LOCUS       sequence-492876-       13286 bp    DNA     circular SYN 17-NOV-2025
DEFINITION  Express both AsCas12a and CRISPR arrays on single transcripts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13286)
  AUTHORS   Campa CC, Weisbach NR, Santinha AJ, Incarnato D, Platt RJ
  TITLE     Multiplexed genome engineering by Cas12a and CRISPR arrays encoded 
            on single transcripts.
  JOURNAL   Nat Methods. 2019 Aug 12. pii: 10.1038/s41592-019-0508-6. doi: 
            10.1038/s41592-019-0508-6.
  PUBMED    31406383
REFERENCE   2  (bases 1 to 13286)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 17, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2019 Aug 12. pii: 10.1038/s41592-019-0508-6. doi: 
            10.1038/s41592-019-0508-6."
COMMENT     Sequence Label: pCI152-pLenti-SiT-Cas12a
FEATURES             Location/Qualifiers
     source          1..13286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(53..72)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        244..623
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        625..823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             841..1021
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1068..1193
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1686..1919
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2104..2148
                     /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    2446..2563
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     polyA_signal    2613..2661
                     /note="synthetic polyadenylation signal"
     misc_feature    2680..3268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             2786..2797
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3195..3215
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3567..3647)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             complement(3654..4247)
                     /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     complement(3734..3754)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             complement(4248..4304)
                     /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             complement(4320..4367)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4368..8288)
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      4639..4648
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(8335..8354)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8336..8354)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8399..8602)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        8603..8906
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             9044..9277
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9303..9320)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9309..9533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9579..10007
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9666..9685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9876..9897
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10016..10036)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10021..10350
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10201..10336
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10263..10282
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10398..10445
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10464..10838
                     /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    10968..11089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11005..11024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11059..11078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11138..11154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11138..11154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11151..11173)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11162..11178
                     /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(11186..11192,11193..11210,11211..11216))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11231..11252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11369..11386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11540..12128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11620..11639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12299..13090,13091..13159))
                     /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     12922..12941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13160..13264)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
       61 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
      121 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
      181 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatataccc
      241 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      301 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      361 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      421 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      481 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      541 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      601 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      661 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      721 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      781 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact
      841 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
      901 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      961 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     1021 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
     1081 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
     1141 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
     1201 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
     1261 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
     1321 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
     1381 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
     1441 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
     1501 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
     1561 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
     1621 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
     1681 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
     1741 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
