LOCUS       sequence-462960-       12986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral plasmid for cloning an spCas9 gRNA with puromycin 
            resistance and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12986)
  AUTHORS   Rohm D, Black JB, McCutcheon SR, Barrera A, Berry SS, Morone DJ, 
            Nuttle X, de Esch CE, Tai DJC, Talkowski ME, Iglesias N, Gersbach CA
  TITLE     Activation of the imprinted Prader-Willi syndrome locus by 
            CRISPR-based epigenome editing.
  JOURNAL   Cell Genom. 2025 Feb 12;5(2):100770. doi: 
            10.1016/j.xgen.2025.100770.
  PUBMED    39947136
REFERENCE   2  (bases 1 to 12986)
  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 
            Genom."; date: "2025-02-12"; volume: "5(2):100770. doi"; pages: " 
            10.1016/j.xgen.2025.100770"
COMMENT     Sequence Label: pLV-U6-gRNA scaffold-UbC-mCherry-PuroR
FEATURES             Location/Qualifiers
     source          1..12986
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     329..348
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        423..527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(528..596,597..1388)
                     /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     complement(746..765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1559..2147
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2048..2067
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2301..2318
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2435..2456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2471..2476,2477..2494,2495..2501)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2509..2525
                     /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)."
     primer_bind     2514..2536
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2533..2549
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2598..2719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2609..2628)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2663..2682
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2849..3223)
                     /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"
     promoter        complement(3242..3289)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(3337..3666)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3351..3486
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3405..3424)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     3651..3671
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3680..4108)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3790..3811)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4002..4021
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    4154..4378
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4367..4384
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             4410..4590
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     5095..5111
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5096..5112
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5115..5703
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5221..5232
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5630..5650
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    5728..5761
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(5774..6373)
                     /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(5857..5877)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6354..6373
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(6374..6430)
                     /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             complement(6440..7147)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6715..6734)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6984..7002
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(7176..8387)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(7320..7339)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     primer_bind     8415..8432
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8416..8432
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8442..8461
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8442..8460
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        8475..8723
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8475..8495
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8646..8665
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(10724..10744)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10724..10742)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10763..10779)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    10824..10941
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(11239..11283)
                     /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    11468..11701
                     /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    12194..12319
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             12366..12546
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(12564..12762)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(12589..12609)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(12764..12986,1..157)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
