LOCUS       sequence-373490-       12404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged LbCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12404)
  AUTHORS   Escobar M, Li J, Patel A, Liu S, Xu Q, Hilton IB
  TITLE     Quantification of Genome Editing and Transcriptional Control 
            Capabilities Reveals Hierarchies among Diverse CRISPR/Cas Systems in
            Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Oct 21;11(10):3239-3250. doi: 
            10.1021/acssynbio.2c00156. Epub 2022 Sep 26.
  PUBMED    36162812
REFERENCE   2  (bases 1 to 12404)
  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: 
            "10.1021/acssynbio.2c00156"; journalName: "ACS Synth Biol"; date: 
            "2022-10-21- 21"; volume: "11"; issue: "10"; pages: "3239-3250"
COMMENT     Sequence Label: LbCas12a
FEATURES             Location/Qualifiers
     source          1..12404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        2071..2146
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2208..2419
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2444..6127
                     /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             6128..6175
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6176..6199
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6209..6265
                     /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             6266..6862
                     /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     6759..6779
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6878..7466
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6931..6951)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7349..7360)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7538..7771
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7797..7814)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7803..8027
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8073..8501
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8160..8179)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8370..8391
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8510..8530)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8515..8844
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8695..8830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8757..8776
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8892..8939
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8958..9332
                     /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    9462..9583
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9499..9518)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9553..9572
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9632..9648)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9632..9648)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9645..9667)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9656..9672
                     /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(9680..9686,9687..9704,9705..9710))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9725..9746
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9863..9880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10034..10622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10114..10133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10793..11584,11585..11653))
                     /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     11416..11435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11654..11758)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11833..11852)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12024..12403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg atatcactag tgttccatta actgcgctaa aaataatttt taaatctttt
     2041 ttagcttctt gctctttttt gtacgtctct gttttagagc tagaaatagc aagttaaaat
     2101 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcgc
     2161 tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     2221 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag
     2281 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2341 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2401 gtttgccgcc agaacacagg accggttcta gagcgctgcc accatgagca agctggagaa
     2461 gtttacaaac tgctactccc tgtctaagac cctgaggttc aaggccatcc ctgtgggcaa
     2521 gacccaggag aacatcgaca ataagcggct gctggtggag gacgagaaga gagccgagga
     2581 ttataagggc gtgaagaagc tgctggatcg ctactatctg tcttttatca acgacgtgct
     2641 gcacagcatc aagctgaaga atctgaacaa ttacatcagc ctgttccgga agaaaaccag
     2701 aaccgagaag gagaataagg agctggagaa cctggagatc aatctgcgga aggagatcgc
     2761 caaggccttc aagggcaacg agggctacaa gtccctgttt aagaaggata tcatcgagac
     2821 aatcctgcca gagttcctgg acgataagga cgagatcgcc ctggtgaaca gcttcaatgg
     2881 ctttaccaca gccttcaccg gcttctttga taacagagag aatatgtttt ccgaggaggc
     2941 caagagcaca tccatcgcct tcaggtgtat caacgagaat ctgacccgct acatctctaa
     3001 tatggacatc ttcgagaagg tggacgccat ctttgataag cacgaggtgc aggagatcaa
     3061 ggagaagatc ctgaacagcg actatgatgt ggaggatttc tttgagggcg agttctttaa
     3121 ctttgtgctg acacaggagg gcatcgacgt gtataacgcc atcatcggcg gcttcgtgac
     3181 cgagagcggc gagaagatca agggcctgaa cgagtacatc aacctgtata atcagaaaac
     3241 caagcagaag ctgcctaagt ttaagccact gtataagcag gtgctgagcg atcgggagtc
     3301 tctgagcttc tacggcgagg gctatacatc cgatgaggag gtgctggagg tgtttagaaa
     3361 caccctgaac aagaacagcg agatcttcag ctccatcaag aagctggaga agctgttcaa
     3421 gaattttgac gagtactcta gcgccggcat ctttgtgaag aacggccccg ccatcagcac
     3481 aatctccaag gatatcttcg gcgagtggaa cgtgatccgg gacaagtgga atgccgagta
     3541 tgacgatatc cacctgaaga agaaggccgt ggtgaccgag aagtacgagg acgatcggag
     3601 aaagtccttc aagaagatcg gctccttttc tctggagcag ctgcaggagt acgccgacgc
     3661 cgatctgtct gtggtggaga agctgaagga gatcatcatc cagaaggtgg atgagatcta
     3721 caaggtgtat ggctcctctg agaagctgtt cgacgccgat tttgtgctgg agaagagcct
     3781 gaagaagaac gacgccgtgg tggccatcat gaaggacctg ctggattctg tgaagagctt
     3841 cgagaattac atcaaggcct tctttggcga gggcaaggag acaaacaggg acgagtcctt
     3901 ctatggcgat tttgtgctgg cctacgacat cctgctgaag gtggaccaca tctacgatgc
     3961 catccgcaat tatgtgaccc agaagcccta ctctaaggat aagttcaagc tgtattttca
     4021 gaaccctcag ttcatgggcg gctgggacaa ggataaggag acagactatc gggccaccat
     4081 cctgagatac ggctccaagt actatctggc catcatggat aagaagtacg ccaagtgcct
     4141 gcagaagatc gacaaggacg atgtgaacgg caattacgag aagatcaact ataagctgct
     4201 gcccggccct aataagatgc tgccaaaggt gttcttttct aagaagtgga tggcctacta
     4261 taaccccagc gaggacatcc agaagatcta caagaatggc acattcaaga agggcgatat
     4321 gtttaacctg aatgactgtc acaagctgat cgacttcttt aaggatagca tctcccggta
     4381 tccaaagtgg tccaatgcct acgatttcaa cttttctgag acagagaagt ataaggacat
     4441 cgccggcttt tacagagagg tggaggagca gggctataag gtgagcttcg agtctgccag
     4501 caagaaggag gtggataagc tggtggagga gggcaagctg tatatgttcc agatctataa
     4561 caaggacttt tccgataagt ctcacggcac acccaatctg cacaccatgt acttcaagct
     4621 gctgtttgac gagaacaatc acggacagat caggctgagc ggaggagcag agctgttcat
     4681 gaggcgcgcc tccctgaaga aggaggagct ggtggtgcac ccagccaact cccctatcgc
     4741 caacaagaat ccagataatc ccaagaaaac cacaaccctg tcctacgacg tgtataagga
     4801 taagaggttt tctgaggacc agtacgagct gcacatccca atcgccatca ataagtgccc
     4861 caagaacatc ttcaagatca atacagaggt gcgcgtgctg ctgaagcacg acgataaccc
     4921 ctatgtgatc ggcatcgata ggggcgagcg caatctgctg tatatcgtgg tggtggacgg
     4981 caagggcaac atcgtggagc agtattccct gaacgagatc atcaacaact tcaacggcat
     5041 caggatcaag acagattacc actctctgct ggacaagaag gagaaggaga ggttcgaggc
     5101 ccgccagaac tggacctcca tcgagaatat caaggagctg aaggccggct atatctctca
     5161 ggtggtgcac aagatctgcg agctggtgga gaagtacgat gccgtgatcg ccctggagga
