LOCUS       pCAG-eSpCas9-2A-GFP-sgRNA-RDH11       10246 bp    DNA     circular SYN 12-OCT-2020
DEFINITION  All-in-one CRISPR/Cas9 vector with high-fidelity eSpCas9 expression in human pluripotent stem cells.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-eCas9-GFP-U6-gRNA.
SOURCE      synthetic DNA construct.
  ORGANISM  synthetic DNA construct
FEATURES             Location/Qualifiers
     source          476..475
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /vntifkey="98"
     promoter        476..716
                     /vntifkey="30"
                     /label=U6\promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     476..496
                     /vntifkey="28"
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     647..666
                     /vntifkey="28"
                     /label=LKO.1\5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        746..821
                     /vntifkey="53"
                     /label=gRNA\scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system"
     enhancer        1001..1304
                     /vntifkey="9"
                     /label=CMV\enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1306..1583
                     /vntifkey="30"
                     /label=chicken\beta-actin\promoter
     intron          1585..2602
                     /vntifkey="15"
                     /label=chimeric\intron
                     /note="chimera between introns from chicken beta-actin and rabbit beta-globin"
     primer_bind     2526..2548
                     /vntifkey="28"
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2610..2629
                     /vntifkey="28"
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer"
     regulatory      2677..2686
                     /regulatory_class="other"
                     /vntifkey="21"
                     /label=Misc_Feature_1
                     /note="Unknown feature type:regulatory  vertebrate consensus sequence for strong initiation  of translation (Kozak, 1987)"
     CDS             2686..2751
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /vntifkey="4"
                     /label=3xFLAG
     CDS             2758..2778
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /vntifkey="4"
                     /label=SV40\NLS
     CDS             2803..6903
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting
                     specificity (Slaymaker et al., 2016)"
                     /vntifkey="4"
                     /label=eSpCas9(1.1)
                     /note="carries the mutations K848A, K1003A, and R1060A"
     CDS             6904..6951
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /vntifkey="4"
                     /label=nucleoplasmin\NLS
     CDS             6967..7020
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /vntifkey="4"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides."
     CDS             7021..7734
                     /codon_start=1
                     /product="enhanced GFP"
                     /vntifkey="4"
                     /label=EGFP
                     /note="mammalian codon-optimized"
     primer_bind     complement(7063..7084)
                     /vntifkey="28"
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7324..7343)
                     /vntifkey="28"
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7671..7692
                     /vntifkey="28"
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7762..7779)
                     /vntifkey="28"
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse"
     polyA_signal    7768..7975
                     /vntifkey="25"
                     /label=bGH\poly(A)\signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7984..8124
                     /vntifkey="34"
                     /label=AAV2\ITR
                     /note="inverted terminal repeat of adeno-associated virus serotype 2"
     repeat_region   7984..8113
                     /vntifkey="34"
                     /label=AAV2\ITR
     rep_origin      8199..8654
                     /direction=RIGHT
                     /vntifkey="33"
                     /label=f1\ori
                     /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
     primer_bind     complement(8286..8305)
                     /vntifkey="28"
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8496..8517
                     /vntifkey="28"
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8671..8690)
                     /vntifkey="28"
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable marker"
     primer_bind     8790..8812
                     /vntifkey="28"
                     /label=pGEX\3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8850..8868)
                     /vntifkey="28"
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward primer"
     promoter        8936..9040
                     /gene="bla"
                     /vntifkey="30"
                     /label=AmpR\promoter
     CDS             9041..9901
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /vntifkey="4"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and related antibiotics"
     primer_bind     complement(9259..9278)
                     /vntifkey="28"
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10072..414
                     /direction=RIGHT
                     /vntifkey="33"
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
     primer_bind     315..334
                     /vntifkey="28"
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    726..745
                     /vntifkey="21"
                     /label=RDH11\sgRNA
BASE COUNT     2507 a      2774 c      2938 g      2027 t 
ORIGIN
        1 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 
       61 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 
      121 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 
      181 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 
      241 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 
      301 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 
      361 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 
      421 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtgaggg 
      481 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat 
      541 tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta 
      601 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct 
      661 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa 
      721 caccgagttg atgtacacac cccacgtttt agagctagaa atagcaagtt aaaataaggc 
      781 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt ttagagctag 
      841 aaatagcaag ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa 
      901 atggctctag taacggccgc cagtgtgctg gaattcaggc tagttattaa tagtaatcaa 
      961 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa 
     1021 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 
     1081 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 
     1141 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 
     1201 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 
     1261 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca 
     1321 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta 
     1381 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg 
     1441 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc 
     1501 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata 
     1561 aaaagcgaag cgcgcggcgg gcggggagtc gctgcgacgc tgccttcgcc ccgtgccccg 
     1621 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt 
     1681 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct 
     1741 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg 
     1801 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc 
     1861 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag 
     1921 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg 
     1981 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc 
     2041 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc 
     2101 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg 
     2161 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg 
     2221 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct 
     2281 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc 
     2341 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc 
     2401 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct 
     2461 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag 
     2521 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca 
