LOCUS       sequence-370973-       14300 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral construct encoding doxycycline-inducible enCas12a-HF (via
            TetON3G).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14300)
  AUTHORS   Hughes NW, Qu Y, Zhang J, Tang W, Pierce J, Wang C, Agrawal A, Morri
            M, Neff N, Winslow MM, Wang M, Cong L
  TITLE     Machine-learning-optimized Cas12a barcoding enables the recovery of 
            single-cell lineages and transcriptional profiles.
  JOURNAL   Mol Cell. 2022 Jun 22. pii: S1097-2765(22)00537-8. doi: 
            10.1016/j.molcel.2022.06.001.
  PUBMED    35752172
REFERENCE   2  (bases 1 to 14300)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2022 Jun 22. pii: S1097-2765(22)00537-8. doi: 
            10.1016/j.molcel.2022.06.001."
COMMENT     Sequence Label: pN251
FEATURES             Location/Qualifiers
     source          1..14300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2395..2744
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     protein_bind    2403..2421
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2439..2457
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2475..2493
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2511..2529
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2547..2565
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2583..2601
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2619..2637
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2683..2707
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2756..2765
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2768..2788
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2795..6712
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLA
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      6432..6441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6713..6760
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6761..6850
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             6857..6877
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6902..6955
                     /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             6962..7654
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGH"
     promoter        7704..7915
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      7924..7933
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7930..8325
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             8335..8388
                     /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             8389..9135
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    9151..9739
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9204..9224)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9622..9633)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9811..10044
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10070..10087)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10076..10300
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10346..10774
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10433..10452)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10643..10664
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10783..10803)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10788..11117
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10968..11103
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11030..11049
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11165..11212
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11231..11605
                     /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    11735..11856
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11772..11791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11826..11845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11905..11921)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11905..11921)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11918..11940)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11929..11945
                     /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(11953..11959,11960..11977,11978..11983))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11998..12019
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12136..12153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12307..12895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12387..12406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13066..13857,13858..13926))
                     /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     13689..13708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13927..14031)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14106..14125)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gatgaggccc tttcgtcttc actcgagttt
     2401 actccctatc agtgatagag aacgtatgaa gagtttactc cctatcagtg atagagaacg
     2461 tatgcagact ttactcccta tcagtgatag agaacgtata aggagtttac tccctatcag
     2521 tgatagagaa cgtatgacca gtttactccc tatcagtgat agagaacgta tctacagttt
     2581 actccctatc agtgatagag aacgtatatc cagtttactc cctatcagtg atagagaacg
     2641 tatgtcgagg taggcgtgta cggtgggcgc ctataaaagc agagctcgtt tagtgaaccg
     2701 tcagatcgcc tggagcaatt ccacaacact tttgtcttat acttggatca ccggtgccac
     2761 catggcccca aagaagaagc ggaaggtcgg cagcacacag ttcgagggct ttaccaacct
     2821 gtatcaggtg agcaagacac tgcggtttga gctgatccca cagggcaaga ccctgaagca
     2881 catccaggag cagggcttca tcgaggagga caaggcccgc aatgatcact acaaggagct
     2941 gaagcccatc atcgatcgga tctacaagac ctatgccgac cagtgcctgc agctggtgca
     3001 gctggattgg gagaacctga gcgccgccat cgactcctat agaaaggaga aaaccgagga
     3061 gacaaggaac gccctgatcg aggagcaggc cacatatcgc aatgccatcc acgactactt
     3121 catcggccgg acagacaacc tgaccgatgc catcaataag agacacgccg agatctacaa
     3181 gggcctgttc aaggccgagc tgtttaatgg caaggtgctg aagcagctgg gcaccgtgac
     3241 cacaaccgag cacgagaacg ccctgctgcg gagcttcgac aagtttacaa cctacttctc
     3301 cggcttttat agaaacagga agaacgtgtt cagcgccgag gatatcagca cagccatccc
     3361 acaccgcatc gtgcaggaca acttccccaa gtttaaggag aattgtcaca tcttcacacg
     3421 cctgatcacc gccgtgccca gcctgcggga gcactttgag aacgtgaaga aggccatcgg
