LOCUS       sequence-496074-       14851 bp    DNA     circular SYN 02-JAN-2026
DEFINITION  Lentiviral expression of dCas9-KRAB with  with H2B-mCherry reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14851)
  TITLE     Brock Lab Plasmids 2025
REFERENCE   2  (bases 1 to 14851)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 2, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: lenti-EF1a-dCas9-KRAB-H2B-mCherry
FEATURES             Location/Qualifiers
     source          1..14851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    667..784
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1082..1126)
                     /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    1311..1544
                     /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    2037..2162
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2209..2389
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2407..2605)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2432..2452)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2607..2986
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3158..3177
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3252..3356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3357..3425,3426..4217)
                     /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     complement(3575..3594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4388..4976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4877..4896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5130..5147
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5264..5285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5300..5305,5306..5323,5324..5330)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5338..5354
                     /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)."
     primer_bind     5343..5365
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5362..5378
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5362..5378
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5427..5548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5438..5457)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5492..5511
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5678..6052)
                     /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"
     promoter        complement(6071..6118)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(6166..6495)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6180..6315
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6234..6253)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6480..6500
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6509..6937)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6619..6640)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6831..6850
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6983..7207
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7196..7213
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             7239..7419
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     7924..7940
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     7925..7941
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7944..8532
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             8050..8061
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     8459..8479
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8558..9268)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(8836..8855)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     9105..9123
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      9265..9274
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(9287..9664)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             complement(9677..9730)
                     /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             complement(9752..9946)
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             complement(9971..9991)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(10046..11560,11561..11563,11564..14119,
                     14120..14122,14123..14146))
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(14147..14194)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     promoter        complement(join(14210..14851,1..537))
