LOCUS       sequence-242363-       12417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector to constitutively express humanized LbCpf1/Cas12a,
            with blasticidin resistance for selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12417)
  AUTHORS   Chow RD, Wang G, Ye L, Codina A, Kim HR, Shen L, Dong MB, Errami Y, 
            Chen S
  TITLE     In vivo profiling of metastatic double knockouts through CRISPR-Cpf1
            screens.
  JOURNAL   Nat Methods. 2019 May;16(5):405-408. doi: 10.1038/s41592-019-0371-5.
            Epub 2019 Apr 8.
  PUBMED    30962622
REFERENCE   2  (bases 1 to 12417)
  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/s41592-019-0371-5"; journalName: "Nat Methods"; date: 
            "2019-05"; volume: "16"; issue: "5"; pages: "405-408"
COMMENT     Sequence Label: pRC10-EFS-huLbCpf1-2A-Blast-WPRE
FEATURES             Location/Qualifiers
     source          1..12417
                     /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        2627..2838
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2864..6547
                     /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
                     NTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             6548..6595
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6596..6619
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6629..6685
                     /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             6686..7084
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7109..7697
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7162..7182)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7580..7591)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7700..7716)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7701..7717)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8222..8402
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8428..8445)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8434..8658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8704..9132
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8791..8810)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9001..9022
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9141..9161)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9146..9475
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9326..9461
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9388..9407
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9523..9570
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9589..9963
                     /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    10093..10214
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10130..10149)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10184..10203
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10263..10279)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10263..10279)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10276..10298)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10287..10303
                     /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(10311..10317,10318..10335,10336..10341))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10356..10377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10494..10511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10665..11253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10745..10764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11424..12215,12216..12284))
                     /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     12047..12066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12285..12389)
                     /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 atcgagtggc tccggtgccc gtcagtgggc agagcgcaca
     2641 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     2701 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     2761 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     2821 tttgccgcca gaacacaggt gtcgtgacgc gggatccgcc accatgagca agctggagaa
     2881 gtttacaaac tgctactccc tgtctaagac cctgaggttc aaggccatcc ctgtgggcaa
     2941 gacccaggag aacatcgaca ataagcggct gctggtggag gacgagaaga gagccgagga
     3001 ttataagggc gtgaagaagc tgctggatcg ctactatctg tcttttatca acgacgtgct
     3061 gcacagcatc aagctgaaga atctgaacaa ttacatcagc ctgttccgga agaaaaccag
     3121 aaccgagaag gagaataagg agctggagaa cctggagatc aatctgcgga aggagatcgc
     3181 caaggccttc aagggcaacg agggctacaa gtccctgttt aagaaggata tcatcgagac
     3241 aatcctgcca gagttcctgg acgataagga cgagatcgcc ctggtgaaca gcttcaatgg
     3301 ctttaccaca gccttcaccg gcttctttga taacagagag aatatgtttt ccgaggaggc
     3361 caagagcaca tccatcgcct tcaggtgtat caacgagaat ctgacccgct acatctctaa
     3421 tatggacatc ttcgagaagg tggacgccat ctttgataag cacgaggtgc aggagatcaa
     3481 ggagaagatc ctgaacagcg actatgatgt ggaggatttc tttgagggcg agttctttaa
     3541 ctttgtgctg acacaggagg gcatcgacgt gtataacgcc atcatcggcg gcttcgtgac
     3601 cgagagcggc gagaagatca agggcctgaa cgagtacatc aacctgtata atcagaaaac
     3661 caagcagaag ctgcctaagt ttaagccact gtataagcag gtgctgagcg atcgggagtc
     3721 tctgagcttc tacggcgagg gctatacatc cgatgaggag gtgctggagg tgtttagaaa
     3781 caccctgaac aagaacagcg agatcttcag ctccatcaag aagctggaga agctgttcaa
     3841 gaattttgac gagtactcta gcgccggcat ctttgtgaag aacggccccg ccatcagcac
     3901 aatctccaag gatatcttcg gcgagtggaa cgtgatccgg gacaagtgga atgccgagta
     3961 tgacgatatc cacctgaaga agaaggccgt ggtgaccgag aagtacgagg acgatcggag
     4021 aaagtccttc aagaagatcg gctccttttc tctggagcag ctgcaggagt acgccgacgc
     4081 cgatctgtct gtggtggaga agctgaagga gatcatcatc cagaaggtgg atgagatcta
     4141 caaggtgtat ggctcctctg agaagctgtt cgacgccgat tttgtgctgg agaagagcct
     4201 gaagaagaac gacgccgtgg tggccatcat gaaggacctg ctggattctg tgaagagctt
     4261 cgagaattac atcaaggcct tctttggcga gggcaaggag acaaacaggg acgagtcctt
     4321 ctatggcgat tttgtgctgg cctacgacat cctgctgaag gtggaccaca tctacgatgc
     4381 catccgcaat tatgtgaccc agaagcccta ctctaaggat aagttcaagc tgtattttca
     4441 gaaccctcag ttcatgggcg gctgggacaa ggataaggag acagactatc gggccaccat
     4501 cctgagatac ggctccaagt actatctggc catcatggat aagaagtacg ccaagtgcct
     4561 gcagaagatc gacaaggacg atgtgaacgg caattacgag aagatcaact ataagctgct
     4621 gcccggccct aataagatgc tgccaaaggt gttcttttct aagaagtgga tggcctacta
     4681 taaccccagc gaggacatcc agaagatcta caagaatggc acattcaaga agggcgatat
     4741 gtttaacctg aatgactgtc acaagctgat cgacttcttt aaggatagca tctcccggta
     4801 tccaaagtgg tccaatgcct acgatttcaa cttttctgag acagagaagt ataaggacat
     4861 cgccggcttt tacagagagg tggaggagca gggctataag gtgagcttcg agtctgccag
     4921 caagaaggag gtggataagc tggtggagga gggcaagctg tatatgttcc agatctataa
     4981 caaggacttt tccgataagt ctcacggcac acccaatctg cacaccatgt acttcaagct
     5041 gctgtttgac gagaacaatc acggacagat caggctgagc ggaggagcag agctgttcat
     5101 gaggcgcgcc tccctgaaga aggaggagct ggtggtgcac ccagccaact cccctatcgc
     5161 caacaagaat ccagataatc ccaagaaaac cacaaccctg tcctacgacg tgtataagga
     5221 taagaggttt tctgaggacc agtacgagct gcacatccca atcgccatca ataagtgccc
     5281 caagaacatc ttcaagatca atacagaggt gcgcgtgctg ctgaagcacg acgataaccc
     5341 ctatgtgatc ggcatcgata ggggcgagcg caatctgctg tatatcgtgg tggtggacgg
     5401 caagggcaac atcgtggagc agtattccct gaacgagatc atcaacaact tcaacggcat
     5461 caggatcaag acagattacc actctctgct ggacaagaag gagaaggaga ggttcgaggc
     5521 ccgccagaac tggacctcca tcgagaatat caaggagctg aaggccggct atatctctca
