LOCUS       Exported               12502 bp ds-DNA     circular SYN 02-JAN-2023
DEFINITION  Lentiviral expression of EnAsCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12502)
  TITLE     Optimization of AsCas12a for combinatorial genetic screens in human 
            cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12502)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12502
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        32..258
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             259..439
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    486..611
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1104..1337
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1844..1959
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2111..3288
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     3236..3256
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3297..3306
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3309..3329
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3354..7271
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     CDS             7272..7319
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7326..7406
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             7422..7475
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7482..7877
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             7899..8597
                     /codon_start=1
                     /locus_tag="BFP"
                     /label=BFP
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    8607..9195
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8660..8680)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     9193..9206
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             9269..9502
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9574..9695
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9611..9630)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9665..9684
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9735..9870
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9797..9816
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(9820..9831)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(9903..9919)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9903..9919)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9916..9938)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9927..9943
                     /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(9951..9981)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9996..10017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10134..10151)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10305..10893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10385..10404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11064..11924)
                     /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     11687..11706
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11925..12029)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12097..12115
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(12153..12175)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     12275..12294
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(12488..12502,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(12502,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagctgacgc gtgtagtctt atgcaatact cttgtagtct
       61 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
      121 tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt
      181 ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc
      241 ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
      301 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
      361 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
      421 gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag
      481 aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc
      541 ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt
      601 gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa
      661 ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag
      721 aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg
      781 gacagctaca accatccctt cagacaggat cagaagaact tagatcatta tataatacag
      841 tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag
      901 acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccactgatc
      961 ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa
     1021 gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag
     1081 agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga
     1141 agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt
     1201 atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa
     1261 ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta
     1321 aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct
     1381 gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc
     1441 tggatggagt gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa
     1501 gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca
     1561 agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg
     1621 atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga
     1681 gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc
     1741 gacaggcccg aaggaataga agaagaaggt ggagagagag acagagacag atccattcga
     1801 ttagtgaacg gatctcgacg gtatcgttgc ttggatcgag caatttaaaa gaaaaggggg
     1861 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1921 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttc tgcgttgttg tcggtgctcg
     1981 ttctctgctc ttcacgctac tgaattcatc accggttctt cgaatctagt tgcaaagatg
     2041 gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg
     2101 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     2161 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