     2581 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca 
     2641 ttttggcaaa gaattcctga attctgcaga tccggtgcca ccatggacta taaggaccac 
     2701 gacggagact acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca 
     2761 aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc 
     2821 ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg 
     2881 cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg 
     2941 atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc 
     3001 gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc 
     3061 aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg 
     3121 gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc 
     3181 taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac 
     3241 aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac 
     3301 ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag 
     3361 ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac 
     3421 gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc 
     3481 cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc 
     3541 ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc 
     3601 aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc 
     3661 gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga 
     3721 gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag 
     3781 caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac 
     3841 aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc 
     3901 agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag 
     3961 gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac 
     4021 ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa 
     4081 gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc 
     4141 atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga 
     4201 aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc 
     4261 gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg 
     4321 ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg 
     4381 aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc 
     4441 atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac 
     4501 tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc 
     4561 aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg 
     4621 gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag 
     4681 gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg 
     4741 atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc 
     4801 aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc 
     4861 ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac 
     4921 atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg 
     4981 gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc 
     5041 gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac 
     5101 cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc 
     5161 atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag 
     5221 aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa 
     5281 ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg 
     5341 gccgacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc 
     5401 gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg 
     5461 aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc 
     5521 ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc 
     5581 acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac 
     5641 aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg 
     5701 aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc 
     5761 tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctgcgct ggaaagcgag 
     5821 ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag 
     5881 gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag 
     5941 accgagatta ccctggccaa cggcgagatc cggaaggcgc ctctgatcga gacaaacggc 
     6001 gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg 
     6061 agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa 
     6121 gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac 
     6181 cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc 
     6241 aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc 
     6301 atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac 
     6361 aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa 
     6421 aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc 
     6481 ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc 
     6541 tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac 
     6601 gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg 
     6661 gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag 
     6721 aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt 
     6781 gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg 
     6841 atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggc 
     6901 gacaaaaggc cggcggccac gaaaaaggcc ggccaggcaa aaaagaaaaa ggaattcggc 
     6961 agtggagagg gcagaggaag tctgctaaca tgcggtgacg tcgaggagaa tcctggccca 
     7021 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 
     7081 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 
     7141 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 
     7201 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag 
     7261 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 
     7321 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 
     7381 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 
     7441 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 
     7501 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 
     7561 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 
     7621 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg 
     7681 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caaggaattc 
     7741 taactagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 
     7801 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 
     7861 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 
     7921 tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct ggggagcggc 
     7981 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg 
     8041 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg 
     8101 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt 
     8161 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt 
     8221 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc 
     8281 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 
     8341 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 
     8401 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 
     8461 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 
     8521 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt 
     8581 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt 
     8641 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag 
     8701 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc 
     8761 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc 
     8821 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa 
     8881 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg 
     8941 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 
     9001 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 
     9061 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 
     9121 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 
     9181 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 
     9241 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga 
     9301 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 
     9361 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 
     9421 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 
     9481 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 
     9541 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac 
     9601 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 
     9661 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 
     9721 gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca ttgcagcact 
     9781 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 
     9841 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 
     9901 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 
     9961 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 
    10021 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 
    10081 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 
    10141 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 
    10201 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccac 
//
