LOCUS       Exported               11036 bp ds-DNA     circular SYN 21-AUG-2024
DEFINITION  3rd generation lenti vector encoding dCas9 (S. pyogenes) fused with 
            VP64-p65-Rta (VPR) and 2A puromycin resistance marker; 
            EF1a-dCas9-VPR-P2A-Puro-WPRE).
ACCESSION   .
VERSION     .
KEYWORDS    lenti-EF1a-dCas9-VPR-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11036)
  AUTHORS   Ho SM, Hartley BJ, Flaherty E, Rajarajan P, Abdelaal R, Obiorah I, 
            Barretto N, Muhammad H, Phatnani HP, Akbarian S, Brennand KJ
  TITLE     Evaluating Synthetic Activation and Repression of 
            Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and 
            Astrocytes.
  JOURNAL   Stem Cell Reports. 2017 Aug 8;9(2):615-628. doi: 
            10.1016/j.stemcr.2017.06.012. Epub 2017 Jul 27.
  PUBMED    28757163
REFERENCE   2  (bases 1 to 11036)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Dec 11, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11036
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        130..1308
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          361..1299
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1256..1276
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1256..1276
                     /label=CS721
     misc_feature    1280..1309
                     
     misc_feature    1309..1314
                     /label=BsiWI digest
     misc_feature    1309
                     /label=first bp
     primer_bind     complement(1323..1365)
                     /label=CS999_chmpFLAG_R64
     regulatory      1333..1338
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     1339..1386
                     /label=CS1000_FLAGchmp_F63
     CDS             1342..1365
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1366..1527
                     /codon_start=1
                     /label=CHMP2B
                     /translation="MASLFKKKTVDDVIKEQNRELRGTQRAIIRDRAALEKQEKQLELE
                     IKKMAKIGN"
     CDS             1393..1521
                     /label=a1 by chmp2b similarity
     primer_bind     complement(1506..1554)
                     /label=CS685_chmpd5596_R59
     primer_bind     1528..1572
                     /label=CS686_chmpd5596_F59
     misc_feature    1528..1554
                     /label=GS
     primer_bind     1528..1545
                     /label=CS182
     primer_bind     complement(1537..1554)
                     /label=CS156
     CDS             1555..1905
                     /codon_start=1
                     /label=CHMP2B
                     /translation="KMAGAMSTTAKTMQAVNKKMDPQKTLQTMQNFQKENMKMEMTEEM
                     INDTLDDIFDGSDDEEESQDIVNQVLDEIGIEISGKMAKAPSAARSLPSASTSKATISD
                     EEIERQLKALGVD"
     CDS             1585..1608
                     /label=a3 by chmp2b similarity
     CDS             1624..1671
                     /label=a4 by chmp2a similarity
     CDS             1741..1788
                     /label=a5 by chmp2a similarity
     misc_feature    1759
                     /label=T instead of C for 165 DELTA
     CDS             1867..1899
                     /label=MIM Domain
     primer_bind     complement(1881..1933)
                     /label=CS764
     primer_bind     complement(1881..1905)
                     /label=CS945_CHMP2B2a_R64
     primer_bind     1908..1926
                     /label=CS393_MCS_F62.3
     misc_feature    1909..1961
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     misc_feature    1909..1932
                     /label=twist fragment
     primer_bind     complement(1924..1941)
                     /label=CS455_IRESmcs_R59.5
     primer_bind     1925..1942
                     /label=CS940_stab_F61
     misc_feature    1963..2549
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     2058..2077
                     /label=CS468_IRESchk_F61.6
     primer_bind     complement(2529..2552)
                     /label=CS450_ires_R61.2
     CDS             2538..2540
                     /codon_start=1
                     /product="start codon for translation from IRES2"
                     /label=ATG
                     /translation="M"
     regulatory      2550..2552
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2550..2552
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I (from SNCA-mScarlet)
                     /note="matures faster than mScarlet"
                     /translation="M"
     CDS             2553..3146
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="EYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVT
                     ELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQ
                     QQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAP
                     RNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2682..2699)
                     /label=CS964
     primer_bind     3043..3063
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3153..3181
                     
     misc_feature    3171..3759
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3196..3220)
                     /label=CS408
     primer_bind     complement(3224..3244)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(3238..3257)
                     /label=WHV-5R
     CDS             complement(3642..3653)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3762..3778)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3763..3779)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4284..4464
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4490..4507)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4496..4720
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4766..5194
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4853..4872)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5063..5084
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5203..5223)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5208..5537
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5388..5523
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5450..5469
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5585..5632
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5651..6025
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     5858..5875
                     /label=CS555_endRuby_F61.3
     primer_bind     5858..5875
                     /label=CS555
     polyA_signal    6155..6276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6192..6211)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6246..6265
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6325..6341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6325..6341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6338..6360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6349..6365
                     /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(6373..6403)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6418..6439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6556..6573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6727..7315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6807..6826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7486..8346)
                     /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     8109..8128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8347..8451)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8526..8545)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8717..9096
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9098..9296
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9251..9271
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             9314..9494
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9541..9666
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10159..10392
                     /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    10919..11036
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cgggtttatt acagggacag cagagatcca gtttggttaa ttagctagct gcaaagatgg
       61 ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga
      121 gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
      181 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
      241 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
      301 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
      361 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg
      421 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga
      481 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt
      541 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt
      601 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt
      661 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt
      721 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg
      781 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct
      841 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg
      901 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca
      961 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg
     1021 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg
     1081 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt
     1141 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac
     1201 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag
     1261 cctcagacag tggttcaaag tttttttctt ccatttcagg tgtcgtgacg tacgctagag
     1321 gatcgaaccc ttgccaccat ggactacaaa gacgacgatg ataaaatggc gtccctcttc
     1381 aagaagaaaa ccgtggatga tgtaataaag gaacagaatc gagagttacg aggtacacag
     1441 agggctataa tcagagatcg agcagcttta gagaaacaag aaaaacagct ggaattagaa
     1501 attaagaaaa tggccaagat tggtaatgga ggttcaggtt caggtggagg atccaagatg
     1561 gctggagcaa tgtctaccac agcaaaaaca atgcaggcag ttaacaagaa gatggatcca