     3481 catcttcgtg agcacctcca tcgaggaggt gttttccttc cctttttata accagctgct
     3541 gacacagacc cagatcgacc tgtataacca gctgctggga ggaatctctc gggaggcagg
     3601 caccgagaag atcaagggcc tgaacgaggt gctggccctg gccatccaga agaatgatga
     3661 gacagcccac atcatcgcct ccctgccaca cagattcatc cccctgttta agcagatcct
     3721 gtccgatagg aacaccctgt ctttcatcct ggaggagttt aagagcgacg aggaagtgat
     3781 ccagtccttc tgcaagtaca agacactgct gagaaacgag aacgtgctgg agacagccga
     3841 ggccctgttt aacgagctga acagcatcga cctgacacac atcttcatca gccacaagaa
     3901 gctggagaca atcagcagcg ccctgtgcga ccactgggat acactgagga atgccctgta
     3961 tgagcggaga atctccgagc tgacaggcaa gatcaccaag tctgccaagg agaaggtgca
     4021 gcgcagcctg aagcacgagg atatcaacct gcaggagatc atctctgccg caggcaagga
     4081 gctgagcgag gccttcaagc agaaaaccag cgagatcctg tcccacgcac acgccgccct
     4141 ggatcagcca ctgcctacaa ccctgaagaa gcaggaggag aaggagatcc tgaagtctca
     4201 gctggacagc ctgctgggcc tgtaccacct gctggactgg tttgccgtgg atgagtccaa
     4261 cgaggtggac cccgagttct ctgcccggct gaccggcatc aagctggaga tggagccttc
     4321 tctgagcttc tacaacaagg ccagaaatta tgccaccaag aagccctact ccgtggagaa
     4381 gttcaagctg aactttcaga tgcctacact ggccagaggc tgggacgtga atagagagaa
     4441 gaacaatggc gccatcctgt ttgtgaagaa cggcctgtac tatctgggca tcatgccaaa
     4501 gcagaagggc aggtataagg ccctgagctt cgagcccaca gagaaaacca gcgagggctt
     4561 tgataagatg tactatgact acttccctga tgccgccaag atgatcccaa agtgcagcac
     4621 ccagctgaag gccgtgacag cccactttca gacccacaca acccccatcc tgctgtccaa
     4681 caatttcatc gagcctctgg agatcacaaa ggagatctac gacctgaaca atcctgagaa
     4741 ggagccaaag aagtttcaga cagcctacgc caagaaaacc ggcgaccaga agggctacag
     4801 agaggccctg tgcaagtgga tcgacttcac aagggatttt ctgtccaagt ataccaagac
     4861 aacctctatc gatctgtcta gcctgcggcc atcctctcag tataaggacc tgggcgagta
     4921 ctatgccgag ctgaatcccc tgctgtacca catcagcttc cagagaatcg ccgagaagga
     4981 gatcatggat gccgtggaga caggcaagct gtacctgttc cagatctata acaaggactt
     5041 tgccaagggc caccacggca agcctaatct gcacacactg tattggaccg gcctgttttc
     5101 tccagagaac ctggccaaga caagcatcaa gctgaatggc caggccgagc tgttctaccg
     5161 ccctaagtcc aggatgaaga ggatggcaca ccggctggga gagaagatgc tgaacaagaa
     5221 gctgaaggat cagaaaaccc caatccccga caccctgtac caggagctgt acgactatgt
     5281 gaatcacaga ctgtcccacg acctgtctga tgaggccagg gccctgctgc ccaacgtgat
     5341 caccaaggag gtgtctcacg agatcatcaa ggataggcgc tttaccagcg acaagttctt
     5401 tttccacgtg cctatcacac tgaactatca ggccgccaat tccccatcta agttcaacca
     5461 gagggtgaat gcctacctga aggagcaccc cgagacacct atcatcggca tcgatcgggg
     5521 cgagagaaac ctgatctata tcacagtgat cgactccacc ggcaagatcc tggagcagcg
     5581 gagcctgaac accatccagc agtttgatta ccagaagaag ctggacaaca gggagaagga
     5641 gagggtggca gcaaggcagg cctggtctgt ggtgggcaca atcaaggatc tgaagcaggg
     5701 ctatctgagc caggtcatcc acgagatcgt ggacctgatg atccactacc aggccgtggt
     5761 ggtgctggag aacctgaatt tcggctttaa gagcaagagg accggcatcg ccgagaaggc
     5821 cgtgtaccag cagttcgaga agatgctgat cgataagctg aattgcctgg tgctgaagga
     5881 ctatccagca gagaaagtgg gaggcgtgct gaacccatac cagctgacag accagttcac
     5941 ctcctttgcc aagatgggca cccagtctgg cttcctgttt tacgtgcctg ccccatatac
     6001 atctaagatc gatcccctga ccggcttcgt ggaccccttc gtgtggaaaa ccatcaagaa
     6061 tcacgagagc cgcaagcact tcctggaggg cttcgacttt ctgcactacg acgtgaaaac
     6121 cggcgacttc atcctgcact ttaagatgaa cagaaatctg tccttccaga ggggcctgcc
     6181 cggctttatg cctgcatggg atatcgtgtt cgagaagaac gagacacagt ttgacgccaa
     6241 gggcacccct ttcatcgccg gcaagagaat cgtgccagtg atcgagaatc acagattcac
     6301 cggcagatac cgggacctgt atcctgccaa cgagctgatc gccctgctgg aggagaaggg
     6361 catcgtgttc agggatggct ccaacatcct gccaaagctg ctggagaatg acgattctca
     6421 cgccatcgac accatggtgg ccctgatccg cagcgtgctg cagatgcgga actccaatgc
     6481 cgccacaggc gaggactata tcaacagccc cgtgcgcgat ctgaatggcg tgtgcttcga
     6541 ctcccggttt cagaacccag agtggcccat ggacgccgat gccaatggcg cctaccacat
     6601 cgccctgaag ggccagctgc tgctgaatca cctgaaggag agcaaggatc tgaagctgca
     6661 gaacggcatc tccaatcagg actggctggc ctacatccag gagctgcgca acaaaaggcc
     6721 ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag tacccatacg atgttccaga
     6781 ttacgctggc tatccctacg acgtgcctga ttatgcaggc agctacccat atgatgtccc
     6841 cgactatgcc gggtccccta agaagaagag aaaggtgggt accggaagcg gaggcagtgg
     6901 agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccatctag
     6961 aatggtgagc gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt
     7021 gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
     7081 gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
     7141 taccagcttc atgtacggca gcaaaacctt catcaaccac acccagggca tccccgactt
     7201 ctttaagcag tccttccccg agggcttcac atgggagaga gtcaccacat acgaagatgg
     7261 gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt
     7321 caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg
     7381 ctgggaggcc tccaccgaga cactgtaccc cgctgacggc ggcctggaag gcagagccga
     7441 catggccctg aagctcgtgg gcgggggcca cctgatctgc aaccttaaga ccacatacag
     7501 atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg acaggagact
     7561 ggaaagaatc aaggaggccg acaaagagac atacgtcgag cagcacgagg tggctgtggc
     7621 cagatactgc gacctcccta gcaaactggg gcacaaactt aattcctaag aattcgtggg
     7681 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     7741 ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga
     7801 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     7861 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggcgta
     7921 cgggccacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg
     7981 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct
     8041 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca
     8101 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc
     8161 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt
     8221 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca
     8281 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggcggatc cggagagggc
     8341 cgaggttccc tcttgacctg tggtgatgtg gaggagaatc ccggccctat gtctagactg
     8401 gacaagagca aagtcataaa ctctgctctg gaattactca atggagtcgg tatcgaaggc
     8461 ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac
     8521 gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat
     8581 acccactcct gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag
     8641 tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc
     8701 ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc
     8761 ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc
     8821 gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat
     8881 tctatgcccc cacttctgaa acaagcaatt gagctgttcg accggcaggg agccgaacct
     8941 gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa
     9001 agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     9061 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
     9121 atgctccccg ggtaaacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa
     9181 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     9241 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     9301 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     9361 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     9421 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     9481 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     9541 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     9601 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     9661 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     9721 ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc
     9781 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     9841 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     9901 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     9961 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
    10021 agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg
    10081 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
    10141 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
    10201 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
    10261 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
    10321 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
    10381 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
    10441 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
    10501 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
    10561 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
    10621 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
    10681 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
    10741 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
    10801 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
    10861 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
    10921 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
    10981 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
    11041 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
    11101 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
    11161 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
    11221 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
    11281 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
    11341 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
    11401 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
    11461 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
    11521 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
    11581 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
    11641 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
    11701 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
    11761 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
    11821 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
    11881 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
    11941 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
    12001 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
    12061 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
    12121 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
    12181 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
    12241 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
    12301 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
    12361 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
    12421 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
    12481 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
    12541 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
    12601 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
    12661 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
    12721 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
    12781 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
    12841 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
    12901 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
    12961 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
    13021 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
    13081 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
    13141 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
    13201 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
    13261 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
    13321 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
    13381 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
    13441 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
    13501 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
    13561 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
    13621 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
    13681 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
    13741 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
    13801 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
    13861 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
    13921 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
    13981 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
    14041 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
    14101 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
    14161 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
    14221 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
    14281 gccagatata cgcgcgcgtt
//