       61 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
      121 attgattact attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca
      181 tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag
      241 ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa
      301 gcagggagca gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
      361 cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg
      421 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      481 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      541 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      601 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      661 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      721 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
      781 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
      841 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
      901 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
      961 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1021 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1081 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1141 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1201 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1261 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1321 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1381 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1441 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1501 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1561 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1621 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1681 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1741 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1801 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1861 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1921 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1981 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2041 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2101 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2161 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2221 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2281 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2341 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     2401 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     2461 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     2521 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     2581 acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2641 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2701 aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga
     2761 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     2821 cggtggaatc gaaatctcgt agcacgtgtc agtcctgctc ctcggccacg aagtgcacgc
     2881 agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg ccgatctcgg
     2941 tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac cactcggcgt
     3001 acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc accacctggt
     3061 cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg aagtcgtcct
     3121 ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt
     3181 cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catggtttag ttcctcacct
     3241 tgtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca cgtgctgatc
     3301 agatccgaaa atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa
     3361 aaagcctcct cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata
     3421 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     3481 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     3541 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
     3601 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
     3661 acacacattc cacagaatta attcgcgtta aatttttgtt aaatcagctc attttttaac
     3721 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3781 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3841 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3901 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3961 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4021 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4081 gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt
     4141 tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa
     4201 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga
     4261 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg
     4321 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc
     4381 gaggctgatc agcgggttta aacgggccct gctagagatt ttccacactg actaaaaggg
     4441 tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac
     4501 tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
     4561 ggctcagatc tggtctaacc agagagaccc agtacagtcc ggatgcagct ctcgggccat
     4621 gtgatgaaat gctaggcggc tgtcaaacct ccactctaat acttctctct ccgggtcatc
     4681 catcccatgc aggctcacag ggtgtaacaa gcgggtgttc tctccttcat tggcttcttc
     4741 taccttctct tgctcaactg gtactagctt gtagcaccat ccaaaggtca gtggatatct