     1801 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1861 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1921 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1981 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     2041 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     2101 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     2161 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     2221 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     2281 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     2341 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     2401 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2461 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2521 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2581 cagcagagat ccagtttggt taattaaggt accacacaaa aaaccaacac acagatctaa
     2641 tgaaaataaa gatcttttat tacgcgtcat ggattcgagg cggggaggcg gcccaaaggg
     2701 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     2761 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     2821 cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg
     2881 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     2941 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     3001 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     3061 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     3121 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     3181 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     3241 cacaaatttt gtaatccaga ggttgattgt cgacttaacg cgatctatgt ctagaatcta
     3301 caagagtaga aattagagac gagaaatcat gaaagcttgt gaaatagctg ttgaagctgg
     3361 attagaattt gcgtctccat ctacaagagt agaaatttcg cagaccaata ccaggatctt
     3421 gccatcctat ggaactgcct cggtgagttt tgcttttttt tttttaaagc tagggaaagg
     3481 ccaaaaagca aaacctgaga aaacaaaagg ttgttttctc aggaaaagaa aaacctttac
     3541 aaccctactg acgaatcgaa ttcttaggca tagtcgggga catcatatgg gtatgcataa
     3601 tcaggcacgt cgtagggata agcgtaatct ggaacatcgt atgggtagga tccggcaccg
     3661 ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg
     3721 gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg
     3781 ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc
     3841 ttgccctggt ggtcgggcga gactccgacg gtggccagga accacgcggg ctccttgggc
     3901 cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac
     3961 tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg
     4021 gtccagaccg ccacggcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg
     4081 acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg
     4141 acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg
     4201 gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggttgg cccgggattc
     4261 tcttcgacat cccctgcttg tttcaacagg gagaagttag tggctccgct tccggatccc
     4321 tttttctttt ttgcctggcc ggcctttttc gtggccgccg gccttttgtt gcgcagctcc
     4381 tggatgtagg ccagccagtc ctgattggag atgccgttct gcagcttcag atccttgctc
     4441 tccttcaggt gattcagcag cagctggccc ttcagggcga tgtggtaggc gccattggca
     4501 tcggcgtcca tgggccactc tgggttctga aaccgggagt cgaagcacac gccattcaga
     4561 tcgcgcacgg ggctgttgat atagtcctcg cctgtggcgg cattggagtt ccgcatctgc
     4621 agcacgctgc ggatcagggc caccatggtg tcgatggcgt gagaatcgtc attctccagc
     4681 agctttggca ggatgttgga gccatccctg aacacgatgc ccttctcctc cagcagggcg
     4741 atcagctcgt tggcaggata caggtcccgg tatctgccgg tgaatctgtg attctcgatc
     4801 actggcacga ttctcttgcc ggcgatgaaa ggggtgccct tggcgtcaaa ctgtgtctcg
     4861 ttcttctcga acacgatatc ccatgcaggc ataaagccgg gcaggcccct ctggaaggac
     4921 agatttctgt tcatcttaaa gtgcaggatg aagtcgccgg ttttcacgtc gtagtgcaga
     4981 aagtcgaagc cctccaggaa gtgcttgcgg ctctcgtgat tcttgatggt tttccacacg
     5041 aaggggtcca cgaagccggt caggggatcg atcttagatg tatatggggc aggcacgtaa
     5101 aacaggaagc cagactgggt gcccatcttg gcaaaggagg tgaactggtc tgtcagctgg
     5161 tatgggttca gcacgcctcc cactttctct gctggatagt ccttcagcac caggcaattc
     5221 agcttatcga tcagcatctt ctcgaactgc tggtacacgg ccttctcggc gatgccggtc
     5281 ctcttgctct taaagccgaa attcaggttc tccagcacca ccacggcctg gtagtggatc
     5341 atcaggtcca cgatctcgtg gatgacctgg ctcagatagc cctgcttcag atccttgatt
     5401 gtgcccacca cagaccaggc ctgccttgct gccaccctct ccttctccct gttgtccagc
     5461 ttcttctggt aatcaaactg ctggatggtg ttcaggctcc gctgctccag gatcttgccg
     5521 gtggagtcga tcactgtgat atagatcagg tttctctcgc cccgatcgat gccgatgata
     5581 ggtgtctcgg ggtgctcctt caggtaggca ttcaccctct ggttgaactt agatggggaa
     5641 ttggcggcct gatagttcag tgtgataggc acgtggaaaa agaacttgtc gctggtaaag
     5701 cgcctatcct tgatgatctc gtgagacacc tccttggtga tcacgttggg cagcagggcc
     5761 ctggcctcat cagacaggtc gtgggacagt ctgtgattca catagtcgta cagctcctgg
     5821 tacagggtgt cggggattgg ggttttctga tccttcagct tcttgttcag catcttctct
     5881 cccagccggt gtgccatcct cttcatcctg gacttagggc ggtagaacag ctcggcctgg
     5941 ccattcagct tgatgcttgt cttggccagg ttctctggag aaaacaggcc ggtccaatac
     6001 agtgtgtgca gattaggctt gccgtggtgg cccttggcaa agtccttgtt atagatctgg
     6061 aacaggtaca gcttgcctgt ctccacggca tccatgatct ccttctcggc gattctctgg
     6121 aagctgatgt ggtacagcag gggattcagc tcggcatagt actcgcccag gtccttatac
     6181 tgagaggatg gccgcaggct agacagatcg atagaggttg tcttggtata cttggacaga
     6241 aaatcccttg tgaagtcgat ccacttgcac agggcctctc tgtagccctt ctggtcgccg
     6301 gttttcttgg cgtaggctgt ctgaaacttc tttggctcct tctcaggatt gttcaggtcg
     6361 tagatctcct ttgtgatctc cagaggctcg atgaaattgt tggacagcag gatgggggtt
     6421 gtgtgggtct gaaagtgggc tgtcacggcc ttcagctggg tgctgcactt tgggatcatc
     6481 ttggcggcat cagggaagta gtcatagtac atcttatcaa agccctcgct ggttttctct
     6541 gtgggctcga agctcagggc cttatacctg cccttctgct ttggcatgat gcccagatag
     6601 tacaggccgt tcttcacaaa caggatggcg ccattgttct tctccttatt cacgtcccag
     6661 ccagaggcca gtgtaggcat ctgaaagttc agcttgaact tctccacgga gtagggcttc
     6721 ttggtggcat aatttctggc cttgttgtag aagctcagag aaggctccat ctccagcttg
     6781 atgccggtca gccgggcaga gaactcgggg tccacctcgt tggactcatc cacggcaaac
     6841 cagtccagca ggtggtacag gcccagcagg ctgtccagct gagacttcag gatctccttc
     6901 tcctcctgct tcttcagggt tgtaggcagt ggctgatcca gggcggcgtg tgcgtgggac
     6961 aggatctcgc tggttttctg cttgaaggcc tcgctcagct ccttgcctgc ggcagagatg
     7021 atctcctgca ggttgatatc ctcgtgcttc aggctgcgct gcaccttctc cttggcagac
     7081 ttggtgatct tgcctgtcag ctcggagatt ctccgctcat acagggcatt cctcagtgta
     7141 tcccagtggt cgcacagggc gctgctgatt gtctccagct tcttgtggct gatgaagatg
     7201 tgtgtcaggt cgatgctgtt cagctcgtta aacagggcct cggctgtctc cagcacgttc
     7261 tcgtttctca gcagtgtctt gtacttgcag aaggactgga tcacttcctc gtcgctctta
     7321 aactcctcca ggatgaaaga cagggtgttc ctatcggaca ggatctgctt aaacaggggg
