LOCUS       sequence-450821-        8221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CBh-SaCas9-2A-GFP, and hU6-sgRNA (Sa) with a BbsI golden gate 
            cloning backbone for sgRNA. sgRNA uses 3TC scaffold for increased 
            sgRNA expression. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8221)
  AUTHORS   Chey YCJ, Gierus L, Lushington C, Arudkumar JC, B Geiger A, Staker 
            LG, Robertson LJ, Pfitzner C, Kennedy JG, Lee RHB, Godahewa GI, 
            Adikusuma F, Thomas PQ
  TITLE     Optimal SpCas9- and SaCas9-mediated gene editing by enhancing gRNA 
            transcript levels through scaffold poly-T tract reduction.
  JOURNAL   BMC Genomics. 2025 Feb 12;26(1):138. doi: 
            10.1186/s12864-025-11317-2.
  PUBMED    39939860
REFERENCE   2  (bases 1 to 8221)
  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: "BMC 
            Genomics."; date: "2025-02-12"; volume: "26(1):138. doi"; pages: " 
            10.1186/s12864-025-11317-2"
COMMENT     Sequence Label: SaCas9 GFP V3
FEATURES             Location/Qualifiers
     source          1..8221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1979..2026
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             2042..2095
                     /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             join(2096..2098,2099..2809)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2096..2098,2099..2809)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2138..2159)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2399..2418)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2746..2767
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2837..2854)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2843..3050
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3059..3199
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3059..3188
                     /label=AAV2 ITR
     rep_origin      3274..3729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3361..3380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3571..3592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3746..3765)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3865..3887
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3925..3943)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4011..4115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4116..4184,4185..4976)
                     /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(4334..4353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5147..5735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5636..5655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        5797..6045
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5797..5817
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     5968..5987
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6064..6139
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     enhancer        6160..6445
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          6748..6975
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      6987..6996
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6999..7019
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(7044..8221,1..1978)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
ORIGIN
        1 cctgagcctg aaggccatca acctgatcct ggacgagctg tggcacacca acgacaacca
       61 gatcgctatc ttcaaccggc tgaagctggt gcccaagaag gtggacctgt cccagcagaa
      121 agagatcccc accaccctgg tggacgactt catcctgagc cccgtcgtga agagaagctt
      181 catccagagc atcaaagtga tcaacgccat catcaagaag tacggcctgc ccaacgacat
      241 cattatcgag ctggcccgcg agaagaactc caaggacgcc cagaaaatga tcaacgagat
      301 gcagaagcgg aaccggcaga ccaacgagcg gatcgaggaa atcatccgga ccaccggcaa
      361 agagaacgcc aagtacctga tcgagaagat caagctgcac gacatgcagg aaggcaagtg
      421 cctgtacagc ctggaagcca tccctctgga agatctgctg aacaacccct tcaactatga
      481 ggtggaccac atcatcccca gaagcgtgtc cttcgacaac agcttcaaca acaaggtgct
      541 cgtgaagcag gaagaaaaca gcaagaaggg caaccggacc ccattccagt acctgagcag
      601 cagcgacagc aagatcagct acgaaacctt caagaagcac atcctgaatc tggccaaggg
      661 caagggcaga atcagcaaga ccaagaaaga gtatctgctg gaagaacggg acatcaacag
      721 gttctccgtg cagaaagact tcatcaaccg gaacctggtg gataccagat acgccaccag
      781 aggcctgatg aacctgctgc ggagctactt cagagtgaac aacctggacg tgaaagtgaa
      841 gtccatcaat ggcggcttca ccagctttct gcggcggaag tggaagttta agaaagagcg
      901 gaacaagggg tacaagcacc acgccgagga cgccctgatc attgccaacg ccgatttcat
      961 cttcaaagag tggaagaaac tggacaaggc caaaaaagtg atggaaaacc agatgttcga
     1021 ggaaaagcag gccgagagca tgcccgagat cgaaaccgag caggagtaca aagagatctt
     1081 catcaccccc caccagatca agcacattaa ggacttcaag gactacaagt acagccaccg
     1141 ggtggacaag aagcctaata gagagctgat taacgacacc ctgtactcca cccggaagga
     1201 cgacaagggc aacaccctga tcgtgaacaa tctgaacggc ctgtacgaca aggacaatga
     1261 caagctgaaa aagctgatca acaagagccc cgaaaagctg ctgatgtacc accacgaccc
     1321 ccagacctac cagaaactga agctgattat ggaacagtac ggcgacgaga agaatcccct
     1381 gtacaagtac tacgaggaaa ccgggaacta cctgaccaag tactccaaaa aggacaacgg
     1441 ccccgtgatc aagaagatta agtattacgg caacaaactg aacgcccatc tggacatcac
     1501 cgacgactac cccaacagca gaaacaaggt cgtgaagctg tccctgaagc cctacagatt
     1561 cgacgtgtac ctggacaatg gcgtgtacaa gttcgtgacc gtgaagaatc tggatgtgat
     1621 caaaaaagaa aactactacg aagtgaatag caagtgctat gaggaagcta agaagctgaa
     1681 gaagatcagc aaccaggccg agtttatcgc ctccttctac aacaacgatc tgatcaagat
     1741 caacggcgag ctgtatagag tgatcggcgt gaacaacgac ctgctgaacc ggatcgaagt
     1801 gaacatgatc gacatcacct accgcgagta cctggaaaac atgaacgaca agaggccccc
     1861 caggatcatt aagacaatcg cctccaagac ccagagcatt aagaagtaca gcacagacat
     1921 tctgggcaac ctgtatgaag tgaaatctaa gaagcaccct cagatcatca aaaagggcaa
     1981 aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat tcggcagtgg
     2041 agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag
     2101 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2161 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2221 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2281 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2341 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2401 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2461 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2521 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     2581 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     2641 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     2701 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2761 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg aattctaact
     2821 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2881 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2941 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3001 caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag
     3061 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     3121 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     3181 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3241 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     3301 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     3361 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3421 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3481 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3541 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3601 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     3661 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     3721 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     3781 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     3841 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     3901 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3961 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4021 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4081 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4141 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4201 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4261 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4321 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4381 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4441 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4501 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4561 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4621 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4681 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4741 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4801 ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc
     4861 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4921 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4981 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5041 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5101 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5161 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5221 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5281 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5341 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5401 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5461 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5521 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5581 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5641 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5701 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5761 ggttcctggc cttttgctgg ccttttgctc acatgtgagg gcctatttcc catgattcct
     5821 tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta
     5881 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     5941 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     6001 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgggtc ttcgagaaga
     6061 cctgtttcag tactctggaa acagaatcta ctgaaacaag gcaaaatgcc gtgtttatct
     6121 cgtcaacttg ttggcgagat tttttgcggc cgcggtaccc gttacataac ttacggtaaa
     6181 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag taacgccaat
     6241 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     6301 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     6361 cgcctggcat tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6421 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
     6481 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     6541 agcgatgggg gcgggggggg gggggggggg gcgccggggg gggggggggg gggggggggg
     6601 gggggggggg gggggggcgg gggggggggg ggggcgaggc ggagaggtgc ggcggcagcc
     6661 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
     6721 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
     6781 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     6841 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg
     6901 tttaagggat ggttggttgg tggggtatta atgtttaatt acctggagca cctgcctgaa
     6961 atcacttttt ttcaggttgg accggtgcca ccatggcccc aaagaagaag cggaaggtcg
     7021 gtatccacgg agtcccagca gccaagcgga actacatcct gggcctggac atcggcatca
     7081 ccagcgtggg ctacggcatc atcgactacg agacacggga cgtgatcgat gccggcgtgc
     7141 ggctgttcaa agaggccaac gtggaaaaca acgagggcag gcggagcaag agaggcgcca
     7201 gaaggctgaa gcggcggagg cggcatagaa tccagagagt gaagaagctg ctgttcgact
     7261 acaacctgct gaccgaccac agcgagctga gcggcatcaa cccctacgag gccagagtga
     7321 agggcctgag ccagaagctg agcgaggaag agttctctgc cgccctgctg cacctggcca
     7381 agagaagagg cgtgcacaac gtgaacgagg tggaagagga caccggcaac gagctgtcca
     7441 ccaaagagca gatcagccgg aacagcaagg ccctggaaga gaaatacgtg gccgaactgc
     7501 agctggaacg gctgaagaaa gacggcgaag tgcggggcag catcaacaga ttcaagacca
     7561 gcgactacgt gaaagaagcc aaacagctgc tgaaggtgca gaaggcctac caccagctgg
     7621 accagagctt catcgacacc tacatcgacc tgctggaaac ccggcggacc tactatgagg
     7681 gacctggcga gggcagcccc ttcggctgga aggacatcaa agaatggtac gagatgctga
     7741 tgggccactg cacctacttc cccgaggaac tgcggagcgt gaagtacgcc tacaacgccg
     7801 acctgtacaa cgccctgaac gacctgaaca atctcgtgat caccagggac gagaacgaga
     7861 agctggaata ttacgagaag ttccagatca tcgagaacgt gttcaagcag aagaagaagc
     7921 ccaccctgaa gcagatcgcc aaagaaatcc tcgtgaacga agaggatatt aagggctaca
     7981 gagtgaccag caccggcaag cccgagttca ccaacctgaa ggtgtaccac gacatcaagg
     8041 acattaccgc ccggaaagag attattgaga acgccgagct gctggatcag attgccaaga
     8101 tcctgaccat ctaccagagc agcgaggaca tccaggaaga actgaccaat ctgaactccg
     8161 agctgaccca ggaagagatc gagcagatct ctaatctgaa gggctatacc ggcacccaca
     8221 a
//
