LOCUS       sequence-89669-e       12096 bp    DNA     circular SYN 14-APR-2025
DEFINITION  GATEWAY-compatible lentiviral conditional RNAi delivery vector; 
            SFFV-driven expression of TetR and eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12096)
  AUTHORS   Sigl R, Ploner C, Shivalingaiah G, Kofler R, Geley S
  TITLE     Development of a Multipurpose GATEWAY-Based Lentiviral 
            Tetracycline-Regulated Conditional RNAi System (GLTR).
  JOURNAL   PLoS One. 2014 May 19;9(5):e97764. doi: 
            10.1371/journal.pone.0097764. eCollection 2014.
  PUBMED    24841113
REFERENCE   2  (bases 1 to 12096)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2014-05-19"; volume: "9(5):e97764. doi"; pages: " 
            10.1371/journal.pone.0097764. eCollection 2014"
COMMENT     Sequence Label: pGLTR-X-GFP
FEATURES             Location/Qualifiers
     source          1..12096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        98..505
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      514..523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             520..1140
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MARLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(593..612)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             1141..1161
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1174..1227
                     /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      1243..1252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1249..1251,1252..1254,1255..1968)
                     /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(1249..1251,1252..1254,1255..1965)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1294..1315)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1555..1574)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1902..1923
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2026..2614
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2079..2099)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2497..2508)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2617..2633)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2618..2634)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             2724..2957
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(2982..3000)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2983..3000)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3290..3309)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3522..3541)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3641..3663
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3701..3719)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3787..3891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3892..3960,3961..4752)
                     /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(4110..4129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4923..5511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5412..5431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5665..5682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5752..5772)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5757..6086
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5937..6072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          7220..7285
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             7415..7435
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7860..7994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7897..7916)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7951..7970
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8163..8796
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    8843..8968
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    9460..9693
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9878..9922
                     /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    10207..10324
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    10388..10512
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(10537..10542,10543..10560,10561..10567)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             10621..11280
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(10687..10706)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             11622..11927
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     11806..11825
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(11968..12092)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 aattcctgca gccccgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
       61 gaaagacccc acctgtaggt ttggcaagct agctgcagta acgccatttt gcaaggcatg
      121 gaaaaatacc aaaccaagaa tagagaagtt cagatcaagg gcgggtacat gaaaatagct
      181 aacgttgggc caaacaggat atctgcggtg agcagtttcg gccccggccc ggggccaaga
      241 acagatggtc accgcagttt cggccccggc ccgaggccaa gaacagatgg tccccagata
      301 tggcccaacc ctcagcagtt tcttaagacc catcagatgt ttccaggctc ccccaaggac
      361 ctgaaatgac cctgcgcctt atttgaatta accaatcagc ctgcttctcg cttctgttcg
      421 cgcgcttctg cttcccgagc tctataaaag agctcacaac ccctcactcg gcgcgccagt
      481 cctccgacag actgagtcgc ccggggggga tccgccacca tggctagatt agataaaagt
      541 aaagtgatta acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc
      601 cgtaaactcg cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat
      661 aagcgggctt tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt
      721 tgccctttag aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga
      781 tgtgctttac taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa
      841 aaacagtatg aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta
      901 gagaatgcat tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa
      961 gatcaagagc atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg
     1021 ccattattac gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta
     1081 ttcggccttg aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtggttcc
     1141 ccaaaaaaga agagaaaggt cgacggcggt ggtgaaggcc gcggcagcct gctgacctgc
     1201 ggcgatgtgg aagaaaaccc gggcccgggg gatccaccgg tcgccaccat ggtgagcaag
     1261 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1321 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1381 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1441 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1501 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1561 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1621 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1681 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1741 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1801 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1861 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1921 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac
     1981 tctagagtcg actgcagcat gaagcttgat atcaagctta tcgataatca acctctggat
     2041 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2101 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2161 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2221 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2281 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2341 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2401 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     2461 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2521 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     2581 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagggaat
     2641 taattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     2701 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     2761 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     2821 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     2881 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     2941 gtggaaaatc tctagcagca tctagaatta attccgtgta ttctatagtg tcacctaaat
     3001 cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa cgacaatatg