     7381 atgaatctgt gtggcaggga ggcgatgatg tgggctgtct catcattctt ctggatggcc
     7441 agattcagca cctcgttcag gcccttgatc ttctcggtgc ctgcctcccg agagattcct
     7501 cccagcagct ggttatacag gtcgatctgg gtctgtgtca gcagctggtt ataaaaaggg
     7561 aaggaaaaca cctcctcgat ggaggtgctc acgaagatgc cgatggcctt cttcacgttc
     7621 tcaaagtgct cccgcaggct gggcacggcg gtgatcaggc gtgtgaagat gtgacaattc
     7681 tccttaaact tggggaagtt gtcctgcacg atgcggtgtg ggatggctgt gctgatatcc
     7741 tcggcgctga acacgttctt cctgttctca taaaagccgg agaagtaggt tgtaaacttg
     7801 tcgaagctcc gcagcagggc gttctcgtgc tcggttgtgg tcacggtgcc cagctgcttc
     7861 agcaccttgc cattaaacag ctcggccttg aacaggccct tgtagatctc ggcgtgtctc
     7921 ttattgatgg catcggtcag gttgtctgtc cggccgatga agtagtcgtg gatggcattg
     7981 cgatatgtgg cctgctcctc gatcagggcg ttccttgtct cctcggtttt ctcctttcta
     8041 taggagtcga tggcggcgct caggttctcc caatccagct gcaccagctg caggcactgg
     8101 tcggcatagg tcttgtagat ccgatcgatg atgggcttca gctccttgta gtgatcattg
     8161 cgggccttgt cctcctcgat gaagccctgc tcctggatgt gcttcagggt cttgccctgt
     8221 gggatcagct caaaccgcag tgtcttgctc acctgataca ggttggtaaa gccctcgaac
     8281 tgtgtcatgg tggcggtacc aagcttaagt ttaaacgcta gccagcttgg gtctccctat
     8341 agtgagtcgt attaatttcg ataagccagt aagcagtggg ttctctagtt agccagagag
     8401 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     8461 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     8521 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     8581 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     8641 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     8701 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     8761 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     8821 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     8881 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     8941 ccgtaattga ttactattaa taactagtca cgctcgagtt taagaccaat gacttacaag
     9001 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     9061 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     9121 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     9181 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     9241 ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc
     9301 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     9361 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     9421 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     9481 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     9541 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     9601 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     9661 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     9721 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     9781 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     9841 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     9901 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     9961 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
    10021 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
    10081 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
    10141 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
    10201 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
    10261 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
    10321 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
    10381 ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat
    10441 acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg
    10501 cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact
    10561 tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg
    10621 tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg
    10681 agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg
    10741 ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg
    10801 gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt
    10861 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
    10921 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
    10981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
    11041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
    11101 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
    11161 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
    11221 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
    11281 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
    11341 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
    11401 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
    11461 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
    11521 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
    11581 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
    11641 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
    11701 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
    11761 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
    11821 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
    11881 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
    11941 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
    12001 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
    12061 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
    12121 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
    12181 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
    12241 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
    12301 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
    12361 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
    12421 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
    12481 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
    12541 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
    12601 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
    12661 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
    12721 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
    12781 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
    12841 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
    12901 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
    12961 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
    13021 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
    13081 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
    13141 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
    13201 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
    13261 cgcgcacatt tccccgaaaa gtgcca
//