     2221 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     2281 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     2341 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt
     2401 gccttgaatt acttccactg gctgcagtac gtgattcttg atcccgagct tcgggttgga
     2461 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
     2521 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
     2581 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
     2641 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
     2701 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
     2761 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
     2821 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
     2881 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
     2941 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     3001 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     3061 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     3121 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     3181 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     3241 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgatt ctagccgcca
     3301 ccatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccacacagt
     3361 tcgagggctt taccaacctg tatcaggtga gcaagacact gcggtttgag ctgatcccac
     3421 agggcaagac cctgaagcac atccaggagc agggcttcat cgaggaggac aaggcccgca
     3481 atgatcacta caaggagctg aagcccatca tcgatcggat ctacaagacc tatgccgacc
     3541 agtgcctgca gctggtgcag ctggattggg agaacctgag cgccgccatc gactcctata
     3601 gaaaggagaa aaccgaggag acaaggaacg ccctgatcga ggagcaggcc acatatcgca
     3661 atgccatcca cgactacttc atcggccgga cagacaacct gaccgatgcc atcaataaga
     3721 gacacgccga gatctacaag ggcctgttca aggccgagct gtttaatggc aaggtgctga
     3781 agcagctggg caccgtgacc acaaccgagc acgagaacgc cctgctgcgg agcttcgaca
     3841 agtttacaac ctacttctcc ggcttttatc ggaacaggaa gaacgtgttc agcgccgagg
     3901 atatcagcac agccatccca caccgcatcg tgcaggacaa cttccccaag tttaaggaga
     3961 attgtcacat cttcacacgc ctgatcaccg ccgtgcccag cctgcgggag cactttgaga
     4021 acgtgaagaa ggccatcggc atcttcgtga gcacctccat cgaggaggtg ttttccttcc
     4081 ctttttataa ccagctgctg acacagaccc agatcgacct gtataaccag ctgctgggag
     4141 gaatctctcg ggaggcaggc accgagaaga tcaagggcct gaacgaggtg ctgaatctgg
     4201 ccatccagaa gaatgatgag acagcccaca tcatcgcctc cctgccacac agattcatcc
     4261 ccctgtttaa gcagatcctg tccgatagga acaccctgtc tttcatcctg gaggagttta
     4321 agagcgacga ggaagtgatc cagtccttct gcaagtacaa gacactgctg agaaacgaga
     4381 acgtgctgga gacagccgag gccctgttta acgagctgaa cagcatcgac ctgacacaca
     4441 tcttcatcag ccacaagaag ctggagacaa tcagcagcgc cctgtgcgac cactgggata
     4501 cactgaggaa tgccctgtat gagcggagaa tctccgagct gacaggcaag atcaccaagt
     4561 ctgccaagga gaaggtgcag cgcagcctga agcacgagga tatcaacctg caggagatca
     4621 tctctgccgc aggcaaggag ctgagcgagg ccttcaagca gaaaaccagc gagatcctgt
     4681 cccacgcaca cgccgccctg gatcagccac tgcctacaac cctgaagaag caggaggaga
     4741 aggagatcct gaagtctcag ctggacagcc tgctgggcct gtaccacctg ctggactggt
     4801 ttgccgtgga tgagtccaac gaggtggacc ccgagttctc tgcccggctg accggcatca
     4861 agctggagat ggagccttct ctgagcttct acaacaaggc cagaaattat gccaccaaga
     4921 agccctactc cgtggagaag ttcaagctga actttcagat gcctacactg gcccgcggct
     4981 gggacgtgaa tcgggagaag aacaatggcg ccatcctgtt tgtgaagaac ggcctgtact
     5041 atctgggcat catgccaaag cagaagggca ggtataaggc cctgagcttc gagcccacag
     5101 agaaaaccag cgagggcttt gataagatgt actatgacta cttccctgat gccgccaaga
     5161 tgatcccaaa gtgcagcacc cagctgaagg ccgtgacagc ccactttcag acccacacaa
     5221 cccccatcct gctgtccaac aatttcatcg agcctctgga gatcacaaag gagatctacg
     5281 acctgaacaa tcctgagaag gagccaaaga agtttcagac agcctacgcc aagaaaaccg
     5341 gcgaccagaa gggctacaga gaggccctgt gcaagtggat cgacttcaca agggattttc
     5401 tgtccaagta taccaagaca acctctatcg atctgtctag cctgcggcca tcctctcagt
     5461 ataaggacct gggcgagtac tatgccgagc tgaatcccct gctgtaccac atcagcttcc
     5521 agagaatcgc cgagaaggag atcatggatg ccgtggagac aggcaagctg tacctgttcc
     5581 agatctataa caaggacttt gccaagggcc accacggcaa gcctaatctg cacacactgt
     5641 attggaccgg cctgttttct ccagagaacc tggccaagac aagcatcaag ctgaatggcc
     5701 aggccgagct gttctaccgc cctaagtcca ggatgaagag gatggcacac cggctgggag
     5761 agaagatgct gaacaagaag ctgaaggatc agaaaacccc aatccccgac accctgtacc
     5821 aggagctgta cgactatgtg aatcacagac tgtcccacga cctgtctgat gaggccaggg
     5881 ccctgctgcc caacgtgatc accaaggagg tgtctcacga gatcatcaag gataggcgct
     5941 ttaccagcga caagttcttt ttccacgtgc ctatcacact gaactatcag gccgccaatt
     6001 ccccatctaa gttcaaccag agggtgaatg cctacctgaa ggagcacccc gagacaccta
     6061 tcatcggcat cgatcggggc gagagaaacc tgatctatat cacagtgatc gactccaccg
     6121 gcaagatcct ggagcagcgg agcctgaaca ccatccagca gtttgattac cagaagaagc
     6181 tggacaacag ggagaaggag agggtggcag caaggcaggc ctggtctgtg gtgggcacaa
     6241 tcaaggatct gaagcagggc tatctgagcc aggtcatcca cgagatcgtg gacctgatga
     6301 tccactacca ggccgtggtg gtgctggaga acctgaattt cggctttaag agcaagagga
     6361 ccggcatcgc cgagaaggcc gtgtaccagc agttcgagaa gatgctgatc gataagctga
     6421 attgcctggt gctgaaggac tatccagcag agaaagtggg aggcgtgctg aacccatacc
     6481 agctgacaga ccagttcacc tcctttgcca agatgggcac ccagtctggc ttcctgtttt
     6541 acgtgcctgc cccatataca tctaagatcg atcccctgac cggcttcgtg gaccccttcg
     6601 tgtggaaaac catcaagaat cacgagagcc gcaagcactt cctggagggc ttcgactttc
     6661 tgcactacga cgtgaaaacc ggcgacttca tcctgcactt taagatgaac agaaatctgt
     6721 ccttccagag gggcctgccc ggctttatgc ctgcatggga tatcgtgttc gagaagaacg
     6781 agacacagtt tgacgccaag ggcacccctt tcatcgccgg caagagaatc gtgccagtga
     6841 tcgagaatca cagattcacc ggcagatacc gggacctgta tcctgccaac gagctgatcg
     6901 ccctgctgga ggagaagggc atcgtgttca gggatggctc caacatcctg ccaaagctgc
     6961 tggagaatga cgattctcac gccatcgaca ccatggtggc cctgatccgc agcgtgctgc
     7021 agatgcggaa ctccaatgcc gccacaggcg aggactatat caacagcccc gtgcgcgatc
     7081 tgaatggcgt gtgcttcgac tcccggtttc agaacccaga gtggcccatg gacgccgatg