     4801 gatccctggc cctggtgtgt agttctgcca atcagggaag tagccttgtg tgtggtagat
     4861 ccacagatca aggatatctt gtcttcgttg ggagtgaatt agcccttcca gtcccccctt
     4921 ttcttttaaa aagtggctaa gatctacagc tgccttgtaa gtcattggtc ttaaaggtac
     4981 ctgaggtgtg actggaaaac ccacctcctc ctcctcttgt gcttctagcc aggcacaatc
     5041 agcattggta gctgctgtat tgctacttgt gattgctcca tgtttttcta ggtctcgagg
     5101 tcgacggtat cgatgcgggg aggcggccca aagggagatc cgactcgtct gagggcgaag
     5161 gcgaagacgc ggaagaggcc gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg
     5221 attgagggcc gaagggacgt agcagaagga cgtcccgcgc agaatccagg tggcaacaca
     5281 ggcgagcagc caaggaaagg acgatgattt ccccgacaac accacggaat tgtcagtgcc
     5341 caacagccga gcccctgtcc agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc
     5401 aatagggagg gggaaagcga aagtcccgga aaggagctga caggtggtgg caatgcccca
     5461 accagtgggg gttgcgtcag caaacacagt gcacaccacg ccacgttgcc tgacaacggg
     5521 ccacaactcc tcataaagag acagcaacca ggatttatac aaggaggaga aaatgaaagc
     5581 catacgggaa gcaatagcat gatacaaagg cattaaagca gcgtatccac atagcgtaaa
     5641 aggagcaaca tagttaagaa taccagtcaa tctttcacaa attttgtaat ccagaggttg
     5701 attatcgata agcttgatat cgaattcata acttcgtata atgtatgcta tacgaagtta
     5761 tctattatca actttaggca ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg
     5821 gcacctcgac gtcggcggtg acggtgaagc cgagccgctc gtagaagggg aggttgcggg
     5881 gcgcggaggt ctccaggaag gcgggcaccc cggcgcgctc ggccgcctcc actccgggga
     5941 gcacgacggc gctgcccaga cccttgccct ggtggtcggg cgacacgccg acggtggcca
     6001 ggaaccacgc gggctccttg ggccggtgcg gcgccaggag gccttccatc tgttgctgcg
     6061 cggccagccg ggaaccgctc aactcggcca tgcgcgggcc gatctcggcg aacaccgccc
     6121 ccgcttcgac gctctccggc gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc
     6181 acaccttgcc gatgtcgagc ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc
     6241 gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg
     6301 cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct
     6361 tgtactcggt catcggtcca ggattctctt cgacatctcc ggcttgtttc agcagagaga
     6421 agtttgttgc gccggatccc ttgtacagct cgtccatgcc gccggtggag tggcggccct
     6481 cggcgcgttc gtactgttcc acgatggtgt agtcctcgtt gtgggaggtg atgtccaact
     6541 tgatgttgac gttgtaggcg ccgggcagct gcacgggctt cttggccttg taggtggtct
     6601 tgacctcagc gtcgtagtgg ccgccgtcct tcagcttcag cctctgcttg atctcgccct
     6661 tcagggcgcc gtcctcgggg tacatccgct cggaggaggc ctcccagccc atggtcttct
     6721 tctgcattac ggggccgtcg gaggggaagt tggtgccgcg cagcttcacc ttgtagatga
     6781 actcgccgtc ctgcagggag gagtcctggg tcacggtcac cacgccgccg tcctcgaagt
     6841 tcatcacgcg ctcccacttg aagccctcgg ggaaggacag cttcaagtag tcggggatgt
     6901 cggcggggtg cttcacgtag gccttggagc cgtacatgaa ctgaggggac aggatgtccc
     6961 aggcgaaggg cagggggcca cccttggtca ccttcagctt ggcggtctgg gtgccctcgt
     7021 aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg gccgttcacg gagccctcca
     7081 tgtgcacctt gaagcgcatg aactccttga tgatggccat gttatcctcc tcgcccttgc
     7141 tcaccattct agagtcgacc tgcagcccaa gcttcgtcta acaaaaaagc caaaaacggc
     7201 cagaatttag cggacaattt actagtctaa cactgaaaat tacatattga cccaaatgat
     7261 tacatttcaa aaggtgccta aaaaacttca caaaacacac tcgccaaccc cgagcgcata
     7321 gttcaaaacc ggagcttcag ctacttaaga agataggtac ataaaaccga ccaaagaaac
     7381 tgacgcctca cttatccctc ccctcaccag aggtccggcg cctgtcgatt caggagagcc
     7441 taccctaggc ccgaaccctg cgtcctgcga cggagaaaag cctaccgcac acctaccggc
     7501 aggtggcccc accctgcatt ataagccaac agaacgggtg acgtcacgac acgacgaggg
     7561 cgcgcgctcc caaaggtacg ggtgcactgc ccaacggcac cgccataact gccgcccccg
     7621 caacagacga caaaccgagt tctccagtca gtgacaaact tcacgtcagg gtccccagat
     7681 ggtgccccag cccatctcac ccgaataaga gctttcccgc attagcgaag gcctcaagac
     7741 cttgggttct tgccgcccac catgcccccc accttgtttc aacgacctca cagcccgcct
     7801 cacaagcgtc ttccattcaa gactcgggaa cagccgccat tttgctgcgc tccccccaac
     7861 ccccagttca gggcaacctt gctcgcggac ccagactaca gcccttggcg gtctctccac
     7921 acgcttccgt cccaccgagc ggcccggcgg ccacgaaagc cccggccagc ccagcagccc
     7981 gctactcacc aagtgacgat cacagcgatc cacaaacaag aaccgcgacc caaatcccgg
     8041 ctgcgacgga actagctgtg ccacacccgg cgcgtcctta tataatcatc ggcgttcacc
     8101 gccccacgga gatccctccg cagaatcgcc gagaagggac tacttttcct cgcctgttcc
     8161 gctctctgga aagaaaacca gtgccctaga gtcacccaag tcccgtccta aaatgtcctt
     8221 ctgctgatac tggggttcta aggccgagtc ttatgagcag cgggccgctg tcctgagcgt
     8281 ccgggcggaa ggatcaggac gctcgctgcg cccttcgtct gacgtggcag cgctcgccgt
     8341 gaggaggggg gcgcccgcgg gaggcgccaa aacccggcgc ggaggccaga tctggagccg
     8401 acacgggtta atgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg
     8461 gcgaattggg taccgagggc ctatttccca tgattccttc atatttgcat atacgataca
     8521 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
     8581 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     8641 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     8701 tcttgtggaa aggacgaaac accggagacg gttgtaaatg agcacacaaa atacacatgc
     8761 taaaatatta tattctatga cctttataaa atcaaccaaa atcttctttt taataacttt
     8821 agtatcaata attagaattt ttatgttcct ttttgcaaac ttttaataaa aatgagcaaa
