LOCUS       sequence-324507-       12995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of S. pyogenes sgRNA with a GFP-P2A-PuroR 
            selection marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12995)
  AUTHORS   Black JB, McCutcheon SR, Dube S, Barrera A, Klann TS, Rice GA, Adkar
            SS, Soderling SH, Reddy TE, Gersbach CA
  TITLE     Master Regulators and Cofactors of Human Neuronal Cell Fate 
            Specification Identified by CRISPR Gene Activation Screens.
  JOURNAL   Cell Rep. 2020 Dec 1;33(9):108460. doi: 
            10.1016/j.celrep.2020.108460.
  PUBMED    33264623
REFERENCE   2  (bases 1 to 12995)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2020-12-1"; volume: "33(9):108460. doi"; pages: " 
            10.1016/j.celrep.2020.108460"
COMMENT     Sequence Label: pLV_hU6-sgRNA_hUbC-GFP-P2A-PuroR
FEATURES             Location/Qualifiers
     source          1..12995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        225..423
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     378..398
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             441..621
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    668..793
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1286..1519
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1704..1748
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2046..2163
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2208..2224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2243..2263
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2245..2263
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        complement(4264..4512)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4322..4341)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4492..4512)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4526..4545)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4527..4545)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4555..4572)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4555..4571)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4600..5811
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5648..5667
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             join(5840..5842,5843..5845,5846..6556)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5840..5842,5843..5845,5846..6556)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5885..5906)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6146..6165)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6493..6514
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6566..6622
                     /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             6623..7222
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6623..6642)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7119..7139
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(7235..7268)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    7293..7881
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7346..7366)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7764..7775)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7884..7900)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7885..7901)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8406..8586
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(8612..8629)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8618..8842
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8888..9316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8975..8994)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9185..9206
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9325..9345)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9330..9659
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9510..9645
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9572..9591
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9707..9754
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9773..10147
                     /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"
     polyA_signal    10277..10398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10314..10333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10368..10387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10447..10463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10460..10482)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10471..10487
                     /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(join(10495..10501,10502..10519,10520..10525))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10540..10561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10678..10695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10849..11437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10929..10948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11608..12399,12400..12468))
                     /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     12231..12250
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12469..12573)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12648..12667)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(12839..12995,1..223)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
       61 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      121 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      181 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      241 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      301 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      361 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      421 cagcgcgttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
      481 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      541 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      601 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
      661 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
      721 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
      781 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
      841 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
      901 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
      961 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     1021 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     1081 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     1141 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     1201 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     1261 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     1321 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     1381 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     1441 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     1501 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     1561 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     1621 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     1681 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     1741 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     1801 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     1861 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     1921 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     1981 gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc
     2041 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     2101 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     2161 tttcgggttt attacaggga cagcagagat ccagtttggt taatacacag gaaacagcta
     2221 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     2281 ccaccgcgga aaaaaagcac cgactcggtg ccactttttc aagttgataa cggactagcc
     2341 ttatttaaac ttgctatgct gtttccagca tagctcttaa acagagacgt acaaaaaaga
     2401 gcaagaagct aaaaaagatt taaaaattat ttttagcgca gttaatggaa caggaactaa
     2461 atttacccca aaaatattac gtgaatcagg atataacgtt attgaggttg aagagcatgc
     2521 atttgaagat gaaacattta aaaatgttgt aaatccaaat ccagaatttg atcctgcatg
     2581 aaaaataccg cttgaatatg gtattaaaca tgatgcagat attattatta tgaatgaccc
     2641 agatgctgac agatttggaa tggcaataaa acatgatggt cattttgtaa gattagatgg
     2701 aaatcaaaca ggaccaattt taattgattg aaaattatca aatctaaaac gcttaaatag
     2761 cattccaaaa aatccggctc tatattcaag ttttgtaaca agtgatttgg gtgatagaat
     2821 cgctcatgaa aaatatggag ttaatattgt aaaaacttta actggattta aatgaatggg
     2881 tagagaaatt gctaaagaag aagataacgg attaaatttt gtttttgctt atgaagaaag
     2941 ttatggatat gtaattgatg actcagctag agataaagat ggaatacaag cttctatatt
     3001 aatagcagag gctgcttgat tttataaaaa acaaaataaa acattagtag actatttaga
     3061 agatttattt aaagaaatgg gtgcatatta cactttcact ttaaacttga attttaaacc
     3121 agaagaaaag aaattaaaaa ttgaaccatt aatgaaatca ttgagagcaa cacccttaac
     3181 tcaaattgct ggacttaaag ttgttaatgt tgaagactac atcgatggaa tgtataatat
     3241 gccaggacaa gacttactaa aattttattt agaagataag tcatgatttg ctgttcgccc
     3301 aagtggaact gaacctaaac taaaaattta ttttataggt gttggtgaat ctgttcaaaa
     3361 cgctaaagtt aaagtagacg aaattattaa agaattaaaa ttaaaaatga atatatagga
     3421 gaaaaaatga aactaaacaa atatatagat cacacattat taaaacaaga tgctacgaaa
     3481 gctgaaatta aacaattatg tgatgaagca attgaatttg attttgcaac agtttgtgtt
     3541 aattcatatt gaacaagcta ttgtaaagaa ttattaaaag gcacaaatgt aggaataaca
     3601 aatgttgtag gttttcctct aggtgcatgc acaacagcta caaaagcatt cgaagtttct
     3661 gaagcaatta aagatggtgc aacagaaatt gatatggtat taaatattgg tgcattaaaa
     3721 gacaaaaatt atgaattagt tttagaagac atgaaagctg taaaaaaagc agctggatca
     3781 catgttgtta aatgtattat ggaaaattgt ttattaacaa aagaagaaat catgaaagct
     3841 tgtgaaatag ctgttgaagc tggattagaa tttgttaaaa catcaacagg attttcaaaa
     3901 tcaggtgcaa catttgaaga tgttaaacta atgaagtcag ttgttaaaga caatgcttta
     3961 gttaaagcag ctggtggagt tagaacattt gaagatgctc aaaaaatgat tgaagcagga
