LOCUS       sequence-245287-       10168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of sgRNA with GFP and neomycin resistance 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10168)
  AUTHORS   Tarumoto Y, Lin S, Wang J, Milazzo JP, Xu Y, Lu B, Yang Z, Wei Y, 
            Polyanskaya S, Wunderlich M, Gray NS, Stegmaier K, Vakoc CR
  TITLE     Salt-Inducible Kinase inhibition suppresses acute myeloid leukemia 
            progression in vivo.
  JOURNAL   Blood. 2019 Nov 1. pii: 422697. doi: 10.1182/blood.2019001576.
  PUBMED    31697837
REFERENCE   2  (bases 1 to 10168)
  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: "Blood. 2019
            Nov 1. pii: 422697. doi: 10.1182/blood.2019001576."
COMMENT     Sequence Label: LRG2.1_Neo
FEATURES             Location/Qualifiers
     source          1..10168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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"
     promoter        1717..1965
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1717..1737
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1888..1907
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    3991..4108
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4177..4388
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4408..4417
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4417..4440
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(4441..4443,4444..4446,4447..5157)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4441..4443,4444..4446,4447..5157)
                     /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(4486..4507)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4747..4766)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5094..5115
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5167..5223
                     /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             5224..6018
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5278..5297)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5888..5907
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    6025..6613
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6078..6098)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6496..6507)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6685..6918
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6996..7117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7033..7052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7087..7106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7157..7292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7312..7331)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7313..7331)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7341..7358)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7341..7357)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7350..7372)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7499..7954
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7586..7605)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7796..7817
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7980..8084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8085..8153,8154..8945)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8303..8322)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9116..9704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9605..9624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9858..9875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9992..10013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10028..10033,10034..10051,10052..10058)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10066..10082
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     10071..10093
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10090..10106
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10090..10106
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10125..10145
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10127..10145
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct
     1741 tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta
     1801 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     1861 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     1921 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaga cggttgtaaa
     1981 tgagcacaca aaatacacat gctaaaatat tatattctat gacctttata aaatcaacca
     2041 aaatcttctt tttaataact ttagtatcaa taattagaat ttttatgttc ctttttgcaa
     2101 acttttaata aaaatgagca aaataaaaaa acgctagttt tagtaactcg cgttgttttc
     2161 ttcaccttta ataatagcta ctccaccact tgttcctaag cggtcagctc ctgcttcaat
     2221 cattttttga gcatcttcaa atgttctaac tccaccagct gctttaacta aagcattgtc
     2281 tttaacaact gacttcatta gtttaacatc ttcaaatgtt gcacctgatt ttgaaaatcc
     2341 tgttgatgtt ttaacaaatt ctaatccagc ttcaacagct atttcacaag ctttcatgat
     2401 ttcttctttt gttaataaac aattttccat aatacattta acaacatgtg atccagctgc
     2461 tttttttaca gctttcatgt cttctaaaac taattcataa tttttgtctt ttaatgcacc
     2521 aatatttaat accatatcaa tttctgttgc accatcttta attgcttcag aaacttcgaa
     2581 tgcttttgta gctgttgtgc atgcacctag aggaaaacct acaacatttg ttattcctac
     2641 atttgtgcct tttaataatt ctttacaata gcttgttcaa tatgaattaa cacaaactgt
     2701 tgcaaaatca aattcaattg cttcatcaca taattgttta atttcagctt tcgtagcatc
     2761 ttgttttaat aatgtgtgat ctatatattt gtttagtttc attttttctc ctatatattc
     2821 atttttaatt ttaattcttt aataatttcg tctactttaa ctttagcgtt ttgaacagat
     2881 tcaccaacac ctataaaata aatttttagt ttaggttcag ttccacttgg gcgaacagca
     2941 aatcatgact tatcttctaa ataaaatttt agtaagtctt gtcctggcat attatacatt
     3001 ccatcgatgt agtcttcaac attaacaact ttaagtccag caatttgagt taagggtgtt
     3061 gctctcaatg atttcattaa tggttcaatt tttaatttct tttcttctgg tttaaaattc
     3121 aagtttaaag tgaaagtgta atatgcaccc atttctttaa ataaatcttc taaatagtct
     3181 actaatgttt tattttgttt tttataaaat caagcagcct ctgctattaa tatagaagct
     3241 tgtattccat ctttatctct agctgagtca tcaattacat atccataact ttcttcataa
     3301 gcaaaaacaa aatttaatcc gttatcttct tctttagcaa tttctctacc cattcattta
     3361 aatccagtta aagtttttac aatattaact ccatattttt catgagcgat tctatcaccc
     3421 aaatcacttg ttacaaaact tgaatataga gccggatttt ttggaatgct atttaagcgt
     3481 tttagatttg ataattttca atcaattaaa attggtcctg tttgatttcc atctaatctt
     3541 acaaaatgac catcatgttt tattgccatt ccaaatctgt cagcatctgg gtcattcata
     3601 ataataatat ctgcatcatg tttaatacca tattcaagcg gtatttttca tgcaggatca
     3661 aattctggat ttggatttac aacattttta aatgtttcat cttcaaatgc atgctcttca
     3721 acctcaataa cgttatatcc tgattcacgt aatatttttg gggtaaattt agttcctgtt
     3781 ccattaactg cgctaaaaat aatttttaaa tcttttttag cttcttgctc ttttttgtac
     3841 gtctctgttt aagagctatg ctggaaacag catagcaagt ttaaataagg ctagtccgtt
     3901 atcaactttg ctggaaacag caaagtggca ccgagtcggt gctttttttg aattctagat
     3961 cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     4021 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     4081 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact
     4141 ttggcgccgg ctcgagtggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     4201 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
     4261 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     4321 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     4381 gaacacaggt gtcgtgacgc gggatccgcc accatggatt acaaagacga tgacgataag
     4441 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4501 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     4561 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4621 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     4681 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     4741 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     4801 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4861 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     4921 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     4981 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     5041 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     5101 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc
     5161 agcggcgcca ccaacttcag cctgctgaag caggccggcg acgtggagga gaaccccggc
     5221 cccatgcttg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5281 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5341 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5401 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5461 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5521 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5581 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5641 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5701 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5761 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5821 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5881 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5941 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6001 cttgacgagt tcttctgagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     6061 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     6121 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     6181 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     6241 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     6301 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     6361 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     6421 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     6481 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6541 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6601 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     6661 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     6721 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6781 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6841 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6901 tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat
     6961 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     7021 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7081 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     7141 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     7201 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     7261 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     7321 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     7381 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     7441 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     7501 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     7561 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     7621 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     7681 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     7741 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7801 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7861 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7921 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     7981 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     8041 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     8101 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     8161 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     8221 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     8281 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     8341 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8401 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8461 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8521 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     8581 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     8641 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8701 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8761 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     8821 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8881 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8941 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     9001 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     9061 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9121 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9181 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9241 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     9301 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     9361 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     9421 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     9481 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     9541 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     9601 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     9661 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     9721 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     9781 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     9841 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     9901 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     9961 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
    10021 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
    10081 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
    10141 aaagggaaca aaagctggag ctgcaagc
//
