca ctgggataca ctgaggaatg ccctgtatga gcggagaatc tccgagctga
     5461 caggcaagat caccaagtct gccaaggaga aggtgcagcg cagcctgaag cacgaggata
     5521 tcaacctgca ggagatcatc tctgccgcag gcaaggagct gagcgaggcc ttcaagcaga
     5581 aaaccagcga gatcctgtcc cacgcacacg ccgccctgga tcagccactg cctacaaccc
     5641 tgaagaagca ggaggagaag gagatcctga agtctcagct ggacagcctg ctgggcctgt
     5701 accacctgct ggactggttt gccgtggatg agtccaacga ggtggacccc gagttctctg
     5761 cccggctgac cggcatcaag ctggagatgg agccttctct gagcttctac aacaaggcca
     5821 gaaattatgc caccaagaag ccctactccg tggagaagtt caagctgaac tttcagatgc
     5881 ctacactggc ccgcggctgg gacgtgaatc gggagaagaa caatggcgcc atcctgtttg
     5941 tgaagaacgg cctgtactat ctgggcatca tgccaaagca gaagggcagg tataaggccc
     6001 tgagcttcga gcccacagag aaaaccagcg agggctttga taagatgtac tatgactact
     6061 tccctgatgc cgccaagatg atcccaaagt gcagcaccca gctgaaggcc gtgacagccc
     6121 actttcagac ccacacaacc cccatcctgc tgtccaacaa tttcatcgag cctctggaga
     6181 tcacaaagga gatctacgac ctgaacaatc ctgagaagga gccaaagaag tttcagacag
     6241 cctacgccaa gaaaaccggc gaccagaagg gctacagaga ggccctgtgc aagtggatcg
     6301 acttcacaag ggattttctg tccaagtata ccaagacaac ctctatcgat ctgtctagcc
     6361 tgcggccatc ctctcagtat aaggacctgg gcgagtacta tgccgagctg aatcccctgc
     6421 tgtaccacat cagcttccag agaatcgccg agaaggagat catggatgcc gtggagacag
     6481 gcaagctgta cctgttccag atctataaca aggactttgc caagggccac cacggcaagc
     6541 ctaatctgca cacactgtat tggaccggcc tgttttctcc agagaacctg gccaagacaa
     6601 gcatcaagct gaatggccag gccgagctgt tctaccgccc taagtccagg atgaagagga
     6661 tggcacaccg gctgggagag aagatgctga acaagaagct gaaggatcag aaaaccccaa
     6721 tccccgacac cctgtaccag gagctgtacg actatgtgaa tcacagactg tcccacgacc
     6781 tgtctgatga ggccagggcc ctgctgccca acgtgatcac caaggaggtg tctcacgaga
     6841 tcatcaagga taggcgcttt accagcgaca agttcttttt ccacgtgcct atcacactga
     6901 actatcaggc cgccaattcc ccatctaagt tcaaccagag ggtgaatgcc tacctgaagg
     6961 agcaccccga gacacctatc atcggcatcg atcggggcga gagaaacctg atctatatca
     7021 cagtgatcga ctccaccggc aagatcctgg agcagcggag cctgaacacc atccagcagt
     7081 ttgattacca gaagaagctg gacaacaggg agaaggagag ggtggcagca aggcaggcct
     7141 ggtctgtggt gggcacaatc aaggatctga agcagggcta tctgagccag gtcatccacg
     7201 agatcgtgga cctgatgatc cactaccagg ccgtggtggt gctggagaac ctgaatttcg
     7261 gctttaagag caagaggacc ggcatcgccg agaaggccgt gtaccagcag ttcgagaaga
     7321 tgctgatcga taagctgaat tgcctggtgc tgaaggacta tccagcagag aaagtgggag
     7381 gcgtgctgaa cccataccag ctgacagacc agttcacctc ctttgccaag atgggcaccc
     7441 agtctggctt cctgttttac gtgcctgccc catatacatc taagatcgat cccctgaccg
     7501 gcttcgtgga ccccttcgtg tggaaaacca tcaagaatca cgagagccgc aagcacttcc
     7561 tggagggctt cgactttctg cactacgacg tgaaaaccgg cgacttcatc ctgcacttta
     7621 agatgaacag aaatctgtcc ttccagaggg gcctgcccgg ctttatgcct gcatgggata
     7681 tcgtgttcga gaagaacgag acacagtttg acgccaaggg cacccctttc atcgccggca
     7741 agagaatcgt gccagtgatc gagaatcaca gattcaccgg cagataccgg gacctgtatc
     7801 ctgccaacga gctgatcgcc ctgctggagg agaagggcat cgtgttcagg gatggctcca
     7861 acatcctgcc aaagctgctg gagaatgacg attctcacgc catcgacacc atggtggccc
     7921 tgatccgcag cgtgctgcag atgcggaact ccaatgccgc cacaggcgag gactatatca
     7981 acagccccgt gcgcgatctg aatggcgtgt gcttcgactc ccggtttcag aacccagagt
     8041 ggcccatgga cgccgatgcc aatggcgcct accacatcgc cctgaagggc cagctgctgc
     8101 tgaatcacct gaaggagagc aaggatctga agctgcagaa cggcatctcc aatcaggact
     8161 ggctggccta catccaggag ctgcgcaacg ggtccaaaag gccggcggcc acgaaaaagg
     8221 ccggccaggc aaaaaagaaa aagggatcct acccatacga tgttccagat tacgcttatc
     8281 cctacgacgt gcctgattat gcatacccat atgatgtccc cgactatgcc ggaagcgcaa
     8341 caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgaccgagt
     8401 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     8461 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     8521 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     8581 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     8641 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     8701 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     8761 tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     8821 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     8881 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     8941 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgccgga tcctgataaa
     9001 ttcgaaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     9061 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     9121 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     9181 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     9241 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     9301 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     9361 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     9421 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     9481 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     9541 cgtcctcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga
     9601 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     9661 ggggactgga agggctaatt cactcccaac gaagataaga tctgcttttt gcttgtactg
     9721 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     9781 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     9841 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     9901 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     9961 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
    10021 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
    10081 ggggtggggc aggacagcaa gggggaggat tgggaagata atagcaggca tgctggggat
    10141 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
    10201 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
    10261 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
    10321 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
    10381 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
    10441 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
    10501 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
    10561 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
    10621 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
    10681 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
    10741 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
    10801 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
    10861 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
    10921 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
    10981 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
    11041 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
    11101 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
    11161 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
    11221 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
    11281 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
    11341 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
    11401 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
    11461 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
    11521 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
    11581 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
    11641 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
    11701 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
    11761 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
    11821 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
    11881 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
    11941 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
    12001 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
    12061 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
    12121 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
    12181 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
    12241 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
    12301 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
    12361 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
    12421 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
    12481 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
    12541 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
    12601 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
    12661 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
    12721 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
    12781 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
    12841 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
    12901 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
    12961 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
    13021 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
    13081 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
    13141 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
    13201 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
    13261 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
    13321 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
    13381 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
    13441 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
    13501 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
    13561 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
    13621 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
    13681 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
    13741 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
    13801 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
    13861 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgac
//
