LOCUS       sequence-373576-       12722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged LbdCas12a KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12722)
  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 12722)
  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: LbdCas12a KRAB
FEATURES             Location/Qualifiers
     source          1..12722
                     /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
                     NTEVRVLLKHDDNPYVIGIARGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     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             6233..6427
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             6494..6514
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6527..6583
                     /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             6584..7180
                     /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     7077..7097
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7196..7784
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7249..7269)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7667..7678)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7856..8089
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8115..8132)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8121..8345
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8391..8819
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8478..8497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8688..8709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8828..8848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8833..9162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9013..9148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9075..9094
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9210..9257
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9276..9650
                     /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    9780..9901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9817..9836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9871..9890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9950..9966)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9950..9966)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9963..9985)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9974..9990
                     /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(9998..10004,10005..10022,10023..10028))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10043..10064
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10181..10198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10352..10940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10432..10451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11111..11902,11903..11971))
                     /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     11734..11753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11972..12076)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12151..12170)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12342..12721
                     /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 ggcatcgcca 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 gatccgatgc taagtcactg actgcctggt cccggacact
     6241 ggtgaccttc aaggatgtgt ttgtggactt caccagggag gagtggaagc tgctggacac
     6301 tgctcagcag atcctgtaca gaaatgtgat gctggagaac tataagaacc tggtttcctt
     6361 gggttatcag cttactaagc cagatgtgat cctccggttg gagaagggag aagagccctg
     6421 gctggtggag agagaaattc accaagagac ccatcctgat tcagagactg catttgaaat
     6481 caaatcatca gttccgaaaa agaaacgcaa agttgctagc tccggcgcaa caaacttctc
     6541 tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgaccgagt acaagcccac
     6601 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     6661 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     6721 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     6781 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     6841 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     6901 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     6961 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     7021 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     7081 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     7141 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag tcgacaatca
     7201 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     7261 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     7321 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     7381 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     7441 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     7501 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     7561 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     7621 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     7681 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     7741 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca
     7801 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     7861 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     7921 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     7981 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     8041 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta
     8101 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     8161 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     8221 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     8281 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     8341 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     8401 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     8461 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     8521 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     8581 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     8641 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     8701 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     8761 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     8821 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     8881 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     8941 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     9001 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     9061 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     9121 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     9181 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     9241 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
     9301 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     9361 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     9421 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     9481 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     9541 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     9601 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
     9661 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     9721 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     9781 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     9841 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     9901 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     9961 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    10021 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    10081 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    10141 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    10201 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    10261 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    10321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    10381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    10441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    10501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    10561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    10621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    10681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    10741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    10801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    10861 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    10921 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    10981 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    11041 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    11101 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    11161 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    11221 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    11281 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    11341 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    11401 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    11461 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    11521 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    11581 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    11641 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    11701 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
    11761 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    11821 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    11881 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    11941 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    12001 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    12061 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    12121 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
    12181 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
    12241 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
    12301 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
    12361 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
    12421 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
    12481 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
    12541 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
    12601 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
    12661 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
    12721 gg
//