     4021 gctgaccgct taggaacaag tggtggagta gctattatta aaggtgaaga aaacaacgcg
     4081 agttactaaa actagcgttt ttttattttg ctcattttta ttaaaagttt gcaaaaagga
     4141 acataaaaat tctaattatt gatactaaag ttattaaaaa gaagattttg gttgatttta
     4201 taaaggtcat agaatataat attttagcat gtgtattttg tgtgctcatt tacaaccgtc
     4261 tccggtgttt cgtcctttcc acaagatata taaagccaag aaatcgaaat actttcaagt
     4321 tacggtaagc atatgatagt ccattttaaa acataatttt aaaactgcaa actacccaag
     4381 aaattattac tttctacgtc acgtattttg tactaatatc tttgtgttta cagtcaaatt
     4441 aattccaatt atctctctaa cagccttgta tcgtatatgc aaatatgaag gaatcatggg
     4501 aaataggccc tcggtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     4561 cgtcgtttta caacattaac ccgtgtcggc tccagatctg gcctccgcgc cgggttttgg
     4621 cgcctcccgc gggcgccccc ctcctcacgg cgagcgctgc cacgtcagac gaagggcgca
     4681 gcgagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact
     4741 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta
     4801 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg
     4861 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg
     4921 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
     4981 gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc
     5041 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
     5101 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc
     5161 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
     5221 ggcatggtgg gcggcaagaa cccaaggtct tgaggccttc gctaatgcgg gaaagctctt
     5281 attcgggtga gatgggctgg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     5341 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     5401 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     5461 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     5521 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     5581 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     5641 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     5701 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     5761 actagtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga cgaagcttgg
     5821 gctgcaggtc gactctagaa tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     5881 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     5941 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     6001 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     6061 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     6121 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     6181 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     6241 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     6301 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     6361 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     6421 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     6481 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     6541 ggacgagctg tacaagggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga
     6601 tgtcgaagag aatcctggac cgatgaccga gtacaagccc acggtgcgcc tcgccacccg
     6661 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     6721 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     6781 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     6841 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     6901 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     6961 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tgtcgcccga
     7021 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     7081 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     7141 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     7201 gacccgcaag cccggtgcct aaagttgata atagataact tcgtatagca tacattatac
     7261 gaagttatga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     7321 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     7381 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     7441 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     7501 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     7561 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     7621 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     7681 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     7741 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     7801 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     7861 ccctttgggc cgcctccccg catcgatacc gtcgacctcg agacctagaa aaacatggag
     7921 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
     7981 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
     8041 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     8101 actcccaacg aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc
     8161 ctgattggca gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt
     8221 gctacaagct agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca
     8281 cccgcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag
     8341 agtggaggtt tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact
     8401 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     8461 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     8521 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     8581 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     8641 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     8701 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     8761 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     8821 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     8881 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     8941 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     9001 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     9061 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     9121 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     9181 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     9241 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     9301 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     9361 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     9421 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     9481 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     9541 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     9601 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     9661 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     9721 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     9781 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     9841 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     9901 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     9961 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
    10021 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
    10081 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
    10141 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
    10201 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
    10261 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
    10321 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
    10381 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
    10441 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
    10501 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
    10561 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
    10621 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
    10681 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
    10741 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
    10801 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
    10861 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
    10921 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
    10981 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
    11041 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
    11101 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
    11161 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
    11221 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
    11281 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
    11341 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
    11401 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    11461 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
    11521 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
    11581 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
    11641 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
    11701 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
    11761 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
    11821 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
    11881 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
    11941 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    12001 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    12061 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    12121 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    12181 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    12241 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    12301 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    12361 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    12421 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    12481 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    12541 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    12601 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
    12661 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
    12721 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
    12781 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
    12841 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
    12901 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
    12961 gacccccgcc cattgacgtc aataatgacg tatgt
//