     1621 caaaagacat tacaaacaat gcagaatttc cagaaggaaa acatgaaaat ggaaatgact
     1681 gaagaaatga tcaatgatac acttgatgac atctttgacg gttctgatga cgaagaagaa
     1741 agccaggata ttgtgaatca agttcttgat gaaattggaa ttgaaatttc tggaaagatg
     1801 gccaaagctc catcagctgc tcgaagctta ccatctgcct ctacttcaaa ggctacaatc
     1861 tcagatgaag agattgaacg gcaactcaag gctttaggag tagattaact cgagctcaag
     1921 cttcgaattc tgcagtcgac ggtaccgcgg gcccgggatc cgcccctctc cctccccccc
     1981 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     2041 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     2101 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     2161 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     2221 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     2281 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     2341 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     2401 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     2461 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     2521 aaaaacacga tgataatatg gccacaacca tggagtacaa gcccacggtg cgcctcgcca
     2581 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     2641 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     2701 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     2761 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     2821 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     2881 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     2941 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     3001 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     3061 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     3121 gcatgacccg caagcccggt gcctgagaat tcgatatcaa gcttatcggt aatcaacctc
     3181 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     3241 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     3301 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     3361 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     3421 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     3481 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     3541 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     3601 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     3661 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     3721 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag
     3781 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     3841 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     3901 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     3961 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     4021 acacacaagg ctacttccct gattggcaga actacacacc agggccaggg atcagatatc
     4081 cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag
     4141 ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc
     4201 cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc
     4261 gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc
     4321 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     4381 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     4441 agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg
     4501 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4561 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4621 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4681 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4741 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4801 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4861 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4921 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4981 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5041 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5101 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5161 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     5221 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5281 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5341 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5401 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5461 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5521 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     5581 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     5641 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
     5701 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
     5761 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
     5821 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
     5881 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
     5941 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
     6001 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     6061 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6121 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6181 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6241 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6301 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6361 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6421 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6481 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6541 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6601 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6661 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6721 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6781 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6841 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6901 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6961 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7021 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7081 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7141 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7201 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7261 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7321 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7381 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7441 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7501 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7561 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7621 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7681 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7741 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7801 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7861 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7921 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7981 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8041 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8101 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8161 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8221 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8281 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8341 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8401 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8461 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     8521 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     8581 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     8641 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     8701 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     8761 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     8821 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     8881 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     8941 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     9001 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     9061 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     9121 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     9181 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     9241 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg
     9301 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     9361 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     9421 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     9481 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
     9541 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     9601 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     9661 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     9721 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     9781 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     9841 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     9901 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     9961 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
    10021 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
    10081 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
    10141 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
    10201 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
    10261 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
    10321 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
    10381 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
    10441 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
    10501 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
    10561 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
    10621 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
    10681 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
    10741 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
    10801 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
    10861 gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt
    10921 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
    10981 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aatttt
//