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(14219..14851,1..306)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(14242..14262)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gaaaaaaagc gtcgcagcag gtcatcaaaa attttaaatg gctagagact tatcgaaagc
       61 agcgagacag gcgcgaaggt gccaccagat tcgcacgcgg cggccccagc gcccaggcca
      121 ggcctcaact caagcacgag gcgaaggggc tccttaagcg caaggcctcg aactctccca
      181 cccacttcca acccgaagct cgggatcaag aatcacgtac tgcagccagg tggaagtaat
      241 tcaaggcacg caagggccat aacccgtaaa gaggccaggc ccgcgggaac cacacacggc
      301 acttacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc
      361 acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact
      421 ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga cccctccccc
      481 caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac cggagccaat
      541 tcccactcct ttcaagacct agaaggtcca ttagctgcaa agattcctct ctgtttaaaa
      601 ctttatccat ctttgcagct agctaattaa ccaaactgga tctctgctgt ccctgtaata
      661 aacccgaaaa ttttgaattt ttgtaatttg tttttgtaat tctttagttt gtatgtctgt
      721 tgctattatg tctactattc tttcccctgc actgtacccc ccaatccccc cttttctttt
      781 aaaattgtgg atgaatactg ccatttgtct gcagaattgg cgcacgcagt gccgatccgt
      841 tcactaatcg aatggatctg tctctgtctc tctctccacc ttcttcttct attccttcgg
      901 gcctgtcggg tcccctcggg gttgggaggt gggtctgaaa cgataatggt gaatatccct
      961 gcctaactct attcactata gaaagtacag caaaaactat tcttaaacct accaagcctc
     1021 ctactatcat tatgaataat tttatatacc acagccaatt tgttatgtta aaccaattcc
     1081 acaaacttgc ccatttatct aattccaata attcttgttc attcttttct tgctggtttt
     1141 gcgattcttc aattaaggag tgtattaagc ttgtgtaatt gttaatttct ctgtcccact
     1201 ccatccaggt cgtgtgattc caaatctgtt ccagagattt attactccaa ctagcattcc
     1261 aaggcacagc agtggtgcaa atgagttttc cagagcaacc ccaaatcccc aggagctgtt
     1321 gatcctttag gtatctttcc acagccagga ttcttgcctg gagctgcttg atgccccaga
     1381 ctgtgagttg caacagatgc tgttgcgcct caatagccct cagcaaattg ttctgctgct
     1441 gcactatacc agacaataat tgtctggcct gtaccgtcag cgtcattgac gctgcgccca
     1501 tagtgcttcc tgctgctccc aagaacccaa ggaacaaagc tcctattccc actgctcttt
     1561 tttctctctg caccactctt ctctttgcct tggtgggtgc tactcctaat ggttcaattt
     1621 ttactacttt atatttatat aattcacttc tccaattgtc cctcatatct cctcctccag
     1681 gtctgaagat cagcggccgc ttgctgtgcg gtggtcttac ttttgttttg ctcttcctct
     1741 atcttgtcta aagcttcctt ggtgtctttt atctctatcc tttgatgcac acaatagagg
     1801 gttgctactg tattatataa tgatctaagt tcttctgatc ctgtctgaag ggatggttgt
     1861 agctgtccca gtatttgtct acagccttct gatgtttcta acaggccagg attaactgcg
     1921 aatcgttcta gctccctgct tgcccatact atatgtttta atttatattt tttctttccc
     1981 cctggcctta accgaatttt ttcccatcgc gatctaattc tcccccgctt aatactgacg
     2041 ctctcgcacc catctctctc cttctagcct ccgctagtca aaatttttgg cgtactcacc
     2101 agtcgccgcc cctcgcctct tgccgtgcgc gcttcagcaa gccgagtcct gcgtcgagag
     2161 agctcctctg gtttcccttt cgctttcaag tccctgttcg ggcgccactg ctagagattt
     2221 tccacactga ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg
     2281 cacacactac ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta
     2341 gttagccaga gagctcccag gctcagatct ggtctaacca gagagaccca gtacaggcaa
     2401 aacgcgctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     2461 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     2521 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     2581 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     2641 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     2701 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     2761 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     2821 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     2881 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     2941 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgtcaacg cgtatatctg
     3001 gcccgtacat cgcgaagcag cgcaaaacgc ctaaccctaa gcagattctt catgcaattg
     3061 tcggtcaagc cttgccttgt tgtagcttaa attttgctcg cgcactactc agcgacctcc
     3121 aacacacaag cagggagcag atactggctt aactatgcgg catcagagca gattgtactg
     3181 agagtgcacc ataggggatc gggagatctc ccgatccgtc gacgtcaggt ggcacttttc
