LOCUS       sequence-392596-        9841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPRa reporter cloning backbone and MCP-p65-HSF1-2A-Puro 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9841)
  AUTHORS   Ishiguro S, Ishida K, Sakata RC, Ichiraku M, Takimoto R, Yogo R, 
            Kijima Y, Mori H, Tanaka M, King S, Tarumoto S, Tsujimura T, Bashth 
            O, Masuyama N, Adel A, Toyoshima H, Seki M, Oh JH, Archambault AS, 
            Nishida K, Kondo A, Kuhara S, Aburatani H, Klein Geltink RI, 
            Yamamoto T, Shakiba N, Takashima Y, Yachie N
  TITLE     A multi-kingdom genetic barcoding system for precise clone 
            isolation.
  JOURNAL   Nat Biotechnol. 2025 May 21. doi: 10.1038/s41587-025-02649-1.
  PUBMED    40399693
REFERENCE   2  (bases 1 to 9841)
  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"; journalName: "Nat 
            Biotechnol. 2025 May 21. doi: 10.1038/s41587-025-02649-1."
COMMENT     Sequence Label: pLV-CS-060
FEATURES             Location/Qualifiers
     source          1..9841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(189..293)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    341..475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(378..397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     432..451
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             643..1276
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1323..1448
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1945..2178
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2363..2407
                     /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    2670..2785
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     CDS             complement(2833..3543)
                     /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(3111..3130)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3380..3398
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3540..3549
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3579..3617)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        3696..3734
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3764..3773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3770..3772,3773..3775,3776..4489)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3770..3772,3773..3775,3776..4486)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3815..3836)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4076..4095)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4423..4444
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        4542..4753
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             4764..5150
                     /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             5205..5225
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5241..5783
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             5808..6179
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             6186..6242
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6243..6839
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6736..6756
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6852..7440
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6905..6925)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7323..7334)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7647..7880
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8008..8024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8008..8024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8021..8043)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8032..8048
                     /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(8056..8062,8063..8080,8081..8086))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8101..8122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8239..8256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8410..8998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8490..8509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9169..9841,1..119,120..188))
                     /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     9792..9811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
       61 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      121 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      181 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      301 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc aacttgttta ttgcagctta
      361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
      421 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcaactg
      481 gataactcaa gctaaccaaa atcatcccaa acttcccacc ccatacccta ttaccactgc
      541 caattacctg tggtttcatt tactctaaac ctgtgattcc tctgaattat tttcatttta
      601 aagaaattgt atttgttaaa tatgtactac aaacttagta gttggaaggg ctaattcact
      661 cccaaagaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
      721 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
      781 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
      841 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
      901 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
      961 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
     1021 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     1081 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     1141 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     1201 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     1261 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     1321 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     1381 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     1441 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     1501 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     1561 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     1621 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     1681 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     1741 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc ggccgctgat
     1801 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
     1861 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
     1921 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
     1981 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
     2041 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
     2101 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
     2161 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
     2221 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
     2281 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
     2341 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
     2401 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
     2461 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
     2521 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
     2581 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
     2641 attagtgaac ggatctcgac ggtatcgcct ttaaaagaaa aggggggatt ggggggtaca
     2701 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     2761 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
     2821 atcgatttaa atttacttgt acagctcgtc catgccgccg gtggagtggc ggccctcggc
     2881 gcgttcgtac tgttccacga tggtgtagtc ctcgttgtgg gaggtgatgt ccaacttgat
     2941 gttgacgttg taggcgccgg gcagctgcac gggcttcttg gccttgtagg tggtcttgac
     3001 ctcagcgtcg tagtggccgc cgtccttcag cttcagcctc tgcttgatct cgcccttcag
     3061 ggcgccgtcc tcggggtaca tccgctcgga ggaggcctcc cagcccatgg tcttcttctg
