pAsCpf1(TATV)(BB) (pY221)

LOCUS       pAsCpf1(TATV)(BB) (pY221)                8435 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Expresses humanized AsCpf1 TATV PAM variant and crRNA guide.
ACCESSION   .
VERSION     .
KEYWORDS    pAsCpf1(TATV)(BB) (pY221)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8435)
  AUTHORS   Gao L, Cox DBT, Yan WX, Manteiga JC, Schneider MW, Yamano T, 
            Nishimasu H, Nureki O, Crosetto N, Zhang F
  TITLE     Engineered Cpf1 variants with altered PAM specificities.
  JOURNAL   Nat Biotechnol. 2017 Jun 5. doi: 10.1038/nbt.3900.
  PUBMED    28581492
REFERENCE   2  (bases 1 to 8435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8435
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        114..399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        complement(531..771)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(581..600)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(751..771)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   784..913
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(975..1563)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1055..1074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1734..2594)
                     /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     2357..2376
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2595..2699)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2767..2785
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2823..2845)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2945..2964
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2981..3436)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3118..3139)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3330..3349
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   3511..3651
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3522..3651
                     /label=AAV2 ITR
     polyA_signal    3660..3867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3856..3873
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(3901..3981)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             complement(3988..4035)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(4036..7953)
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNVEKNRGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      4307..4316
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(7978..7998)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      8001..8010
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     intron          8042..8269
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     promoter        complement(8270..112)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ccccccgccc ccatcgctgc acaaaataat taaaaaataa ataaatacaa aattgggggt
       61 ggggaggggg gggagatggg gagagtgaag cagaacgtgg ggctcacctc gaccatggta
      121 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
      181 tgggcacaat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
      241 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
      301 cgtcaatgga aagtccctat tggcgttact attgacgtca atgggcgggg gtcgttgggc
      361 ggtcagccag gcgggccatt taccgtaagt tatgtaacgg gtacccgcgt atatctggcc
      421 cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat gcaatagaaa
      481 aaaaggtctt ctcgaagacc catctacaag agtagaaatt acggtgtttc gtcctttcca
      541 caagatatat aaagccaaga aatcgaaata ctttcaagtt acggtaagca tatgatagtc
      601 cattttaaaa cataatttta aaactgcaaa ctacccaaga aattattact ttctacgtca
      661 cgtattttgt actaatatct ttgtgtttac agtcaaatta attccaatta tctctctaac
      721 agccttgtat cgtatatgca aatatgaagg aatcatggga aataggccct ctctagaggc
      781 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
      841 aggccgggcg accaaaggtc gcccgacgcc cgggcggcct cagtgagcga gcgagcgcgc
      901 agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
