LOCUS       sequence-318821-       10951 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of HBG site 1 sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10951)
  AUTHORS   Zhang X, Zhu B, Chen L, Xie L, Yu W, Wang Y, Li L, Yin S, Yang L, Hu
            H, Han H, Li Y, Wang L, Chen G, Ma X, Geng H, Huang W, Pang X, Yang 
            Z, Wu Y, Siwko S, Kurita R, Nakamura Y, Yang L, Liu M, Li D
  TITLE     Dual base editor catalyzes both cytosine and adenine base 
            conversions in human cells.
  JOURNAL   Nat Biotechnol. 2020 Jun 1. pii: 10.1038/s41587-020-0527-y. doi: 
            10.1038/s41587-020-0527-y.
  PUBMED    32483363
REFERENCE   2  (bases 1 to 10951)
  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 
            Biotechnol. 2020 Jun 1. pii: 10.1038/s41587-020-0527-y. doi: 
            10.1038/s41587-020-0527-y."
COMMENT     Sequence Label: Lenti U6-HBG site1-EF1alpha-UGI-P2A-GFP
FEATURES             Location/Qualifiers
     source          1..10951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..28)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        13..342
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      193..328
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     255..274
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        390..437
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             456..830
                     /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    960..1081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(997..1016)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1051..1070
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1130..1146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1130..1146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1143..1165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1154..1170
                     /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(1178..1184,1185..1202,1203..1208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1223..1244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1361..1378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1532..2120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1612..1631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2291..3082,3083..3151))
                     /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     2914..2933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3152..3256)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3331..3350)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3522..3901
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3903..4101
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4056..4076
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4119..4299
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4346..4471
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4964..5197
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5382..5426
                     /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    5724..5841
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5886..6134
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5886..5906
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6057..6076
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6155..6230
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6256..7434
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6487..7425
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7382..7402
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             7494..7514
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7521..7586
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7599..7847
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7860..7880
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7881..7934
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(7935..7937,7938..8648)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(7935..7937,7938..8648)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(7977..7998)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8238..8257)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8585..8606
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8658..8714
                     /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             8715..9311
                     /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     9208..9228
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9327..9915
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9380..9400)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9798..9809)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9987..10220
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10246..10263)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10252..10476
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10522..10950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10609..10628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10819..10840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
       61 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
      121 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
      181 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
      241 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
      301 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
      361 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
      421 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
      481 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
      541 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
      601 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
      661 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
      721 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
      781 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
      841 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
      901 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
      961 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     1021 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     1081 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     1141 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1201 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1261 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1321 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1381 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1441 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1501 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1561 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1621 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1681 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1741 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1801 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1861 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1921 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1981 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2041 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2101 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2161 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2221 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2281 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     2341 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     2401 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     2461 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     2521 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     2581 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     2641 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     2701 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     2761 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     2821 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     2881 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     2941 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     3001 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     3061 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     3121 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     3181 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     3241 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     3301 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3361 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     3421 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     3481 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
     3541 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     3601 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     3661 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     3721 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     3781 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     3841 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     3901 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     3961 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     4021 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     4081 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
     4141 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     4201 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     4261 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     4321 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     4381 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     4441 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
     4501 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
     4561 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
     4621 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     4681 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     4741 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     4801 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     4861 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     4921 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     4981 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     5041 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     5101 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     5161 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     5221 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     5281 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     5341 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     5401 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     5461 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     5521 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     5581 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     5641 acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca attctgcaga
     5701 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
     5761 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
     5821 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggtta
     5881 attaagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     5941 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     6001 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     6061 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     6121 aaggacgaaa caccgttgac caatagcctt gacagtttta gagctagaaa tagcaagtta
     6181 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttgat
     6241 atccatcaca ctggcggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     6301 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
     6361 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     6421 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     6481 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
     6541 tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg
     6601 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
     6661 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
     6721 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
     6781 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
     6841 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
     6901 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
     6961 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
     7021 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
     7081 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     7141 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     7201 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     7261 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     7321 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     7381 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaaatacg
     7441 actcactata gggagaccca agctggctag ttaagcttgg taccgccacc atgcccaaga
     7501 agaagaggaa agtctcgagc gactacaaag accatgacgg tgattataaa gatcatgaca
     7561 tcgattacaa ggatgacgat gacaagtctg gtggttctac taatctgtca gatattattg
     7621 aaaaggagac cggtaagcaa ctggttatcc aggaatccat cctcatgctc ccagaggagg
     7681 tggaagaagt cattgggaac aagccggaaa gcgatatact cgtgcacacc gcctacgacg
     7741 agagcaccga cgagaatgtc atgcttctga ctagcgacgc ccctgaatac aagccttggg
     7801 ctctggtcat acaggatagc aacggtgaga acaagattaa gatgctctct ggtggttctc
     7861 ccaagaagaa gaggaaagtc gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     7921 agaatcctgg cccagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     7981 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     8041 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     8101 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     8161 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     8221 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     8281 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     8341 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     8401 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     8461 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     8521 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     8581 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     8641 tgtacaaggg atccggcgca acaaacttct ctctgctgaa acaagccgga gatgtcgaag
     8701 agaatcctgg accgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     8761 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     8821 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     8881 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     8941 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     9001 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     9061 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca
     9121 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     9181 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     9241 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     9301 ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat
     9361 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     9421 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     9481 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     9541 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     9601 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     9661 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     9721 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt
     9781 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     9841 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     9901 gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag
     9961 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
    10021 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
    10081 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
    10141 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
    10201 agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt
    10261 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
    10321 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
    10381 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
    10441 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
    10501 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
    10561 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
    10621 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
    10681 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
    10741 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
    10801 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
    10861 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
    10921 agctgattta acaaaaattt aacgcgaatt a
//
