pCMV-T7-ABEmax(7.10)-SpCas9-NG-P2A-EGFP (RTW4564)

LOCUS       pCMV-T7-ABEmax(7.10)-SpCas9-NG-P2A-EGFP (RTW4564)               9627 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  CMV and T7 promoter expression plasmid for human codon optimized 
            ABEmax(7.10) A-to-G base editor with 
            SpCas9-NG(D10A/L1111R/D1135V/G1218R/E1219F/A1322R/R1335V/T1337R) and
            P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-T7-ABEmax(7.10)-SpCas9-NG-P2A-EGFP (RTW4564)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9627)
  AUTHORS   Walton RT, Christie KA, Whittaker MN, Kleinstiver BP
  TITLE     Unconstrained genome targeting with near-PAMless engineered 
            CRISPR-Cas9 variants.
  JOURNAL   Science. 2020 Apr 17;368(6488):290-296. doi: 
            10.1126/science.aba8853. Epub 2020 Mar 26.
  PUBMED    32217751
REFERENCE   2  (bases 1 to 9627)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..9627
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     521..540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(759..863)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(942..961)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1133..1512
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1513..1716
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1667..1687
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1758..1777
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1758..1776
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1826..1846
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3035..4750
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             7178..7198
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7208..7264
                     /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             7265..7981
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7310..7331)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7571..7590)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7918..7939
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7994..8014
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8023..8040
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8063..8080)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8069..8293
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8364..8380)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8364..8380)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8377..8399)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8388..8404
                     /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(8412..8442)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8457..8478
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8595..8612)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8766..9354)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8846..8865)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9525..758)
                     /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"
ORIGIN
        1 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
       61 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
      121 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
      181 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
      241 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
      301 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
      361 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
      421 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
      481 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
      541 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
      601 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
      661 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
      721 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
      781 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
      841 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
      901 tcgggagatc gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc
      961 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     1021 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     1081 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     1141 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     1201 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     1261 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     1321 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     1381 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     1441 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     1501 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     1561 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     1621 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     1681 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
     1741 tagagatccg cggccgctaa tacgactcac tatagggaga gccgccacca tgaaacggac
     1801 agccgacgga agcgagttcg agtcaccaaa gaagaagcgg aaagtctctg aagtcgagtt
     1861 tagccacgag tattggatga ggcacgcact gaccctggca aagcgagcat gggatgaaag
     1921 agaagtcccc gtgggcgccg tgctggtgca caacaataga gtgatcggag agggatggaa
     1981 caggccaatc ggccgccacg accctaccgc acacgcagag atcatggcac tgaggcaggg
     2041 aggcctggtc atgcagaatt accgcctgat cgatgccacc ctgtatgtga cactggagcc
     2101 atgcgtgatg tgcgcaggag caatgatcca cagcaggatc ggaagagtgg tgttcggagc
     2161 acgggacgcc aagaccggcg cagcaggctc cctgatggat gtgctgcacc accccggcat
     2221 gaaccaccgg gtggagatca cagagggaat cctggcagac gagtgcgccg ccctgctgag
     2281 cgatttcttt agaatgcgga gacaggagat caaggcccag aagaaggcac agagctccac
     2341 cgactctgga ggatctagcg gaggatcctc tggaagcgag acaccaggca caagcgagtc
     2401 cgccacacca gagagctccg gcggctcctc cggaggatcc tctgaggtgg agttttccca
     2461 cgagtactgg atgagacatg ccctgaccct ggccaagagg gcacgcgatg agagggaggt
     2521 gcctgtggga gccgtgctgg tgctgaacaa tagagtgatc ggcgagggct ggaacagagc
     2581 catcggcctg cacgacccaa cagcccatgc cgaaattatg gccctgagac agggcggcct
     2641 ggtcatgcag aactacagac tgattgacgc caccctgtac gtgacattcg agccttgcgt
     2701 gatgtgcgcc ggcgccatga tccactctag gatcggccgc gtggtgtttg gcgtgaggaa
     2761 cgcaaaaacc ggcgccgcag gctccctgat ggacgtgctg cactaccccg gcatgaatca
     2821 ccgcgtcgaa attaccgagg gaatcctggc agatgaatgt gccgccctgc tgtgctattt
     2881 ctttcggatg cctagacagg tgttcaatgc tcagaagaag gcccagagct ccaccgactc
     2941 cggaggatct agcggaggct cctctggctc tgagacacct ggcacaagcg agagcgcaac
     3001 acctgaaagc agcgggggca gcagcggggg gtcagacaag aagtacagca tcggcctggc
     3061 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     3121 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     3181 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     3241 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     3301 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     3361 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     3421 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     3481 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     3541 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     3601 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     3661 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     3721 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     3781 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     3841 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     3901 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     3961 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     4021 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     4081 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     4141 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     4201 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     4261 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     4321 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     4381 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     4441 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     4501 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     4561 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     4621 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     4681 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     4741 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     4801 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     4861 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     4921 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     4981 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     5041 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     5101 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     5161 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     5221 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     5281 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     5341 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     5401 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     5461 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     5521 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     5581 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     5641 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     5701 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     5761 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     5821 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     5881 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     5941 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     6001 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     6061 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     6121 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     6181 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     6241 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     6301 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     6361 cagacccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     6421 gtacggcggc ttcgtgagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     6481 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     6541 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     6601 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     6661 gaagagaatg ctggcctctg ccagattcct gcagaaggga aacgaactgg ccctgccctc
     6721 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     6781 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     6841 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     6901 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     6961 ccacctgttt accctgacca atctgggagc ccctagagcc ttcaagtact ttgacaccac
     7021 catcgaccgg aaggtgtaca gaagcaccaa agaggtgctg gacgccaccc tgatccacca
     7081 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg
     7141 cggctcaaaa agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcgg
     7201 aagcggagct actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg
     7261 acctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     7321 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     7381 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     7441 caccctcgtg accaccctga cctatggagt gcagtgcttc agccgctacc ccgaccacat
     7501 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     7561 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     7621 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     7681 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     7741 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     7801 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     7861 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     7921 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     7981 gtctggtggt tctcccaaga agaagaggaa agtctaaccg gtcatcatca ccatcaccat
     8041 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     8101 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     8161 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     8221 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     8281 gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct
     8341 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     8401 acaattccac acaacatacg agccggaagc ataaagtgta aagcctaggg tgcctaatga
     8461 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     8521 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     8581 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     8641 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     8701 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     8761 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     8821 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     8881 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     8941 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     9001 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     9061 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     9121 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     9181 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     9241 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     9301 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     9361 cctttgatct tttctacggg gtctgacact cagtggaacg aaaactcacg ttaagggatt
     9421 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     9481 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     9541 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     9601 gtcgtgtaga taactacgat acgggag
//