     3241 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3301 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3361 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3421 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3481 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3541 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3601 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3661 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3721 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3781 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3841 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3901 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3961 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4021 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4081 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4141 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4201 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4261 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4321 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4381 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4441 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4501 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4561 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4621 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4681 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4741 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4801 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4861 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4921 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4981 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5041 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5101 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5161 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5221 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5281 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5341 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc
     5401 tagaggtcga cggtatacag acatgataag atacattgat gagtttggac aaaccacaac
     5461 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     5521 aaccattata agctgcaata aacaagttgg ggtgggcgaa gaactccagc atgagatccc
     5581 cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc aacctttcat
     5641 agaaggcggc ggtggaatcg aaatctcgta gcacgtgtca gtcctgctcc tcggccacga
     5701 agtgcacgca gttgccggcc gggtcgcgca gggcgaactc ccgcccccac ggctgctcgc
     5761 cgatctcggt catggccggc ccggaggcgt cccggaagtt cgtggacacg acctccgacc
     5821 actcggcgta cagctcgtcc aggccgcgca cccacaccca ggccagggtg ttgtccggca
     5881 ccacctggtc ctggaccgcg ctgatgaaca gggtcacgtc gtcccggacc acaccggcga
     5941 agtcgtcctc cacgaagtcc cgggagaacc cgagccggtc ggtccagaac tcgaccgctc
     6001 cggcgacgtc gcgcgcggtg agcaccggaa cggcactggt caacttggcc atggtttagt
     6061 tcctcacctt gtcgtattat actatgccga tatactatgc cgatgattaa ttgtcaacac
     6121 gtgctgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
     6181 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
     6241 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
     6301 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
     6361 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
     6421 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6481 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
     6541 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     6601 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     6661 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     6721 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     6781 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     6841 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     6901 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
     6961 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
     7021 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
     7081 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
     7141 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
     7201 ggcacagtcg aggctgatca gcgggtttaa acgggccctg ctagagattt tccacactga
     7261 ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac
     7321 ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga
     7381 gagctcccag gctcagatct ggtctaacca gagagaccca gtacagtccg gatgcagctc
     7441 tcgggccatg tgatgaaatg ctaggcggct gtcaaacctc cactctaata cttctctctc
     7501 cgggtcatcc atcccatgca ggctcacagg gtgtaacaag cgggtgttct ctccttcatt
     7561 ggcttcttct accttctctt gctcaactgg tactagcttg tagcaccatc caaaggtcag
     7621 tggatatctg atccctggcc ctggtgtgta gttctgccaa tcagggaagt agccttgtgt
     7681 gtggtagatc cacagatcaa ggatatcttg tcttcgttgg gagtgaatta gcccttccag