     7141 ccaatggcgc ctaccacatc gccctgaagg gccagctgct gctgaatcac ctgaaggaga
     7201 gcaaggatct gaagctgcag aacggcatct ccaatcagga ctggctggcc tacatccagg
     7261 agctgcgcaa caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg
     7321 gatcctaccc atacgatgtt ccagattacg cttatcccta cgacgtgcct gattatgcat
     7381 acccatatga tgtccccgac tatgccgcgg ccgcaggatg cgagggcaga ggaagccttc
     7441 taacatgcgg tgacgtggag gagaatcccg gcccttccgg gatggccaag cctttgtctc
     7501 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
     7561 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
     7621 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
     7681 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
     7741 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
     7801 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
     7861 gttatgtgtg ggagggcgtc gacggcagtg gaagtggcag cgagctgatt aaggagaaca
     7921 tgcacatgaa gctctacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg
     7981 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg
     8041 gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct
     8101 tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca
     8161 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca
     8221 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca
     8281 acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgaa acgctgtacc
     8341 ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc
     8401 atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga
     8461 tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga
     8521 cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg
     8581 ggcacaagct taattaatag gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     8641 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     8701 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     8761 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     8821 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     8881 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     8941 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     9001 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     9061 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     9121 cggctctgcg gcctcttccg cgtctccgcc ttcgccctca gacgagtcgg atctctcttt
     9181 gggccgcctc cccgcctggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     9241 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagataa
     9301 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     9361 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     9421 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     9481 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     9541 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     9601 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     9661 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     9721 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     9781 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     9841 agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagaccaa attcgtaatc
     9901 atgtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     9961 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
    10021 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
    10081 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
    10141 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
    10201 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
    10261 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
    10321 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
    10381 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
    10441 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
    10501 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
    10561 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
    10621 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
    10681 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
    10741 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
    10801 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
    10861 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    10921 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    10981 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
    11041 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    11101 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    11161 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    11221 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    11281 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    11341 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    11401 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    11461 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    11521 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    11581 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    11641 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    11701 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    11761 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    11821 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    11881 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    11941 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    12001 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
    12061 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
    12121 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
    12181 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
    12241 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
    12301 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
    12361 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
    12421 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
    12481 gggttttccc agtcacgacg tt
//