     8881 ataaaaaaac gctagtttta gtaactcgcg ttgttttctt cacctttaat aatagctact
     8941 ccaccacttg ttcctaagcg gtcagctcct gcttcaatca ttttttgagc atcttcaaat
     9001 gttctaactc caccagctgc tttaactaaa gcattgtctt taacaactga cttcattagt
     9061 ttaacatctt caaatgttgc acctgatttt gaaaatcctg ttgatgtttt aacaaattct
     9121 aatccagctt caacagctat ttcacaagct ttcatgattt cttcttttgt taataaacaa
     9181 ttttccataa tacatttaac aacatgtgat ccagctgctt tttttacagc tttcatgtct
     9241 tctaaaacta attcataatt tttgtctttt aatgcaccaa tatttaatac catatcaatt
     9301 tctgttgcac catctttaat tgcttcagaa acttcgaatg cttttgtagc tgttgtgcat
     9361 gcacctagag gaaaacctac aacatttgtt attcctacat ttgtgccttt taataattct
     9421 ttacaatagc ttgttcaata tgaattaaca caaactgttg caaaatcaaa ttcaattgct
     9481 tcatcacata attgtttaat ttcagctttc gtagcatctt gttttaataa tgtgtgatct
     9541 atatatttgt ttagtttcat tttttctcct atatattcat ttttaatttt aattctttaa
     9601 taatttcgtc tactttaact ttagcgtttt gaacagattc accaacacct ataaaataaa
     9661 tttttagttt aggttcagtt ccacttgggc gaacagcaaa tcatgactta tcttctaaat
     9721 aaaattttag taagtcttgt cctggcatat tatacattcc atcgatgtag tcttcaacat
     9781 taacaacttt aagtccagca atttgagtta agggtgttgc tctcaatgat ttcattaatg
     9841 gttcaatttt taatttcttt tcttctggtt taaaattcaa gtttaaagtg aaagtgtaat
     9901 atgcacccat ttctttaaat aaatcttcta aatagtctac taatgtttta ttttgttttt
     9961 tataaaatca agcagcctct gctattaata tagaagcttg tattccatct ttatctctag
    10021 ctgagtcatc aattacatat ccataacttt cttcataagc aaaaacaaaa tttaatccgt
    10081 tatcttcttc tttagcaatt tctctaccca ttcatttaaa tccagttaaa gtttttacaa
    10141 tattaactcc atatttttca tgagcgattc tatcacccaa atcacttgtt acaaaacttg
    10201 aatatagagc cggatttttt ggaatgctat ttaagcgttt tagatttgat aattttcaat
    10261 caattaaaat tggtcctgtt tgatttccat ctaatcttac aaaatgacca tcatgtttta
    10321 ttgccattcc aaatctgtca gcatctgggt cattcataat aataatatct gcatcatgtt
    10381 taataccata ttcaagcggt atttttcatg caggatcaaa ttctggattt ggatttacaa
    10441 catttttaaa tgtttcatct tcaaatgcat gctcttcaac ctcaataacg ttatatcctg
    10501 attcacgtaa tatttttggg gtaaatttag ttcctgttcc attaactgcg ctaaaaataa
    10561 tttttaaatc ttttttagct tcttgctctt ttttgtacgt ctctgtttaa gagctatgct
    10621 ggaaacagca tagcaagttt aaataaggct agtccgttat caacttgaaa aagtggcacc
    10681 gagtcggtgc tttttttccg cggtggagct ccagcttttg ttccctttag tgagggttaa
    10741 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtattaacca aactggatct
    10801 ctgctgtccc tgtaataaac ccgaaaattt tgaatttttg taatttgttt ttgtaattct
    10861 ttagtttgta tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca
    10921 atcccccctt ttcttttaaa attgtggatg aatactgcca tttgtctgca gaattggcgc
    10981 acgcagtgcc gatccgttca ctaatcgaat ggatctgtct ctgtctctct ctccaccttc
    11041 ttcttctatt ccttcgggcc tgtcgggtcc cctcggggtt gggaggtggg tctgaaacga
    11101 taatggtgaa tatccctgcc taactctatt cactatagaa agtacagcaa aaactattct
    11161 taaacctacc aagcctccta ctatcattat gaataatttt atataccaca gccaatttgt
    11221 tatgttaaac caattccaca aacttgccca tttatctaat tccaataatt cttgttcatt
    11281 cttttcttgc tggttttgcg attcttcaat taaggagtgt attaagcttg tgtaattgtt
    11341 aatttctctg tcccactcca tccaggtcgt gtgattccaa atctgttcca gagatttatt
    11401 actccaacta gcattccaag gcacagcagt ggtgcaaatg agttttccag agcaacccca
    11461 aatccccagg agctgttgat cctttaggta tctttccaca gccaggattc ttgcctggag
    11521 ctgcttgatg ccccagactg tgagttgcaa cagatgctgt tgcgcctcaa tagccctcag
    11581 caaattgttc tgctgctgca ctataccaga caataattgt ctggcctgta ccgtcagcgt
    11641 cattgacgct gcgcccatag tgcttcctgc tgctcccaag aacccaagga acaaagctcc
    11701 tattcccact gctctttttt ctctctgcac cactcttctc tttgccttgg tgggtgctac
    11761 tcctaatggt tcaattttta ctactttata tttatataat tcacttctcc aattgtccct
    11821 catatctcct cctccaggtc tgaagatcag cggccgcttg ctgtgcggtg gtcttacttt
    11881 tgttttgctc ttcctctatc ttgtctaaag cttccttggt gtcttttatc tctatccttt
    11941 gatgcacaca atagagggtt gctactgtat tatataatga tctaagttct tctgatcctg
    12001 tctgaaggga tggttgtagc tgtcccagta tttgtctaca gccttctgat gtttctaaca
    12061 ggccaggatt aactgcgaat cgttctagct ccctgcttgc ccatactata tgttttaatt
    12121 tatatttttt ctttccccct ggccttaacc gaattttttc ccatcgcgat ctaattctcc
    12181 cccgcttaat actgacgctc tcgcacccat ctctctcctt ctagcctccg ctagtcaaaa
    12241 tttttggcgt actcaccagt cgccgcccct cgcctcttgc cgtgcgcgct tcagcaagcc
    12301 gagtcctgcg tcgagagagc tcctctggtt tccctttcgc tttcaagtcc ctgttcgggc
    12361 gccactgcta gagattttcc acactgacta aaagggtctg agggatctct agttaccaga
    12421 gtcacacaac agacgggcac acactacttg aagcactcaa ggcaagcttt attgaggctt
    12481 aagcagtggg ttccctagtt agccagagag ctcccaggct cagatctggt ctaaccagag
    12541 agacccagta caggcaaaac gcgctgctta tatagacctc ccaccgtaca cgcctaccgc
    12601 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
    12661 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
    12721 accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga
    12781 ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat
    12841 gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca
    12901 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
    12961 aagtccctat tggcgttact atggga
//
