LOCUS       sequence-373604-       14069 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged LbdCas12a VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14069)
  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 14069)
  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 VPR
FEATURES             Location/Qualifiers
     source          1..14069
                     /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"
     protein_bind    6242..6266
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6296..6445
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             6470..6490
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6920..7276
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             7295..7864
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             7874..7930
                     /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             7931..8527
                     /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     8424..8444
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8543..9131
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8596..8616)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9014..9025)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9203..9436
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9462..9479)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9468..9692
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9738..10166
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9825..9844)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10035..10056
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10175..10195)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10180..10509
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10360..10495
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10422..10441
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10557..10604
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10623..10997
                     /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    11127..11248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11164..11183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11218..11237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11297..11313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11297..11313)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11310..11332)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11321..11337
                     /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(11345..11351,11352..11369,11370..11375))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11390..11411
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11528..11545)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11699..12287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11779..11798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12458..13249,13250..13318))
                     /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     13081..13100
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13319..13423)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13498..13517)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        13689..14068
                     /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 gatcctcgcc agggatccgt cgacttgacg cgttgatatc
     6241 aacaagtttg tacaaaaaag caggctacaa agaggccagc ggttccggac gggctgacgc
     6301 attggacgat tttgatctgg atatgctggg aagtgacgcc ctcgatgatt ttgaccttga
     6361 catgcttggt tcggatgccc ttgatgactt tgacctcgac atgctcggca gtgacgccct
     6421 tgatgatttc gacctggaca tgctgattaa ctctagaagt tccggatctc cgaaaaagaa
     6481 acgcaaagtt ggtagccagt acctgcccga caccgacgac cggcaccgga tcgaggaaaa
     6541 gcggaagcgg acctacgaga cattcaagag catcatgaag aagtccccct tcagcggccc
     6601 caccgaccct agacctccac ctagaagaat cgccgtgccc agcagatcca gcgccagcgt
     6661 gccaaaacct gccccccagc cttacccctt caccagcagc ctgagcacca tcaactacga
     6721 cgagttccct accatggtgt tccccagcgg ccagatctct caggcctctg ctctggctcc
     6781 agcccctcct caggtgctgc ctcaggctcc tgctcctgca ccagctccag ccatggtgtc
     6841 tgcactggct caggcaccag cacccgtgcc tgtgctggct cctggacctc cacaggctgt
     6901 ggctccacca gcccctaaac ctacacaggc cggcgagggc acactgtctg aagctctgct
     6961 gcagctgcag ttcgacgacg aggatctggg agccctgctg ggaaacagca ccgatcctgc
     7021 cgtgttcacc gacctggcca gcgtggacaa cagcgagttc cagcagctgc tgaaccaggg
     7081 catccctgtg gcccctcaca ccaccgagcc catgctgatg gaataccccg aggccatcac
     7141 ccggctcgtg acaggcgctc agaggcctcc tgatccagct cctgcccctc tgggagcacc
     7201 aggcctgcct aatggactgc tgtctggcga cgaggacttc agctctatcg ccgatatgga
     7261 tttctcagcc ttgctgggct ctggcagcgg cagccgggat tccagggaag ggatgttttt
     7321 gccgaagcct gaggccggct ccgctattag tgacgtgttt gagggccgcg aggtgtgcca
     7381 gccaaaacga atccggccat ttcatcctcc aggaagtcca tgggccaacc gcccactccc
     7441 cgccagcctc gcaccaacac caaccggtcc agtacatgag ccagtcgggt cactgacccc
     7501 ggcaccagtc cctcagccac tggatccagc gcccgcagtg actcccgagg ccagtcacct
     7561 gttggaggat cccgatgaag agacgagcca ggctgtcaaa gcccttcggg agatggccga
     7621 tactgtgatt ccccagaagg aagaggctgc aatctgtggc caaatggacc tttcccatcc
     7681 gcccccaagg ggccatctgg atgagctgac aaccacactt gagtccatga ccgaggatct
     7741 gaacctggac tcacccctga ccccggaatt gaacgagatt ctggatacct tcctgaacga
     7801 cgagtgcctc ttgcatgcca tgcatatcag cacaggactg tccatcttcg acacatctct
     7861 gtttggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa
     7921 tcctggaccg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     7981 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     8041 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     8101 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     8161 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     8221 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     8281 caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg
     8341 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     8401 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     8461 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     8521 tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     8581 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     8641 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     8701 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     8761 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     8821 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     8881 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     8941 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     9001 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     9061 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     9121 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     9181 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     9241 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     9301 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     9361 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     9421 tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     9481 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     9541 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     9601 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     9661 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     9721 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     9781 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     9841 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     9901 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     9961 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
    10021 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
    10081 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
    10141 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
    10201 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
    10261 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
    10321 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
    10381 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
    10441 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
    10501 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
    10561 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
    10621 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
    10681 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
    10741 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
    10801 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
    10861 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
    10921 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
    10981 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
    11041 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
    11101 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
    11161 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    11221 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
    11281 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
    11341 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
    11401 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
    11461 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
    11521 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
    11581 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
    11641 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    11701 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    11761 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    11821 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    11881 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    11941 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    12001 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    12061 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    12121 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    12181 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    12241 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    12301 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    12361 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    12421 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    12481 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    12541 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    12601 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    12661 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    12721 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    12781 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    12841 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    12901 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    12961 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    13021 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    13081 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    13141 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    13201 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    13261 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    13321 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    13381 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    13441 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
    13501 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
    13561 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
    13621 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
    13681 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
    13741 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
    13801 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
    13861 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
    13921 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
    13981 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
    14041 ctacgtatta gtcatcgcta ttaccatgg
//
