LOCUS       Exported               11879 bp ds-DNA     circular SYN 19-JAN-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 11879)
  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 11879)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 19, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11879
                     /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
     primer_bind     1290..1352
                     /label=CS753_BsiWIsf_F61
     misc_feature    1309..1314
                     /label=BsiWI digest
     regulatory      1333..1342
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     1336..1355
                     /label=CS458_IRESNeon_F62.7
     CDS             1339..1977
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNG3K
                     /translation="MVSKGEEDNMASLPATHELHIFGSINDVDFDMVGQGTGNPNEGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVIGTGFPADGPVMTNTLTAADWCMSKMTYPNDKTIIS
                     TFKWSYITVNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSM"
     misc_feature    1978..2004
                     /label=twist fragment
     primer_bind     1980..1998
                     /label=CS393_MCS_F62.3
     misc_feature    1981..2033
                     /label=MCS
                     /note="multiple cloning site of fluorescent protein 
                     plasmids"
     primer_bind     complement(1996..2013)
                     /label=CS455_IRESmcs_R59.5
     misc_feature    2035..2621
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     2130..2149
                     /label=CS468_IRESchk_F61.6
     primer_bind     complement(2601..2624)
                     /label=CS450_ires_R61.2
     CDS             2610..2612
                     /codon_start=1
                     /product="start codon for translation from IRES2"
                     /label=ATG
                     /translation="M"
     CDS             2622..3317
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     primer_bind     3318..3340
                     /label=CS847_Scar2A_F64
     CDS             3327..3383
                     /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             3396..3989
                     /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     3886..3906
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    3996..4024
                     
     misc_feature    4014..4602
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4039..4063)
                     /label=CS408
     primer_bind     complement(4067..4087)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(4081..4100)
                     /label=WHV-5R
     CDS             complement(4485..4496)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4605..4621)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4606..4622)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5127..5307
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5333..5350)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5339..5563
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5609..6037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5696..5715)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5906..5927
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6046..6066)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6051..6380
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6231..6366
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6293..6312
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6428..6475
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6494..6868
                     /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     6701..6718
                     /label=CS555_endRuby_F61.3
     primer_bind     6701..6718
                     /label=CS555
     polyA_signal    6998..7119
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7035..7054)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7089..7108
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7168..7184)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7168..7184)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7181..7203)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7192..7208
                     /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(7216..7246)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7261..7282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7399..7416)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7570..8158)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7650..7669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8329..9189)
                     /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     8952..8971
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9190..9294)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9369..9388)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9560..9939
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9941..10139
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10094..10114
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             10157..10337
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10384..10509
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11002..11235
                     /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    11762..11879
                     /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 ggtgagcaag ggcgaggagg ataacatggc ctctctccca
     1381 gcgacacatg agttacacat ctttggctcc atcaacgatg tggactttga catggtgggt
     1441 cagggcaccg gcaatccaaa tgaaggttat gaggagttaa acctgaagtc caccaagggt
     1501 gacctccagt tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac
     1561 ctgccctacc ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggctac
     1621 caagtccatc gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac
     1681 acctacgagg gaagccacat caaaggagag gcccaggtga ttgggactgg tttccctgct
     1741 gacggtcctg tgatgaccaa caccctgacc gctgcggact ggtgcatgtc gaagatgact
     1801 taccccaacg acaaaaccat catcagtacc tttaagtgga gttacattac tgtgaatggc
     1861 aagcgctacc ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac
     1921 tatctgaaga accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccatgtaa
     1981 ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccgcccctc
     2041 tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     2101 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     2161 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     2221 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     2281 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     2341 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     2401 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     2461 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     2521 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     2581 ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc gagctgatta
     2641 aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac cacttcaagt
     2701 gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga atcaaggcgg
     2761 tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc atgtacggca
     2821 gcaaaacctt catcaaccac acccagggca tccccgactt ctttaagcag tccttccccg
     2881 agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg accgctaccc
     2941 aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga ggggtgaact
     3001 tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc tccaccgaga
     3061 ccctgtaccc cgctgacggc ggcctggaag gcagagccga catggccctg aagctcgtgg
     3121 gcgggggcca cctgatctgc aacttgaaga ccacatacag atccaagaaa cccgctaaga
     3181 acctcaagat gcccggcgtc tactatgtgg acagaagact ggaaagaatc aaggaggccg
     3241 acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc gacctcccta
     3301 gcaaactggg gcacagagga tctggtgcca cgaatttctc gctactcaag caggccggtg
     3361 atgtcgagga aaaccctggt cctatggtgt ctagagagta caagcccacg gtgcgcctcg
     3421 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     3481 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     3541 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     3601 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     3661 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     3721 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     3781 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     3841 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     3901 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     3961 ggtgcatgac ccgcaagccc ggtgcctgag aattcgatat caagcttatc ggtaatcaac
     4021 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     4081 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     4141 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     4201 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     4261 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     4321 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     4381 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     4441 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     4501 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4561 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     4621 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     4681 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     4741 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     4801 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
     4861 accacacaca aggctacttc cctgattggc agaactacac accagggcca gggatcagat
     4921 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag
     4981 aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg
     5041 acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg
     5101 cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg
     5161 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     5221 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     5281 tttagtcagt gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact
     5341 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5401 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     5461 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     5521 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     5581 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     5641 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5701 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5761 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5821 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5881 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5941 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     6001 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     6061 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6121 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     6181 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     6241 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6301 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     6361 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     6421 agcacgtgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     6481 gaggaactaa accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt
     6541 cgccggagcg gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga
     6601 cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca
     6661 ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc
     6721 cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga
     6781 gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt
     6841 gcacttcgtg gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc
     6901 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     6961 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     7021 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     7081 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     7141 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     7201 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     7261 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     7321 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7381 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7441 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7501 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7561 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7621 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7681 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7741 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7801 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7861 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7921 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7981 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     8041 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     8101 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     8161 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     8221 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     8281 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     8341 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8401 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8461 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8521 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8581 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8641 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8701 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8761 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8821 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8881 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8941 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     9001 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     9061 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     9121 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     9181 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     9241 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     9301 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     9361 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     9421 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     9481 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     9541 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     9601 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     9661 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     9721 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     9781 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     9841 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     9901 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     9961 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
    10021 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
    10081 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc
    10141 gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
    10201 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
    10261 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
    10321 gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag
    10381 gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg
    10441 cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc
    10501 gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg
    10561 ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa
    10621 cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga
    10681 cagctacaac catcccttca gacaggatca gaagaactta gatcattata taatacagta
    10741 gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac
    10801 aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt
    10861 cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt
    10921 agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag
    10981 agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag
    11041 cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat
    11101 agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact
    11161 cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa
    11221 ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt
    11281 gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg
    11341 gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga
    11401 atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag
    11461 tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat
    11521 agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt
    11581 taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga
    11641 caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt
    11701 agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca
    11761 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
    11821 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttt
//