      961 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1021 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1081 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1141 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1201 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1261 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1321 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1381 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     1441 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     1501 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     1561 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     1621 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     1681 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     1741 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     1801 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     1861 gcaatgatac cgcggcttcc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     1921 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     1981 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     2041 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     2101 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     2161 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     2221 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     2281 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     2341 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     2401 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     2461 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     2521 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     2581 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     2641 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     2701 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     2761 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     2821 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2881 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     2941 tatgcggcat cagagcagat tgtactgaga gtgcaccata aaattgtaaa cgttaatatt
     3001 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca atagaccgaa
     3061 atcggcaaaa tcccttataa atcaaaagaa tagcccgaga tagagttgag tgttgttcca
     3121 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3181 gtctatcagg gcgatggccc actacgtgaa ccatcaccca aatcaagttt tttggggtcg
     3241 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3301 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctaag
     3361 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3421 ccgctacagg gcgcgtacta tggttgcttt gacgtatgcg gtgtgaaata ccgcacagat
     3481 gcgtaaggag aaaataccgc atcaggcgcc cctgcaggca gctgcgcgct cgctcgctca
     3541 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     3601 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgct
     3661 ccccagcatg cctgctattc tcttcccaat cctccccctt gctgtcctgc cccaccccac
     3721 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
     3781 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
     3841 cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgagctcta ggaattctta
     3901 ggcatagtcg gggacatcat atgggtatgc ataatcaggc acgtcgtagg gataagcgta
     3961 atctggaaca tcgtatgggt aggatccctt tttctttttt gcctggccgg cctttttcgt
     4021 ggccgccggc cttttgttgc gcagctcctg gatgtaggcc agccagtcct gattggagat
     4081 gccgttctgc agcttcagat ccttgctctc cttcaggtga ttcagcagca gctggccctt
     4141 cagggcgatg tggtaggcgc cattggcatc ggcgtccatg ggccactctg ggttctgaaa
     4201 ccgggagtcg aagcacacgc cattcagatc gcgcacgggg ctgttgatat agtcctcgcc
     4261 tgtggcggca ttggagttcc gcatctgcag cacgctgcgg atcagggcca ccatggtgtc
     4321 gatggcgtga gaatcgtcat tctccagcag ctttggcagg atgttggagc catccctgaa
     4381 cacgatgccc ttctcctcca gcagggcgat cagctcgttg gcaggataca ggtcccggta
     4441 tctgccggtg aatctgtgat tctcgatcac tggcacgatt ctcttgccgg cgatgaaagg
     4501 ggtgcccttg gcgtcaaact gtgtctcgtt cttctcgaac acgatatccc atgcaggcat
     4561 aaagccgggc aggcccctct ggaaggacag atttctgttc atcttaaagt gcaggatgaa
     4621 gtcgccggtt ttcacgtcgt agtgcagaaa gtcgaagccc tccaggaagt gcttgcggct
     4681 ctcgtgattc ttgatggttt tccacacgaa ggggtccacg aagccggtca ggggatcgat
     4741 cttagatgta tatggggcag gcacgtaaaa caggaagcca gactgggtgc ccatcttggc
     4801 aaaggaggtg aactggtctg tcagctggta tgggttcagc acgcctccca ctttctctgc
     4861 tggatagtcc ttcagcacca ggcaattcag cttatcgatc agcatcttct cgaactgctg
     4921 gtacacggcc ttctcggcga tgccggtcct cttgctctta aagccgaaat tcaggttctc
     4981 cagcaccacc acggcctggt agtggatcat caggtccacg atctcgtgga tgacctggct
