LOCUS       sequence-330251-       10592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of sgRNA with hygromycin resistance gene and 
            EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10592)
  AUTHORS   Ren X, Wang M, Li B, Jamieson K, Zheng L, Jones IR, Li B, Takagi MA,
            Lee J, Maliskova L, Tam TW, Yu M, Hu R, Lee L, Abnousi A, Li G, Li 
            Y, Hu M, Ren B, Wang W, Shen Y
  TITLE     Parallel characterization of cis-regulatory elements for multiple 
            genes using CRISPRpath.
  JOURNAL   Sci Adv. 2021 Sep 17;7(38):eabi4360. doi: 10.1126/sciadv.abi4360. 
            Epub 2021 Sep 15.
  PUBMED    34524848
REFERENCE   2  (bases 1 to 10592)
  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: "Sci Adv."; 
            date: "2021-09-17"; volume: "7(38):eabi4360. doi"; pages: " 
            10.1126/sciadv.abi4360. Epub 2021 Sep 15"
COMMENT     Sequence Label: lentiCRISPR-v2-HygR-EGFP
FEATURES             Location/Qualifiers
     source          1..10592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1990..2238
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1990..2010
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2161..2180
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        2996..3207
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3227..4246
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     "
     CDS             4265..4318
                     /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"
     regulatory      4325..4334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4331..4333,4334..4336,4337..5050)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4331..4333,4334..4336,4337..5047)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4376..4397)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4637..4656)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4984..5005
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5066..5654
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5119..5139)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5537..5548)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5726..5959
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5985..6002)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5991..6215
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6261..6689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6348..6367)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6558..6579
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6698..6718)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6703..7032
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6883..7018
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6945..6964
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7080..7127
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7146..7520
                     /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    7650..7771
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7687..7706)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7741..7760
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7820..7836)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7820..7836)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7833..7855)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7844..7860
                     /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(7868..7874,7875..7892,7893..7898))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7913..7934
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8051..8068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8222..8810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8302..8321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8981..9772,9773..9841))
                     /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     9604..9623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9842..9946)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10021..10040)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10212..10591
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     2041 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
     2101 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     2161 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     2221 tggaaaggac gaaacaccgg agacggttgt aaatgagcac acaaaataca catgctaaaa
     2281 tattatattc tatgaccttt ataaaatcaa ccaaaatctt ctttttaata actttagtat
     2341 caataattag aatttttatg ttcctttttg caaactttta ataaaaatga gcaaaataaa
     2401 aaaacgctag ttttagtaac tcgcgttgtt ttcttcacct ttaataatag ctactccacc
     2461 acttgttcct aagcggtcag ctcctgcttc aatcattttt tgagcatctt caaatgttct
     2521 aactccacca gctgctttaa ctaaagcatt gtctttaaca actgacttca ttagtttaac
     2581 atcttcaaat gttgcacctg attttgaaaa tcctgttgat gttttaacaa attctaatcc
     2641 agcttcaaca gctatttcac aagctttcat gatttcttct tttgttaata aacaattttc
     2701 cataatacat ttaacaacat gtgatccagc tgcttttttt acagctttca tgtcttctaa
     2761 aactaattca taatttttgt cttttaatgc accaatattt aataccatat caatttctgt
     2821 tgcaccatct ttaattgcta cgtctctgtt tcagagctat gctggaaaca gcatagcaag
     2881 ttgaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
     2941 gaattcgcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca
     3001 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt
     3061 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     3121 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     3181 cgcaacgggt ttgccgccag aacacaggac cggttctaga gccaccatga aaaagcctga
     3241 actcaccgct acctctgtcg agaagtttct gatcgaaaag ttcgacagcg tgtccgacct
     3301 gatgcagctc tccgagggcg aagaatctcg ggctttcagc ttcgatgtgg gagggcgtgg
     3361 atatgtcctg cgggtgaata gctgcgccga tggtttctac aaagatcgct atgtttatcg
     3421 gcactttgca tccgccgctc tccctattcc cgaagtgctt gacattgggg agttcagcga
     3481 gagcctgacc tattgcatct cccgccgtgc acagggtgtc accttgcaag acctgcctga
     3541 aaccgaactg cccgctgttc tccagcccgt cgccgaggcc atggatgcca tcgctgccgc
     3601 cgatcttagc cagaccagcg ggttcggccc attcggacct caaggaatcg gtcaatacac
     3661 tacatggcgc gatttcatct gcgctattgc tgatccccat gtgtatcact ggcaaactgt
     3721 gatggacgac accgtcagtg cctccgtcgc ccaggctctc gatgagctga tgctttgggc
     3781 cgaggactgc cccgaagtcc ggcacctcgt gcacgccgat ttcggctcca acaatgtcct
     3841 gaccgacaat ggccgcataa cagccgtcat tgactggagc gaggccatgt tcggggattc
     3901 ccaatacgag gtcgccaaca tcttcttctg gaggccctgg ttggcttgta tggagcagca
     3961 gacccgctac ttcgagcgga ggcatcccga gcttgcagga tctcctcggc tccgggctta
     4021 tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga
     4081 tgcagcttgg gctcagggtc gctgcgacgc aatcgtccgg tccggagccg ggactgtcgg
     4141 gcgtacacaa atcgcccgca gaagcgctgc cgtctggacc gatggctgtg tggaagtgct
     4201 cgccgatagt ggaaacagac gccccagcac tcgtcctagg gcaaaggatc tgcagggcag
     4261 tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccaag
     4321 cgctgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4381 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     4441 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     4501 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     4561 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     4621 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     4681 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     4741 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     4801 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     4861 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     4921 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     4981 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     5041 gtacaagtaa acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
     5101 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5161 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     5221 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     5281 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     5341 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     5401 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     5461 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     5521 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5581 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     5641 ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
     5701 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
     5761 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5821 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5881 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5941 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     6001 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     6061 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     6121 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     6181 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     6241 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     6301 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     6361 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     6421 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6481 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     6541 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     6601 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6661 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     6721 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     6781 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6841 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     6901 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     6961 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     7021 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     7081 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     7141 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
     7201 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
     7261 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
     7321 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
     7381 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
     7441 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
     7501 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
     7561 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     7621 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     7681 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     7741 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     7801 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7861 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7921 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7981 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     8041 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     8101 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     8161 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     8221 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     8281 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     8341 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     8401 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     8461 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     8521 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     8581 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     8641 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     8701 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     8761 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     8821 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8881 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8941 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     9001 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     9061 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     9121 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     9181 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     9241 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     9301 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     9361 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     9421 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     9481 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     9541 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     9601 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     9661 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     9721 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     9781 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9841 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9901 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9961 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
    10021 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
    10081 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
    10141 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
    10201 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
    10261 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
    10321 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
    10381 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
    10441 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
    10501 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
    10561 catctacgta ttagtcatcg ctattaccat gg
//
