LOCUS       sequence-339657-        9882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For cloning gRNA block.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9882)
  AUTHORS   Xiang X, Corsi GI, Anthon C, Qu K, Pan X, Liang X, Han P, Dong Z, 
            Liu L, Zhong J, Ma T, Wang J, Zhang X, Jiang H, Xu F, Liu X, Xu X, 
            Wang J, Yang H, Bolund L, Church GM, Lin L, Gorodkin J, Luo Y
  TITLE     Enhancing CRISPR-Cas9 gRNA efficiency prediction by data integration
            and deep learning.
  JOURNAL   Nat Commun. 2021 May 28;12(1):3238. doi: 10.1038/s41467-021-23576-0.
  PUBMED    34050182
REFERENCE   2  (bases 1 to 9882)
  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: "Nat 
            Commun."; date: "2021-05-28"; volume: "12(1):3238. doi"; pages: " 
            10.1038/s41467-021-23576-0"
COMMENT     Sequence Label: LentiU6-LacZ-GFP-Puro (BB)
FEATURES             Location/Qualifiers
     source          1..9882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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    1606..1723
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1774..2022
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1774..1794
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1945..1964
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             complement(1993..2307)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDVSGVSSFPQDI
                     "
     primer_bind     2157..2179
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2171..2188
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2172..2188
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2198..2217
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2198..2216
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2264..2284)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2264..2282)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2303..2319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2327..2343
                     /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(2351..2357,2358..2375,2376..2381))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2396..2417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2509..2720
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2739..2748
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2745..2747,2748..2750,2751..3461)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2745..2747,2748..2750,2751..3461)
                     /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(2790..2811)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3051..3070)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3398..3419
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3471..3527
                     /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             3528..4124
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     4021..4041
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4140..4728
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4193..4213)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4611..4622)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4800..5033
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5059..5076)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5065..5289
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5335..5763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5422..5441)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5632..5653
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5772..5792)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5777..6106
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5957..6092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6019..6038
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6154..6201
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6220..6594
                     /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    6724..6845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6761..6780)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6815..6834
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6894..6910)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6918..6934
                     /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(6942..6948,6949..6966,6967..6972))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6987..7008
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7125..7142)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7296..7884)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7376..7395)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8055..8846,8847..8915))
                     /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     8678..8697
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8916..9020)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9095..9114)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9286..9665
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9667..9865
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9820..9840
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatcggca
     1561 ctgcgtgcgc caattctgca gacaaatggc agtattcatc cacaatttta aaagaaaagg
     1621 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     1681 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     1741 cagcagagat ccagtttggt taattaaggt accgagggcc tatttcccat gattccttca
     1801 tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac
     1861 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
     1921 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
     1981 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggagacgt cccattcgcc
     2041 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     2101 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     2161 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     2221 attgggtacc gggccccccc tcgaggtcct ccagcttttg ttccctttag tgagggttaa
     2281 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2341 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2401 tgagctaact cacattaatt gcgttgcgct cactgcccgc ttcgtctctg tttgaattcg
     2461 ctagctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca
     2521 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc ggtgcctaga
     2581 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2641 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2701 ggtttgccgc cagaacacag gaccggttct agagcgctgc caccatggtg agcaagggcg
     2761 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2821 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2881 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2941 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3001 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3061 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3121 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3181 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     3241 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     3301 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     3361 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     3421 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggatccggc gcaacaaact
     3481 tctctctgct gaaacaagcc ggagatgtcg aagagaatcc tggaccgacc gagtacaagc
     3541 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     3601 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     3661 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     3721 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     3781 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     3841 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     3901 ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     3961 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     4021 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     4081 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaacgcgt taagtcgaca
     4141 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4201 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4261 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4321 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4381 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4441 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4501 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     4561 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4621 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4681 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag
     4741 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     4801 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     4861 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     4921 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     4981 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagggcccg
     5041 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     5101 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     5161 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     5221 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     5281 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     5341 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5401 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5461 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5521 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5581 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5641 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5701 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5761 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5821 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5881 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5941 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     6001 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     6061 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     6121 ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat
     6181 atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc
     6241 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
     6301 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
     6361 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
     6421 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
     6481 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
     6541 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg
     6601 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     6661 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     6721 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6781 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6841 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     6901 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     6961 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     7021 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     7081 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7141 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7201 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7261 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7321 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7381 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7441 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7501 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7561 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7621 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7681 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     7741 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7801 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7861 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7921 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7981 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     8041 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     8101 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     8161 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     8221 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     8281 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8341 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     8401 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8461 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8521 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8581 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8641 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     8701 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8761 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8821 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8881 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8941 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     9001 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     9061 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     9121 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     9181 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     9241 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     9301 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     9361 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     9421 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     9481 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     9541 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     9601 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     9661 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     9721 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     9781 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     9841 tacggtggga ggtctatata agcagcgcgt tttgcctgta ct
//