     5581 ggtggtgcac aagatctgcg agctggtgga gaagtacgat gccgtgatcg ccctggagga
     5641 cctgaactct ggctttaaga atagccgcgt gaaggtggag aagcaggtgt atcagaagtt
     5701 cgagaagatg ctgatcgata agctgaacta catggtggac aagaagtcta atccttgtgc
     5761 aacaggcggc gccctgaagg gctatcagat caccaataag ttcgagagct ttaagtccat
     5821 gtctacccag aacggcttca tcttttacat ccctgcctgg ctgacatcca agatcgatcc
     5881 atctaccggc tttgtgaacc tgctgaaaac caagtatacc agcatcgccg attccaagaa
     5941 gttcatcagc tcctttgaca ggatcatgta cgtgcccgag gaggatctgt tcgagtttgc
     6001 cctggactat aagaacttct ctcgcacaga cgccgattac atcaagaagt ggaagctgta
     6061 ctcctacggc aaccggatca gaatcttccg gaatcctaag aagaacaacg tgttcgactg
     6121 ggaggaggtg tgcctgacca gcgcctataa ggagctgttc aacaagtacg gcatcaatta
     6181 tcagcagggc gatatcagag ccctgctgtg cgagcagtcc gacaaggcct tctactctag
     6241 ctttatggcc ctgatgagcc tgatgctgca gatgcggaac agcatcacag gccgcaccga
     6301 cgtggatttt ctgatcagcc ctgtgaagaa ctccgacggc atcttctacg atagccggaa
     6361 ctatgaggcc caggagaatg ccatcctgcc aaagaacgcc gacgccaatg gcgcctataa
     6421 catcgccaga aaggtgctgt gggccatcgg ccagttcaag aaggccgagg acgagaagct
     6481 ggataaggtg aagatcgcca tctctaacaa ggagtggctg gagtacgccc agaccagcgt
     6541 gaagcacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaaggatta
     6601 caaagacgat gacgataagg gcagcggcgc caccaacttc agcctgctga agcaggccgg
     6661 cgacgtggag gagaaccccg gccccatggc caagcctttg tctcaagaag aatccaccct
     6721 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     6781 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     6841 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     6901 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     6961 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     7021 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     7081 ctaagaattc gatatcaagc ttatcggtaa tcaacctctg gattacaaaa tttgtgaaag
     7141 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     7201 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     7261 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     7321 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     7381 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     7441 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     7501 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     7561 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     7621 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     7681 ttgggccgcc tccccgcatc gataccgtcg acctcgagac ctagaaaaac atggagcaat
     7741 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     7801 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     7861 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     7921 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     7981 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     8041 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     8101 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     8161 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     8221 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     8281 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     8341 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     8401 cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     8461 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     8521 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     8581 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     8641 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     8701 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     8761 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     8821 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     8881 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     8941 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     9001 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     9061 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     9121 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     9181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     9241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     9301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     9361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     9421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     9481 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     9541 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
     9601 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     9661 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     9721 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     9781 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     9841 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     9901 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     9961 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
    10021 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
    10081 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    10141 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    10201 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
    10261 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
    10321 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
    10381 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
    10441 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
    10501 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
    10561 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
    10621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
    10681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
    10741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
    10801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
    10861 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
    10921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
    10981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
    11041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
    11101 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
    11161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
    11221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    11281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    11341 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
    11401 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    11461 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    11521 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    11581 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    11641 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    11701 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    11761 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    11821 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    11881 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    11941 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    12001 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    12061 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    12121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    12181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    12241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    12301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    12361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgac
//