     5221 cctgaactct ggctttaaga atagccgcgt gaaggtggag aagcaggtgt atcagaagtt
     5281 cgagaagatg ctgatcgata agctgaacta catggtggac aagaagtcta atccttgtgc
     5341 aacaggcggc gccctgaagg gctatcagat caccaataag ttcgagagct ttaagtccat
     5401 gtctacccag aacggcttca tcttttacat ccctgcctgg ctgacatcca agatcgatcc
     5461 atctaccggc tttgtgaacc tgctgaaaac caagtatacc agcatcgccg attccaagaa
     5521 gttcatcagc tcctttgaca ggatcatgta cgtgcccgag gaggatctgt tcgagtttgc
     5581 cctggactat aagaacttct ctcgcacaga cgccgattac atcaagaagt ggaagctgta
     5641 ctcctacggc aaccggatca gaatcttccg gaatcctaag aagaacaacg tgttcgactg
     5701 ggaggaggtg tgcctgacca gcgcctataa ggagctgttc aacaagtacg gcatcaatta
     5761 tcagcagggc gatatcagag ccctgctgtg cgagcagtcc gacaaggcct tctactctag
     5821 ctttatggcc ctgatgagcc tgatgctgca gatgcggaac agcatcacag gccgcaccga
     5881 cgtggatttt ctgatcagcc ctgtgaagaa ctccgacggc atcttctacg atagccggaa
     5941 ctatgaggcc caggagaatg ccatcctgcc aaagaacgcc gacgccaatg gcgcctataa
     6001 catcgccaga aaggtgctgt gggccatcgg ccagttcaag aaggccgagg acgagaagct
     6061 ggataaggtg aagatcgcca tctctaacaa ggagtggctg gagtacgccc agaccagcgt
     6121 gaagcacaag cgacctgccg ccacaaagaa ggctggacag gctaagaaga agaaagatta
     6181 caaagacgat gacgataagg gatccggcgc aacaaacttc tctctgctga aacaagccgg
     6241 agatgtcgaa gagaatcctg gaccgaccga gtacaagccc acggtgcgcc tcgccacccg
     6301 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     6361 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     6421 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     6481 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     6541 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     6601 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga
     6661 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     6721 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     6781 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     6841 gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat
     6901 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6961 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     7021 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     7081 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     7141 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     7201 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     7261 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     7321 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     7381 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     7441 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     7501 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     7561 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     7621 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     7681 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     7741 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
     7801 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     7861 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     7921 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     7981 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     8041 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     8101 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     8161 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     8221 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     8281 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     8341 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     8401 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     8461 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     8521 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     8581 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     8641 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     8701 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     8761 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     8821 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     8881 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     8941 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     9001 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     9061 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     9121 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     9181 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     9241 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     9301 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
     9361 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     9421 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     9481 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     9541 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     9601 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     9661 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     9721 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     9781 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     9841 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     9901 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     9961 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    10021 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    10081 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    10141 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    10201 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    10261 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    10321 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    10381 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    10441 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    10501 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    10561 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    10621 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    10681 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    10741 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    10801 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    10861 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    10921 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    10981 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    11041 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    11101 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    11161 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    11221 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    11281 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    11341 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    11401 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    11461 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    11521 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    11581 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    11641 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    11701 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    11761 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
    11821 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
    11881 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
    11941 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
    12001 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
    12061 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
    12121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
    12181 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
    12241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
    12301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
    12361 tacttggcag tacatctacg tattagtcat cgctattacc atgg
//