     7741 tccccccttt tcttttaaaa agtggctaag atctacagct gccttgtaag tcattggtct
     7801 taaaggtacc tgaggtgtga ctggaaaacc cacctcctcc tcctcttgtg cttctagcca
     7861 ggcacaatca gcattggtag ctgctgtatt gctacttgtg attgctccat gtttttctag
     7921 gtctcgaggt cgacggtatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg
     7981 agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag
     8041 gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt
     8101 ggcaacacag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt
     8161 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
     8221 cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac aggtggtggc
     8281 aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct
     8341 gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa
     8401 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
     8461 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
     8521 cagaggttga ttaccgataa gcttgatatc gttcgaatta cttgtacagc tcgtccatgc
     8581 cgccggtgga gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt
     8641 tgtgggaggt gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct
     8701 tcttggcctt gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca
     8761 gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg
     8821 cctcccagcc catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc
     8881 gcagcttcac cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca
     8941 ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca
     9001 gcttcaagta gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga
     9061 actgagggga caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct
     9121 tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt
     9181 ggccgttcac ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca
     9241 tgttatcctc ctcgcccttg ctcaccatgg tggcgaccgg tggatcctta gcgctggtgt
     9301 acttggtgat ggccttagta ccctcggaca cggcgtgctt ggccaactcc ccaggcagca
     9361 gcaggcgcac ggccgtctgg atctccctgg aggtgatggt cgagcgcttg ttgtaatgcg
     9421 ccaggcggga agcctcacct gcgatgcgct cgaaaatgtc gttcacaaac gaattcatga
     9481 tgcccatggc cttggacgaa atgccggtgt cagggtggac ctgcttcaga accttgtaca
     9541 catagatgga atagctctcc ttgcggctgc gcttgcgctt cttgccgcct ttcttctgcg
     9601 ccttagtcac cgccttcttg gagccctttt tcggggcggg agcagacttc gctggctctg
     9661 gcatttcgaa ggtcatgggt ccgggattct cttctacgtc tccgcatgtg agcagggaac
     9721 ctcttccctc ccctgaacct gtacagttaa ttaccagcca aggttcttcc cccttttcga
     9781 gcctaaggat cacatccggt ttcgtcaact ggtatccgag actgacgagg ttcttgtaat
     9841 tctcaagcat gacatttcga tacacaattt gttgcgcagt atccagcaat ttccactcct
     9901 cacgtgtaaa gtccacaaac acgtccttga acgtaaccag tgttctagcc cgtccggatc
     9961 cagcggccgc caccttcctc tttttcttag gtccgctacc tcctcctccg cttcctcctc
    10021 ctccgcttcc acctcctcca gcgctgtcgc ctcccagctg agacaggtcg atccgtgtct
    10081 cgtacaggcc ggtgatgctc tggtggatca gggtggcgtc cagcacctct ttggtgctgg
    10141 tgtacctctt ccggtcgatg gtggtgtcaa agtacttgaa ggcggcaggg gctcccagat
    10201 tggtcagggt aaacaggtgg atgatattct cggcctgctc tctgatgggc ttatcccggt
    10261 gcttgttgta ggcggacagc actttgtcca gattagcgtc ggccaggatc actctcttgg
    10321 agaactcgct gatctgctcg atgatctcgt ccaggtagtg cttgtgctgt tccacaaaca
    10381 gctgtttctg ctcattatcc tcgggggagc ccttcagctt ctcatagtgg ctggccaggt
    10441 acaggaagtt cacatatttg gagggcaggg ccagttcgtt tcccttctgc agttcgccgg
    10501 cagaggccag cattctcttc cggccgtttt ccagctcgaa cagggagtac ttaggcagct
    10561 tgatgatcag gtcctttttc acttctttgt agcccttggc ttccagaaag tcgatgggat
    10621 tcttctcgaa gctgcttctt tccatgatgg tgatccccag cagctctttc acactcttca
    10681 gtttcttgga cttgcccttt tccactttgg ccaccaccag cacagaatag gccacggtgg
    10741 ggctgtcgaa gccgccgtac ttcttagggt cccagtcctt ctttctggcg atcagcttat
    10801 cgctgttcct cttgggcagg atagactctt tgctgaagcc gcctgtctgc acctcggtct
    10861 ttttcacgat attcacttgg ggcatgctca gcactttccg cacggtggca aaatcccggc
    10921 ccttatccca cacgatctcc ccggtttcgc cgtttgtctc gatcagaggc cgcttccgga
    10981 tctcgccgtt ggccagggta atctcggtct tgaaaaagtt catgatgttg ctgtagaaga
    11041 agtacttggc ggtagccttg ccgatttcct gctcgctctt ggcgatcatc ttccgcacgt
    11101 cgtacacctt gtagtcgccg tacacgaact cgctttccag cttagggtac tttttgatca
    11161 gggcggttcc cacgacggcg ttcaggtagg cgtcgtgggc gtggtggtag ttgttgatct
    11221 cgcgcacttt gtaaaactgg aaatccttcc ggaaatcgga caccagcttg gacttcaggg
    11281 tgatcacttt cacttcccgg atcagcttgt cattctcgtc gtacttagtg ttcatccggg
