LOCUS       sequence-425605-       11739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lenti vector encoding the MS2-P65-HSF1 activator helper complex with
            a 2A Hygro resistance marker (EF1a-MS2-p65-HSF1-2A-Hygro-WPRE). This
            version has a higher virus titer..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11739)
  AUTHORS   Joung J, Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham
            MD, Sanjana NE, Zhang F
  TITLE     Genome-scale CRISPR-Cas9 knockout and transcriptional activation 
            screening.
  JOURNAL   Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. 
            Epub 2017 Mar 23.
  PUBMED    28333914
REFERENCE   2  (bases 1 to 11739)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nprot.2017.016"; journalName: "Nat Protoc"; date: 
            "2017-04"; volume: "12"; issue: "4"; pages: "828-863"
COMMENT     Sequence Label: lentiMPH v2
FEATURES             Location/Qualifiers
     source          1..11739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    481..606
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             653..833
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(851..1049)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(876..896)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1051..1430
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1602..1621
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1696..1800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1801..1869,1870..2661)
                     /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(2019..2038)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2832..3420
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3321..3340
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3574..3591
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3708..3729
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3744..3749,3750..3767,3768..3774)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3782..3798
                     /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     3787..3809
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3806..3822
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3806..3822
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3871..3992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3882..3901)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3936..3955
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4122..4496)
                     /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(4515..4562)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(4610..4939)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4624..4759
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4678..4697)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4924..4944
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4953..5381)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5063..5084)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5275..5294
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5427..5651
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5640..5657
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             5683..5863
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     6368..6384
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     6369..6385
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6468..7646
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6699..7637
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7594..7614
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7653..7662
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7662..8048
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             8103..8123
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8139..8681
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             8706..9077
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             9093..9146
                     /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             9150..10169
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     misc_feature    10209..10797
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10262..10282)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10680..10691)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    10850..10967
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(11265..11309)
                     /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    11494..11727
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 cttttttctc tctgcaccac tcttctcttt gccttggtgg gtgctactcc taatggttca
       61 atttttacta ctttatattt atataattca cttctccaat tgtccctcat atctcctcct
      121 ccaggtctga agatcagcgg ccgcttgctg tgcggtggtc ttacttttgt tttgctcttc
      181 ctctatcttg tctaaagctt ccttggtgtc ttttatctct atcctttgat gcacacaata
      241 gagggttgct actgtattat ataatgatct aagttcttct gatcctgtct gaagggatgg
      301 ttgtagctgt cccagtattt gtctacagcc ttctgatgtt tctaacaggc caggattaac
      361 tgcgaatcgt tctagctccc tgcttgccca tactatatgt tttaatttat attttttctt
      421 tccccctggc cttaaccgaa ttttttccca tcgcgatcta attctccccc gcttaatact
      481 gacgctctcg cacccatctc tctccttcta gcctccgcta gtcaaaattt ttggcgtact
      541 caccagtcgc cgcccctcgc ctcttgccgt gcgcgcttca gcaagccgag tcctgcgtcg
      601 agagagctcc tctggtttcc ctttcgcttt caagtccctg ttcgggcgcc actgctagag
      661 attttccaca ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga
      721 cgggcacaca ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc
      781 cctagttagc cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag
      841 gcaaaacgcg ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa
      901 tggggcggag ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact
      961 cccattgacg tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg
     1021 cccattgatg tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat
     1081 gtactgccaa gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca
     1141 tttaccgtca ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc
     1201 caagtgggca gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg
     1261 cgttactatg ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca
     1321 gccaggcggg ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac
     1381 taatgacccc gtaattgatt actattaata actagtcaat aatcaatgtc aacgcgtata
     1441 tctggcccgt acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca
     1501 attgtcggtc aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac
     1561 ctccaacaca caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt
     1621 actgagagtg caccataggg gatcgggaga tctcccgatc cgtcgacgtc aggtggcact
     1681 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     1741 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     1801 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     1861 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1921 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1981 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2041 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2101 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2161 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2221 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2281 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2341 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2401 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2461 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2521 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2581 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2641 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     2701 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     2761 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     2821 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     2881 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2941 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3001 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3061 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3121 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3181 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3241 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3301 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3361 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3421 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3481 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3541 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     3601 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     3661 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     3721 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     3781 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     3841 tagctagagg tcgacggtat acagacatga taagatacat tgatgagttt ggacaaacca
     3901 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3961 ttgtaaccat tataagctgc aataaacaag ttggggtggg cgaagaactc cagcatgaga
     4021 tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa gcccaacctt
     4081 tcatagaagg cggcggtgga atcgaaatct cgtagcacgt gtcagtcctg ctcctcggcc
     4141 acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc ccacggctgc
     4201 tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga cacgacctcc
     4261 gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag ggtgttgtcc
     4321 ggcaccacct ggtcctggac cgcgctgatg aacagggtca cgtcgtcccg gaccacaccg
     4381 gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc ggtcggtcca gaactcgacc
     4441 gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac tggtcaactt ggccatggtt
     4501 tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga ttaattgtca
     4561 acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc
     4621 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct
     4681 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     4741 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     4801 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     4861 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     4921 cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt gttaaatcag
     4981 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5041 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5101 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5161 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5221 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5281 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5341 caccacaccc gccgcgctta atgcgccgct acagggcgcg tggggatacc ccctagagcc
     5401 ccagctggtt ctttccgcct cagaagccat agagcccacc gcatccccag catgcctgct
     5461 attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg
     5521 acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt
     5581 ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact
     5641 agaaggcaca gtcgaggctg atcagcgggt ttaaacgggc cctgctagag attttccaca
     5701 ctgactaaaa gggtctgagg gatctctagt taccagagtc acacaacaga cgggcacaca
     5761 ctacttgaag cactcaaggc aagctttatt gaggcttaag cagtgggttc cctagttagc
     5821 cagagagctc ccaggctcag atctggtcta accagagaga cccagtacag tccggatgca
     5881 gctctcgggc catgtgatga aatgctaggc ggctgtcaaa cctccactct aatacttctc
     5941 tctccgggtc atccatccca tgcaggctca cagggtgtaa caagcgggtg ttctctcctt
     6001 cattggcttc ttctaccttc tcttgctcaa ctggtactag cttgtagcac catccaaagg
     6061 tcagtggata tctgatccct ggccctggtg tgtagttctg ccaatcaggg aagtagcctt
     6121 gtgtgtggta gatccacaga tcaaggatat cttgtcttcg ttgggagtga attagccctt
     6181 ccagtccccc cttttctttt aaaaagtggc taagatctac agctgccttg taagtcattg
     6241 gtcttaaagg tacctgaggt gtgactggaa aacccacctc ctcctcctct tgtgcttcta
     6301 gccaggcaca atcagcattg gtagctgctg tattgctact tgtgattgct ccatgttttt
     6361 ctaggtctcg aggtcgacgg tatcgattgc aaagatggat aaagttttaa acagagagga
     6421 atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc tccggtgccc
     6481 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     6541 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     6601 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     6661 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc
     6721 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc
     6781 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc
     6841 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg
     6901 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc
     6961 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg
     7021 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg
     7081 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga
     7141 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc
     7201 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg
     7261 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atgaaggacg cggcgctcgg
     7321 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg
     7381 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct
     7441 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca
     7501 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat
     7561 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt
     7621 tttttcttcc atttcaggtg tcgtgacgta cggccaccat ggcttcaaac tttactcagt
     7681 tcgtgctcgt ggacaatggt gggacagggg atgtgacagt ggctccttct aatttcgcta
     7741 atggggtggc agagtggatc agctccaact cacggagcca ggcctacaag gtgacatgca
     7801 gcgtcaggca gtctagtgcc cagaagagaa agtataccat caaggtggag gtccccaaag
     7861 tggctaccca gacagtgggc ggagtcgaac tgcctgtcgc cgcttggagg tcctacctga
     7921 acatggagct cactatccca attttcgcta ccaattctga ctgtgaactc atcgtgaagg
     7981 caatgcaggg gctcctcaaa gacggtaatc ctatcccttc cgccatcgcc gctaactcag
     8041 gtatctacag cgctggagga ggtggaagcg gaggaggagg aagcggagga ggaggtagcg
     8101 gacctaagaa aaagaggaag gtggcggccg ctggatcccc ttcagggcag atcagcaacc
     8161 aggccctggc tctggcccct agctccgctc cagtgctggc ccagactatg gtgccctcta
     8221 gtgctatggt gcctctggcc cagccacctg ctccagcccc tgtgctgacc ccaggaccac
     8281 cccagtcact gagcgctcca gtgcccaagt ctacacaggc cggcgagggg actctgagtg
     8341 aagctctgct gcacctgcag ttcgacgctg atgaggacct gggagctctg ctggggaaca
     8401 gcaccgatcc cggagtgttc acagatctgg cctccgtgga caactctgag tttcagcagc
     8461 tgctgaatca gggcgtgtcc atgtctcata gtacagccga accaatgctg atggagtacc
     8521 ccgaagccat tacccggctg gtgaccggca gccagcggcc ccccgacccc gctccaactc
     8581 ccctgggaac cagcggcctg cctaatgggc tgtccggaga tgaagacttc tcaagcatcg
     8641 ctgatatgga ctttagtgcc ctgctgtcac agatttcctc tagtgggcag ggaggaggtg
     8701 gaagcggctt cagcgtggac accagtgccc tgctggacct gttcagcccc tcggtgaccg
     8761 tgcccgacat gagcctgcct gaccttgaca gcagcctggc cagtatccaa gagctcctgt
     8821 ctccccagga gccccccagg cctcccgagg cagagaacag cagcccggat tcagggaagc
     8881 agctggtgca ctacacagcg cagccgctgt tcctgctgga ccccggctcc gtggacaccg
     8941 ggagcaacga cctgccggtg ctgtttgagc tgggagaggg ctcctacttc tccgaagggg
     9001 acggcttcgc cgaggacccc accatctccc tgctgacagg ctcggagcct cccaaagcca
     9061 aggaccccac tgtctcctgt acaggcagtg gagagggcag aggaagtctg ctaacatgcg
     9121 gtgacgtcga ggagaatcct ggcccaacca tgaaaaagcc tgaactcacc gctacctctg
     9181 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctccgagg
     9241 gcgaagaatc tcgggctttc agcttcgatg tgggagggcg tggatatgtc ctgcgggtga
     9301 atagctgcgc cgatggtttc tacaaagatc gctatgttta tcggcacttt gcatccgccg
     9361 ctctccctat tcccgaagtg cttgacattg gggagttcag cgagagcctg acctattgca
     9421 tctcccgccg tgcacagggt gtcaccttgc aagacctgcc tgaaaccgaa ctgcccgctg
     9481 ttctccagcc cgtcgccgag gccatggatg ccatcgctgc cgccgatctt agccagacca
     9541 gcgggttcgg cccattcgga cctcaaggaa tcggtcaata cactacatgg cgcgatttca
     9601 tctgcgctat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     9661 gtgcctccgt cgcccaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     9721 tccggcacct cgtgcacgcc gatttcggct ccaacaatgt cctgaccgac aatggccgca
     9781 taacagccgt cattgactgg agcgaggcca tgttcgggga ttcccaatac gaggtcgcca
     9841 acatcttctt ctggaggccc tggttggctt gtatggagca gcagacccgc tacttcgagc
     9901 ggaggcatcc cgagcttgca ggatctcctc ggctccgggc ttatatgctc cgcattggtc
     9961 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggctcagg
    10021 gtcgctgcga cgcaatcgtc cggtccggag ccgggactgt cgggcgtaca caaatcgccc
    10081 gcagaagcgc tgccgtctgg accgatggct gtgtggaagt gctcgccgat agtggaaaca
    10141 gacgccccag cactcgtcct agggcaaagg atctgcagta atgagaattc gatatcaagc
    10201 ttatcggtaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
    10261 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
    10321 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
    10381 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
    10441 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
    10501 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
    10561 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
    10621 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
    10681 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
    10741 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
    10801 gctagctaat taaccaaact ggatctctgc tgtccctgta ataaacccga aaattttgaa
    10861 tttttgtaat ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta
    10921 ttctttcccc tgcactgtac cccccaatcc ccccttttct tttaaaattg tggatgaata
    10981 ctgccatttg tctgcagaat tggcgcacgc agtgccgatc cgttcactaa tcgaatggat
    11041 ctgtctctgt ctctctctcc accttcttct tctattcctt cgggcctgtc gggtcccctc
    11101 ggggttggga ggtgggtctg aaacgataat ggtgaatatc cctgcctaac tctattcact
    11161 atagaaagta cagcaaaaac tattcttaaa cctaccaagc ctcctactat cattatgaat
    11221 aattttatat accacagcca atttgttatg ttaaaccaat tccacaaact tgcccattta
    11281 tctaattcca ataattcttg ttcattcttt tcttgctggt tttgcgattc ttcaattaag
    11341 gagtgtatta agcttgtgta attgttaatt tctctgtccc actccatcca ggtcgtgtga
    11401 ttccaaatct gttccagaga tttattactc caactagcat tccaaggcac agcagtggtg
    11461 caaatgagtt ttccagagca accccaaatc cccaggagct gttgatcctt taggtatctt
    11521 tccacagcca ggattcttgc ctggagctgc ttgatgcccc agactgtgag ttgcaacaga
    11581 tgctgttgcg cctcaatagc cctcagcaaa ttgttctgct gctgcactat accagacaat
    11641 aattgtctgg cctgtaccgt cagcgtcatt gacgctgcgc ccatagtgct tcctgctgct
    11701 cccaagaacc caaggaacaa agctcctatt cccactgct
//