     3121 cattacgggg ccgtcggagg ggaagttggt gccgcgcagc ttcaccttgt agatgaactc
     3181 gccgtcctgc agggaggagt cctgggtcac ggtcaccacg ccgccgtcct cgaagttcat
     3241 cacgcgctcc cacttgaagc cctcggggaa ggacagcttc aagtagtcgg ggatgtcggc
     3301 ggggtgcttc acgtaggcct tggagccgta catgaactga ggggacagga tgtcccaggc
     3361 gaagggcagg gggccaccct tggtcacctt cagcttggcg gtctgggtgc cctcgtaggg
     3421 gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg ttcacggagc cctccatgtg
     3481 caccttgaag cgcatgaact ccttgatgat ggccatgtta tcctcctcgc ccttgctcac
     3541 catggtggcg ttaacgcgat ctgacggttc actaaacgag ctctgcttat ataggcctcc
     3601 caccgtacac gcctacctac tgactgcagt ctgagtctga cagcctctag agtgttgtaa
     3661 agctagcgga gcagagctac gcactctacg ctagaggtag gcgtgtacgg tgggaggcct
     3721 atataagcag agctcgttta gtgaaccgtc agatcgcttc gaagccacca tggtgagcaa
     3781 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3841 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3901 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3961 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     4021 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     4081 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4141 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4201 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     4261 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     4321 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     4381 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4441 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat taattaacta
     4501 ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     4561 cacagtcccc gagaagttgg ggggaggggt cggcaattga tccggtgcct agagaaggtg
     4621 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     4681 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     4741 cgccagaaca cagggaattc atggcttcaa actttactca gttcgtgctc gtggacaatg
     4801 gtgggacagg ggatgtgaca gtggctcctt ctaatttcgc taatggggtg gcagagtgga
     4861 tcagctccaa ctcacggagc caggcctaca aggtgacatg cagcgtcagg cagtctagtg
     4921 cccagaagag aaagtatacc atcaaggtgg aggtccccaa agtggctacc cagacagtgg
     4981 gcggagtcga actgcctgtc gccgcttgga ggtcctacct gaacatggag ctcactatcc
     5041 caattttcgc taccaattct gactgtgaac tcatcgtgaa ggcaatgcag gggctcctca
     5101 aagacggtaa tcctatccct tccgccatcg ccgctaactc aggtatctac agcgctggag
     5161 gaggtggaag cggaggagga ggaagcggag gaggaggtag cggacctaag aaaaagagga
     5221 aggtggcggc cgctggatcc ccttcagggc agatcagcaa ccaggccctg gctctggccc
     5281 ctagctccgc tccagtgctg gcccagacta tggtgccctc tagtgctatg gtgcctctgg
     5341 cccagccacc tgctccagcc cctgtgctga ccccaggacc accccagtca ctgagcgctc
     5401 cagtgcccaa gtctacacag gccggcgagg ggactctgag tgaagctctg ctgcacctgc
     5461 agttcgacgc tgatgaggac ctgggagctc tgctggggaa cagcaccgat cccggagtgt
     5521 tcacagatct ggcctccgtg gacaactctg agtttcagca gctgctgaat cagggcgtgt
     5581 ccatgtctca tagtacagcc gaaccaatgc tgatggagta ccccgaagcc attacccggc
     5641 tggtgaccgg cagccagcgg ccccccgacc ccgctccaac tcccctggga accagcggcc
     5701 tgcctaatgg gctgtccgga gatgaagact tctcaagcat cgctgatatg gactttagtg
     5761 ccctgctgtc acagatttcc tctagtgggc agggaggagg tggaagcggc ttcagcgtgg
     5821 acaccagtgc cctgctggac ctgttcagcc cctcggtgac cgtgcccgac atgagcctgc
     5881 ctgaccttga cagcagcctg gccagtatcc aagagctcct gtctccccag gagcccccca
     5941 ggcctcccga ggcagagaac agcagcccgg attcagggaa gcagctggtg cactacacag
     6001 cgcagccgct gttcctgctg gaccccggct ccgtggacac cgggagcaac gacctgccgg
     6061 tgctgtttga gctgggagag ggctcctact tctccgaagg ggacggcttc gccgaggacc
     6121 ccaccatctc cctgctgaca ggctcggagc ctcccaaagc caaggacccc actgtctcct
     6181 gtacagcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac
     6241 cgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     6301 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     6361 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     6421 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     6481 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     6541 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     6601 ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag ggtctgggca
     6661 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     6721 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     6781 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac
     6841 gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     6901 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6961 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     7021 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     7081 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     7141 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     7201 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     7261 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     7321 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     7381 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     7441 ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     7501 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     7561 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     7621 ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga
     7681 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     7741 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     7801 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     7861 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     7921 tgcaaagaaa tgaatatcag agagtgagag gccttgacat tctagcgttt taccgtcgac
     7981 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     8041 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     8101 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     8161 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     8221 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     8281 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     8341 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     8401 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     8461 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     8521 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     8581 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     8641 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     8701 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     8761 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     8821 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     8881 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8941 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     9001 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     9061 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     9121 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     9181 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     9241 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     9301 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     9361 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     9421 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     9481 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     9541 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     9601 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     9661 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     9721 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     9781 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9841 a
//