     5041 cagatagccc tgcttcagat ccttgattgt gcccaccaca gaccaggcct gccttgctgc
     5101 caccctctcc ttctccctgt tgtccagctt cttctggtaa tcaaactgct ggatggtgtt
     5161 caggctccgc tgctccagga tcttgccggt ggagtcgatc actgtgatat agatcaggtt
     5221 tctctcgccc cgatcgatgc cgatgatagg tgtctcgggg tgctccttca ggtaggcatt
     5281 caccctctgg ttgaacttag atggggaatt ggcggcctga tagttcagtg tgataggcac
     5341 gtggaaaaag aacttgtcgc tggtaaagcg cctatccttg atgatctcgt gagacacctc
     5401 cttggtgatc acgttgggca gcagggccct ggcctcatca gacaggtcgt gggacagtct
     5461 gtgattcaca tagtcgtaca gctcctggta cagggtgtcg gggattgggg ttttctgatc
     5521 cttcagcttc ttgttcagca tcttctctcc cagccggtgt gccatcctct tcatcctgga
     5581 cttagggcgg tagaacagct cggcctggcc attcagcttg atgcttgtct tggccaggtt
     5641 ctctggagaa aacagaccgg tccaatacag tgtgtgcaga ttaggcttgc cgtggtggcc
     5701 cttggcaaag tccttgttat agatctggaa caggtacagc ttgcctgtct ccacggcatc
     5761 catgatctcc ttctcggcga ttctctggaa gctgatgtgg tacagcaggg gattcagctc
     5821 ggcatagtac tcgcccaggt ccttatactg agaggatggc cgcaggctag acagatcgat
     5881 agaggttgtc ttggtatact tggacagaaa atcccttgtg aagtcgatcc acttgcacag
     5941 ggcctctctg tagcccttct ggtcgccggt tttcttggcg tacgctgtct gaaacttctt
     6001 tggctccttc tcaggattgt tcaggtcgta gatctccttt gtgatctcca gaggctcgat
     6061 gaaattgttg gacagcagga tgggggttgt gtgggtctga aagtgggcgg tcaccgcctt
     6121 cagctgggtg ctgcactttg ggatcatctt ggcggcatcc ggaaagtagt catagtacat
     6181 cttatcaaag ccctcgctgg ttttctctgt gggctcgaag ctgagggcct tatacctgcc
     6241 cttctgcttt ggcatgatgc ccagatagta caggccgttc ttcacaaaca ggatggcgcc
     6301 ccggttcttc tccacattca cgtcccagcc ccgggccagt gtaggcatct gaaagttcag
     6361 cttgaacttc tccacggagt agggcttctt ggtggcataa tttctggcct tgttgtagaa
     6421 gctcagagaa ggctccatct ccagcttgat gccggtcagc cgggcagaga actcggggtc
     6481 cacctcgttg gactcatcca cggcaaacca gtccagcagg tggtacaggc ccagcaggct
     6541 gtccagctga gacttcagga tctccttctc ctcctgcttc ttcagggttg taggcagtgg
     6601 ctgatccagg gcggcgtgtg cgtgggacag gatctcgctg gttttctgct tgaaggcctc
     6661 gctcagctcc ttgcctgcgg cagagatgat ctcctgcagg ttgatatcct cgtgcttcag
     6721 gctgcgctgc accttctcct tggcagactt ggtgatcttg cctgtcagct cggagattct
     6781 ccgctcatac agggcattcc tcagtgtatc ccagtggtcg cacagggcgc tgctgattgt
     6841 ctccagcttc ttgtggctga tgaagatgtg tgtcaggtcg atgctgttca gctcgttaaa
     6901 cagggcctcg gctgtctcca gcacgttctc gtttctcagc agtgtcttgt acttgcagaa
     6961 ggactggatc acttcctcgt cgctcttaaa ctcctccagg atgaaagaca gggtgttcct
     7021 atcggacagg atctgcttaa acagggggat gaatctgtgt ggcagggagg cgatgatgtg
     7081 ggctgtctca tcattcttct ggatggccag attcagcacc tcgttcaggc ccttgatctt
     7141 ctcggtgcct gcctcccgag agattcctcc cagcagctgg ttatacaggt cgatctgggt
     7201 ctgtgtcagc agctggttat aaaaagggaa ggaaaacacc tcctcgatgg aggtgctcac
     7261 gaagatgccg atggccttct tcacgttctc aaagtgctcc cgcaggctgg gcacggcggt
     7321 gatcaggcgt gtgaagatgt gacaattctc cttaaacttg gggaagttgt cctgcacgat
     7381 gcggtgtggg atggctgtgc tgatatcctc ggcgctgaac acgttcttcc tgttctcata
     7441 aaagccggag aagtaggttg taaacttgtc gaagctccgc agcagggcgt tctcgtgctc
     7501 ggttgtggtc acggtgccca gctgcttcag caccttgcca ttaaacagct cggccttgaa
     7561 caggcccttg tagatctcgg cgtgtctctt attgatggca tcggtcaggt tgtctgtccg
     7621 gccgatgaag tagtcgtgga tggcattgcg atatgtggcc tgctcctcga tcagggcgtt
     7681 ccttgtctcc tcggttttct cctttctata ggagtcgatg gcggcgctca ggttctccca
     7741 atccagctgc accagctgca ggcactggtc ggcataggtc ttgtagatcc gatcgatgat
     7801 gggcttcagc tccttgtagt gatcattgcg ggccttgtcc tcctcgatga agccctgctc
     7861 ctggatgtgc ttcagggtct tgccctgtgg gatcagctca aaccgcagtg tcttgctcac
     7921 ctgatacagg ttggtaaagc cctcgaactg tgtggctgct gggactccgt ggataccgac
     7981 cttccgcttc ttctttgggg ccatggtggc aagcttaagt ttaaacgcta gccggtccaa
     8041 cctgaaaaaa agtgatttca ggcaggtgct ccaggtaatt aaacattaat accccaccaa
     8101 ccaaccatcc cttaaaccct tacctcttgc tcagctaatt acagcccgga ggagaagggc
     8161 cgtcccgccc gctcacctgt gggagtaacg cggtcagtca gagccggggc gggcggcgcg
     8221 aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc agcgactccc gcccgccgcg
     8281 cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga
     8341 gcgcgccgct ctgattggct gccgccgcac ctctccgcct cgccccgccc cgcccctcgc
     8401 cccgccccgc cccgcctggc gcgcgccccc ccccc
//