LOCUS       sequence-457033-        8756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mU6- and hU6-driven expression of dual RfxCas13d gRNAs and arrays 
            for combinatorial RNA knockdown..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8756)
  AUTHORS   El Kassem G, Hillmer J, Boettcher M
  TITLE     Evaluation of Cas13d as a tool for genetic interaction mapping.
  JOURNAL   Nat Commun. 2025 Feb 14;16(1):1631. doi: 10.1038/s41467-025-56747-4.
  PUBMED    39952934
REFERENCE   2  (bases 1 to 8756)
  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 
            Commun."; date: "2025-02-14"; volume: "16(1):1631. doi"; pages: " 
            10.1038/s41467-025-56747-4"
COMMENT     Sequence Label: 783-Rx-Dual: Combinatorial RfxCas13d gRNA and ...
FEATURES             Location/Qualifiers
     source          1..8756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(231..275)
                     /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    460..693
                     /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    1190..1315
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1362..1542
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(1560..1758)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1585..1605)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1760..2139
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2311..2330
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2405..2509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2510..2578,2579..3370)
                     /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(2728..2747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3541..4129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4030..4049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             4191..4371
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     4880..4896
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4881..4897
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4900..5488
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5006..5017
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5415..5435
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5568..5588)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(5598..6305)
                     /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(5873..5892)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6142..6160
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6302..6311
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(6318..6371)
                     /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(6381..6977)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6461..6481)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6958..6977
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6990..7201)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        complement(7304..7339)
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        complement(7461..7496)
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     promoter        complement(7500..7749)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(7558..7577)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7729..7749)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_RNA        complement(8006..8041)
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     misc_RNA        complement(8163..8198)
                     /label=RfxCas13d DR36
                     /note="36-base direct repeat for an RfxCas13d pre-gRNA 
                     (Konermann et al., 2018)"
     primer_bind     complement(8262..8282)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    8286..8319
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    8572..8689
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 atggatctgt ctctgtctct ctctccacct tcttcttcta ttccttcggg cctgtcgggt
       61 cccctcgggg ttgggaggtg ggtctgaaac gataatggtg aatatccctg cctaactcta
      121 ttcactatag aaagtacagc aaaaactatt cttaaaccta ccaagcctcc tactatcatt
      181 atgaataatt ttatatacca cagccaattt gttatgttaa accaattcca caaacttgcc
      241 catttatcta attccaataa ttcttgttca ttcttttctt gctggttttg cgattcttca
      301 attaaggagt gtattaagct tgtgtaattg ttaatttctc tgtcccactc catccaggtc
      361 gtgtgattcc aaatctgttc cagagattta ttactccaac tagcattcca aggcacagca
      421 gtggtgcaaa tgagttttcc agagcaaccc caaatcccca ggagctgttg atcctttagg
      481 tatctttcca cagccaggat tcttgcctgg agctgcttga tgccccagac tgtgagttgc
      541 aacagatgct gttgcgcctc aatagccctc agcaaattgt tctgctgctg cactatacca
      601 gacaataatt gtctggcctg taccgtcagc gtcattgacg ctgcgcccat agtgcttcct
      661 gctgctccca agaacccaag gaacaaagct cctattccca ctgctctttt ttctctctgc
      721 accactcttc tctttgcctt ggtgggtgct actcctaatg gttcaatttt tactacttta
      781 tatttatata attcacttct ccaattgtcc ctcatatctc ctcctccagg tctgaagatc
      841 agcggccggc cgcttgctgt gcggtggtct tacttttgtt ttgctcttcc tctatcttgt
      901 ctaaagcttc cttggtgtct tttatctcta tcctttgatg cacacaatag agggttgcta
      961 ctgtattata taatgatcta agttcttctg atcctgtctg aagggatggt tgtagctgtc
     1021 ccagtatttg tctacagcct tctgatgttt ctaacaggcc aggattaact gcgaatcgtt
     1081 ctagctccct gcttgcccat actatatgtt ttaatttata ttttttcttt ccccctggcc
     1141 ttaaccgaat tttttcccat cgcgatctaa ttctcccccg cttaatactg acgctctcgc
     1201 acccatctct ctccttctag cctccgctag tcaaaatttt tggcgtactc accagtcgcc
     1261 gcccctcgcc tcttgccgtg cgcgcttcag caagccgagt cctgcgtcga gagagctcct
     1321 ctggtttccc tttcgctttc aagtccctgt tcgggcgcca ctgctagaga ttttccacac
     1381 tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac
     1441 tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc
     1501 agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacagg caaaacgcgc
     1561 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     1621 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     1681 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     1741 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     1801 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     1861 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     1921 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     1981 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     2041 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     2101 taattgatta ctattaataa ctagtcaata atcaatgtca acgcgtatat ctggcccgta
     2161 catcgcgaag cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca
     2221 agccttgcct tgttgtagct taaattttgc tcgcgcacta ctcagcgacc tccaacacac
     2281 aagcagggag cagatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     2341 accatagggg atcgggagat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa
     2401 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2461 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     2521 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     2581 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     2641 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     2701 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     2761 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     2821 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     2881 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     2941 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     3001 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     3061 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     3121 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     3181 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     3241 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     3301 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     3361 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     3421 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3481 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     3541 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     3601 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     3661 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3721 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3781 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3841 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3901 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3961 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4021 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4081 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4141 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgc tgctagagat
     4201 tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg
     4261 ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc
     4321 tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc cagtacagtc
     4381 cggatgcagc tctcgggcca tgtgatgaaa tgctaggcgg ctgtcaaacc tccactctaa
     4441 tacttctctc tccgggtcat ccatcccatg caggctcaca gggtgtaaca agcgggtgtt
     4501 ctctccttca ttggcttctt ctaccttctc ttgctcaact ggtactagct tgtagcacca
     4561 tccaaaggtc agtggatatc tgatccctgg ccctggtgtg tagttctgcc aatcagggaa
     4621 gtagccttgt gtgtggtaga tccacagatc aaggatatct tgtcttcgtt gggagtgaat
     4681 tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag ctgccttgta
     4741 agtcattggt cttaaaggta cctgaggtgt gactggaaaa cccacctcct cctcctcttg
     4801 tgcttctagc caggcacaat cagcattggt agctgctgta ttgctacttg tgattgctcc
     4861 atgtttttct aggtctcgat cgaggtcgac ggtatcgatg cggggaggcg gcccaaaggg
     4921 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     4981 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     5041 cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg
     5101 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     5161 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     5221 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     5281 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     5341 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     5401 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     5461 cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat tgtacctagt
     5521 ggaaccggaa cccttaaaca tgttaggtcc ctcgacttaa ttaactatac ctttctcttc
     5581 ttttttggat cagctccctt gtacagctcg tccatgccgc cggtggagtg gcggccctcg
     5641 gcgcgttcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccaacttg
     5701 atgttgacgt tgtaggcgcc gggcagctgc acgggcttct tggccttgta ggtggtcttg
     5761 acctcagcgt cgtagtggcc gccgtccttc agcttcagcc tctgcttgat ctcgcccttc
     5821 agggcgccgt cctcggggta catccgctcg gaggaggcct cccagcccat ggtcttcttc
     5881 tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt gtagatgaac
     5941 tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc
     6001 atcacgcgct cccacttgaa gccctcgggg aaggacagct tcaagtagtc ggggatgtcg
     6061 gcggggtgct tcacgtaggc cttggagccg tacatgaact gaggggacag gatgtcccag
     6121 gcgaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag
     6181 gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg
     6241 tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc gcccttgctc
     6301 accatggtgg cgaccggtgg gccaggattc tcctcgacgt caccgcatgt tagcagactt
     6361 cctctgccct ctcccgatcc ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct
     6421 tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg
     6481 cggggcgcgg aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg
     6541 gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg
     6601 gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc
     6661 tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc
     6721 gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg
     6781 acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg
     6841 acccgctcga tgtggcggtc cgggtcgacg gtgtggcgcg tggcggggta gtcggcgaac
     6901 gcggcggcga gggtgcgtac ggcccggggg acgtcgtcgc gggtggcgag gcgcaccgtg
     6961 ggcttgtact cggtcatggt gggctagctc actgttctgg cggcaaaccc gttgcgaaaa
     7021 agaacgttca cggcgactac tgcacttata tacggttctc ccccaccctc gggaaaaagg
     7081 cggagccagt acacgacatc actttcccag tttaccccgc gccaccttct ctaggcaccg
     7141 gttcaattgc cgacccctcc ccccaacttc tcggggactg tgggcgatgt gcgctctgcc
     7201 cactgacggg cacatgcatg gcggtaatac ggttatccac gcgtgactgg agttcagacg
     7261 tgtgctcttc cgatctaaaa attgctagga ccggccttaa agcgtttcaa accccgacca
     7321 gttggtaggg gtttacttgg atcgcaggta catgtgccga gccatctctc ttaggcgctt
     7381 gctgctgcgc tgcggcgcgc ctcacaactc ctagctgtca ctcaagggaa gacgctggtg
     7441 ctggggaagt acctgcgtct gtttcaaacc ccgaccagtt ggtaggggtt tacttgcacg
     7501 gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt tcaagttacg
     7561 gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta cccaagaaat
     7621 tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt caaattaatt
     7681 ctaattatct ctctaacaga ccttgtatcg tatatgcaaa tatgaaggaa tcatgggaaa
     7741 taggccctct tcctgcccga ccgctaccta gggctcatca ccggcctctg gggcaaggtc
     7801 aatgtggccg aatgtggggc cgaagccctg gccaggctgc tgatcgtcta cccctggacc
     7861 cagaggttct ttgcgtcctt tgggaacctc tccagcccca ctgccatcct tggcaacccc
     7921 atggtccgcg cccacggcaa gaaagtgctc acctcctttg gggatgctgt gcatgcaaaa
     7981 aaaccttagc cgctaatagg tgagcgtttc aaaccccgac cagttggtag gggtttactt
     8041 ggatcgcagg tggttcctgg cttgcgtgct gggtggctcc ttcggctttg gcgcgcccct
     8101 tgggctctgc acctctgctg tgtcaggtgc cggcaccaac agcgccagtg acacctgcgt
     8161 ctgtttcaaa ccccgaccag ttggtagggg tttacttgcc aacaaggtgg ttctccaagg
     8221 gatacttata gtctcaaaac acacaattac tttacagtta gggtgagttt ccttttgtgc
     8281 tgtttataac ttcgtataat gtatgctata cgaagttata gaaatccaag cctatcatgt
     8341 aaaatgtagc tagtattaaa aagaacagat tatctgtctt ttatcgcaca ttaagcctct
     8401 atagttacta ggaaatatta tatgcaaatt aaccggggca ggggagtagc cgagcttctc
     8461 ccacaagtcc gtgcgagggg gccggcgcgg gcctagagat ggcgcgtcgg atctctagag
     8521 cgggcccggt actaaccaaa ctggatctct gctgtccctg taataaaccc gaaaattttg
     8581 aatttttgta atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac
     8641 tattctttcc cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa
     8701 tactgccatt tgtctgcaga attggcgcac gcagtgccga tccgttcact aatcga
//