    11341 agtccaggat ctgtgccacg tgctttgtga tctgccgggt ttccaccagc tgtctcttga
    11401 tgaagccggc cttatccagt tcgctcaggc cgcctctctc ggccttggtc agattgtcga
    11461 actttctctg ggtaatcagc ttggcgttca gcagctgccg ccagtagttc ttcatcttct
    11521 tcacgacctc ttcggagggc acgttgtcgc tcttgccccg ggccttgtcg cttctggtca
    11581 gcaccttgtt gtcgatggag tcgtccttca gaaagctctg aggcacgatg tggtccacat
    11641 cgtagtcgga cagccggttg atgtccagtt cctggtccac gtacatatcc cgcccattct
    11701 gcaggtagta caggtacagc ttctcgttct gcagctgggt gttttccacg gggtgttctt
    11761 tcaggatctg gctgcccagc tctttgatgc cctcttcgat ccgcttcatt ctctcgcggc
    11821 tgttcttctg tcccttctgg gtggtctggt tctctctggc catttcgatc acgatgttct
    11881 cgggcttgtg ccggcccatc actttcacga gctcgtccac caccttcact gtctgcagga
    11941 tgcccttctt aatggcgggg ctgccggcca gattggcaat gtgctcgtgc aggctatcgc
    12001 cctggccgga cacctgggct ttctggatgt cctctttaaa ggtcaggctg tcgtcgtgga
    12061 tcagctgcat gaagtttctg ttggcgaagc cgtcggactt caggaaatcc aggattgtct
    12121 tgccggactg cttgtcccgg atgccgttga tcagcttccg gctcagcctg ccccagccgg
    12181 tgtatctccg ccgcttcagc tgcttcatca ctttgtcgtc gaacaggtgg gcataggttt
    12241 tcagccgttc ctcgatcatc tctctgtcct caaacagtgt cagggtcagc acgatatctt
    12301 ccagaatgtc ctcgttttcc tcattgtcca ggaagtcctt gtccttgata attttcagca
    12361 gatcgtggta tgtgcccagg gaggcgttga accgatcttc cacgccggag atttccacgg
    12421 agtcgaagca ctcgattttc ttgaagtagt cctctttcag ctgcttcacg gtcactttcc
    12481 ggttggtctt gaacagcagg tccacgatgg cctttttctg ctcgccgctc aggaaggcgg
    12541 gctttctcat tccctcggtc acgtatttca ctttggtcag ctcgttatac acggtgaagt
    12601 actcgtacag caggctgtgc ttgggcagca ccttctcgtt gggcaggttc ttatcgaagt
    12661 tggtcatccg ctcgatgaag ctctgggcgg aagcgccctt gtccaccact tcctcgaagt
    12721 tccagggggt gatggtttcc tcgctctttc tggtcatcca ggcgaatctg ctgtttcccc
    12781 tggccagagg gcccacgtag taggggatgc ggaaggtcag gatcttctcg atcttttccc
    12841 ggttgtcctt caggaatggg taaaaatctt cctgccgccg cagaatggcg tgcagctctc
    12901 ccaggtggat ctggtggggg atgctgccgt tgtcgaaggt ccgctgcttc cgcagcaggt
    12961 cctctctgtt cagcttcacg agcagttcct cggtgccgtc catcttttcc aggatgggct
    13021 tgatgaactt gtagaactct tcctggctgg ctccgccgtc aatgtagccg gcgtagccgt
    13081 tcttgctctg gtcgaagaaa atctctttgt acttctcagg cagctgctgc cgcacgagag
    13141 ctttcagcag ggtcaggtcc tggtggtgct cgtcgtatct cttgatcata gaggcgctca
    13201 ggggggcctt ggtgatctcg gtgttcactc tcaggatgtc gctcagcagg atggcgtcgg
    13261 acaggttctt ggcggccaga aacaggtcgg cgtactggtc gccgatctgg gccagcaggt
    13321 tgtccaggtc gtcgtcgtag gtgtccttgc tcagctgcag tttggcatcc tcggccaggt
    13381 cgaagttgct cttgaagttg ggggtcaggc ccaggctcag ggcaatcagg ttgccgaaca
    13441 ggccattctt cttctcgccg ggcagctggg cgatcagatt ttccagccgt ctgctcttgc
    13501 tcagtctggc agacaggatg gccttggcgt ccacgccgct ggcgttgatg gggttttcct
    13561 cgaacagctg gttgtaggtc tgcaccagct ggatgaacag cttgtccacg tcgctgttgt
    13621 cggggttcag gtcgccctcg atcaggaagt ggccccggaa cttgatcatg tgggccaggg
    13681 ccagatagat cagccgcagg tcggccttgt cggtgctgtc caccagtttc tttctcaggt
    13741 ggtagatggt ggggtacttc tcgtggtagg ccacctcgtc cacgatgttg ccgaagatgg
    13801 ggtgccgctc gtgcttctta tcctcttcca ccaggaagga ctcttccagt ctgtggaaga
    13861 agctgtcgtc caccttggcc atctcgttgc tgaagatctc ttgcagatag cagatccggt
    13921 tcttccgtct ggtgtatctt cttctggcgg ttctcttcag ccgggtggcc tcggctgttt
    13981 cgccgctgtc gaacagcagg gctccgatca ggttcttctt gatgctgtgc cggtcggtgt
    14041 tgcccagcac cttgaatttc ttgctgggca ccttgtactc gtcggtgatc acggcccagc
    14101 ccacagagtt ggtgccgatg gccaggccga tgctgtactt cttgtccttt ttcttttttg
    14161 cctggccggc ctttttcgtg gccgccggcc ttttcatggt ggccgtacgt cacgacacct
    14221 gaaatggaag aaaaaaactt tgaaccactg tctgaggctt gagaatgaac caagatccaa
    14281 actcaaaaag ggcaaattcc aaggagaatt acatcaagtg ccaagctggc ctaacttcag
    14341 tctccaccca ctcagtgtgg ggaaactcca tcgcataaaa cccctccccc caacctaaag
    14401 acgacgtact ccaaaagctc gagaactaat cgaggtgcct ggacggcgcc cggtactccg
    14461 tggagtcaca tgaagcgacg gctgaggacg gaaaggccct tttcctttgt gtgggtgact
    14521 cacccgcccg ctctcccgag cgccgcgtcc tccattttga gctccctgca gcagggccgg
    14581 gaagcggcca tctttccgct cacgcaactg gtgccgaccg ggccagcctt gccgcccagg
    14641 gcggggcgat acacggcggc gcgaggccag gcaccagagc aggccggcca gcttgagact
    14701 acccccgtcc gattctcggt ggccgcgctc gcaggccccg cctcgccgaa catgtgcgct
    14761 gggacgcacg ggccccgtcg ccgcccgcgg ccccaaaaac cgaaatacca gtgtgcagat
    14821 cttggcccgc atttacaaga ctatcttgcc a
//
