LOCUS       sequence-335505-        9841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2nd generation lentiviral transfer plasmid encoding a U6 promoter 
            expressing a spyCas9 sgRNA (no spacer, BsmBI cutsites) and an EF1-a 
            promoter expressing mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9841)
  AUTHORS   Hamilton JR, Tsuchida CA, Nguyen DN, Shy BR, McGarrigle ER, Sandoval
            Espinoza CR, Carr D, Blaeschke F, Marson A, Doudna JA
  TITLE     Targeted delivery of CRISPR-Cas9 and transgenes enables complex 
            immune cell engineering.
  JOURNAL   Cell Rep. 2021 Jun 1;35(9):109207. doi: 
            10.1016/j.celrep.2021.109207.
  PUBMED    34077734
REFERENCE   2  (bases 1 to 9841)
  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: "2021-06-1"; volume: "35(9):109207. doi"; pages: " 
            10.1016/j.celrep.2021.109207"
COMMENT     Sequence Label: pJRH051
FEATURES             Location/Qualifiers
     source          1..9841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2371..2619
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2371..2391
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2542..2561
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        2775..3952
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3006..3943
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3900..3920
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3970..3979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4702..5290
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4755..4775)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5173..5184)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5362..5595
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5621..5638)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5627..5851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5897..6325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5984..6003)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6194..6215
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6334..6354)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6339..6668
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6519..6654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6581..6600
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6716..6763
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6782..7156
                     /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    7286..7407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7323..7342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7377..7396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7456..7472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7456..7472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7469..7491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7480..7496
                     /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(7504..7510,7511..7528,7529..7534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7549..7570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7687..7704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7858..8446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7938..7957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8617..9408,9409..9477))
                     /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     9240..9259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9478..9582)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9657..9676)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat
     2401 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2461 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2521 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2581 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gagacggagg acgacgaacg
     2641 tctctgttta agagctatgc tggaaacagc atagcaagtt taaataaggc tagtccgtta
     2701 tcaacttgaa aaagtggcac cgagtcggtg ctttttttga attcgctagc taggtcttga
     2761 aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     2821 ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg
     2881 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     2941 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     3001 cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt
     3061 gcgtgccttg aattacttcc actggctgca gtacgtgatt cttgatcccg agcttcgggt
     3121 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
     3181 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
     3241 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
     3301 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
     3361 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag
     3421 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
     3481 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
     3541 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
     3601 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
     3661 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc
     3721 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
     3781 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
     3841 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
     3901 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gaaccggttc
     3961 tagagcgctg ccaccatggt aagtaagggc gaagaggata atatggcttc tttgcccgcc
     4021 acacacgaac tgcacatatt cggatcaata aacggggtgg atttcgatat ggtcggccag
     4081 ggaacaggaa accctaatga cggctacgaa gagctcaatc tcaaatctac caaaggggac
     4141 ttacaattct ctccatggat cctagtgcca catattggat atggcttcca tcaatacctt
     4201 ccatatccag acggtatgag ccccttccag gctgcaatgg tggacgggtc tggatatcag
     4261 gttcacagaa caatgcaatt tgaagacggt gcatccctta cagttaatta caggtacaca
     4321 tacgaaggaa gtcacatcaa aggggaggcg caggtgaagg gcaccgggtt cccagctgat
     4381 ggacctgtaa tgacaaattc actcacagcc gccgattggt gcaggtcaaa gaagacctat
     4441 cctaatgaca aaacaattat atccacattt aaatggtctt ataccacagg aaatggaaag
     4501 agatacagat ctacagctag aacaacttac acatttgcta aaccaatggc agccaactac
     4561 cttaagaatc aaccaatgta tgtcttcaga aagactgaac taaaacacag taaaacagaa
     4621 cttaatttca aagagtggca gaaggccttc acagacgtta tgggtatgga tgaactgtac
     4681 aagtaaacgc gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact
     4741 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     4801 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     4861 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     4921 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     4981 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     5041 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     5101 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     5161 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     5221 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     5281 gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact
     5341 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     5401 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     5461 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     5521 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     5581 ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     5641 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     5701 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     5761 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     5821 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     5881 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5941 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     6001 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6061 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6121 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6181 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6241 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6301 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     6361 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6421 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6481 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6541 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6601 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6661 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag cacgtgttga
     6721 caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac
     6781 catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt
     6841 cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg acttcgccgg
     6901 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga
     6961 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga
     7021 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca
     7081 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc
     7141 cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct tctatgaaag
     7201 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7261 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7321 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7381 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7441 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7501 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7561 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7621 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7681 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7741 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7801 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7861 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7921 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7981 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8041 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8101 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8161 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8221 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8281 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8341 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8401 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8461 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8521 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8581 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8641 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8701 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8761 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8821 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8881 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8941 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9001 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9061 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9121 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9181 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9241 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9301 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9361 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9421 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9481 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9541 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9601 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     9661 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     9721 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     9781 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     9841 t
//