     3061 tacaagccta attgtgtagc atctggctta ctgaagcaga ccctatcatc tctctcgtaa
     3121 actgccgtca gagtcggttt ggttggacga accttctgag tttctggtaa cgccgtcccg
     3181 cacccggaaa tggtcagcga accaatcagc agggtcatcg ctagccagat cctctacgcc
     3241 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     3301 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     3361 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     3421 ccattccttg cggcggcggt gctcaacggc ctcaactact actgggctgc ttcctaatgc
     3481 aggagtcgca taagggagag cgtcgaatgg tgcactctca gtacaatctg ctctgatgcc
     3541 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3601 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3661 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     3721 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3781 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3841 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3901 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3961 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4021 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4081 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4141 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4201 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4261 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4321 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4381 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4441 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4501 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4561 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4621 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4681 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4741 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4801 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4861 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4921 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4981 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5041 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     5101 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5161 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5221 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5281 acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa
     5341 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5401 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5461 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5521 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5581 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5641 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5701 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg tggaatgtgt
     5761 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5821 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5881 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5941 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6001 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6061 tttttggagg cctaggcttt tgcaaaaagc ttggacacaa gacaggcttg cgagatatgt
     6121 ttgagaatac cactttatcc cgcgtcaggg agaggcagtg cgtaaaaaga cgcggactca
     6181 tgtgaaatac tggtttttag tgcgccagat ctctataatc tcgcgcaacc tattttcccc
     6241 tcgaacactt tttaagccgt agataaacag gctgggacac ttcacatgag cgaaaaatac
     6301 atcgtcacct gggacatgtt gcagatccat gcacgtaaac tcgcaagccg actgatgcct
     6361 tctgaacaat ggaaaggcat tattgccgta agccgtggcg gtctgtaccg ggtgcgttac
     6421 tggcgcgtga actgggtatt cgtcatgtcg ataccgtttg tatttccagc tacgatcacg
     6481 acaaccagcg cgagcttaaa gtgctgaaac gcgcagaagg cgatggcgaa ggcttcatcg
     6541 ttattgatga cctggtggat accggtggta ctgcggttgc gattcgtgaa atgtatccaa
     6601 aagcgcactt tgtcaccatc ttcgcaaaac cggctggtcg tccgctggtt gatgactatg
     6661 ttgttgatat cccgcaagat acctggattg aacagccgtg ggatatgggc gtcgtattcg
     6721 tcccgccaat ctccggtcgc taatcttttc aacgcctggc actgccgggc gttgttcttt
     6781 ttaacttcag gcgggttaca atagtttcca gtaagtattc tggaggctgc atccatgaca
     6841 caggcaaacc tgagcgaaac cctgttcaaa ccccgcttta aacatcctga aacctcgacg
     6901 ctagtccgcc gctttaatca cggcgcacaa ccgcctgtgc agtcggccct tgatggtaaa
     6961 accatccctc actggtatcg catgattaac cgtctgatgt ggatctggcg cggcattgac
     7021 ccacgcgaaa tcctcgacgt ccaggcacgt attgtgatga gcgatgccga acgtaccgac
     7081 gatgatttat acgatacggt gattggctac cgtggcggca actggattta tgagtgggcc
     7141 ccggatcttt gtgaaggaac cttacttctg tggtgtgaca taattggaca aactacctac
     7201 agagatttaa agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactact
     7261 gattctaatt gtttgtgtat tttagattcc aacctatgga actgatgaat gggagcagtg
     7321 gtggaatgcc tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga
     7381 tgaggctact gctgactctc aacattctac tcctccaaaa aagaagagaa aggtagaaga
     7441 ccccaaggac tttccttcag aattgctaag ttttttgagt catgctgtgt ttagtaatag
     7501 aactcttgct tgctttgcta tttacaccac aaaggaaaaa gctgcactgc tatacaagaa
     7561 aattatggaa aaatattctg taacctttat aagtaggcat aacagttata atcataacat
     7621 actgtttttt cttactccac acaggcatag agtgtctgct attaataact atgctcaaaa
     7681 attgtgtacc tttagctttt taatttgtaa aggggttaat aaggaatatt tgatgtatag
     7741 tgccttgact agagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     7801 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     7861 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     7921 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7981 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
     8041 cataccctat taccactgcc aattacctag tggtttcatt tactctaaac ctgtgattcc
     8101 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
     8161 gttggaaggg ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca
     8221 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     8281 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     8341 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     8401 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     8461 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     8521 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     8581 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     8641 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     8701 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     8761 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     8821 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     8881 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     8941 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     9001 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     9061 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     9121 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     9181 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     9241 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     9301 gcaagcnnng ctgatcttca gacctggagg aggagatnnn agggacaatt ggagaagtga
     9361 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     9421 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     9481 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     9541 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     9601 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     9661 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     9721 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     9781 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     9841 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     9901 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     9961 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
    10021 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
    10081 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
    10141 agacagatcc attcgattag tgaacggatc tcgacggtat cgccaaatgg cagtattcat
    10201 ccacaatttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
    10261 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
    10321 ttttcgggtt tattacaggg acagcagaga tccagtttgg atcgataagc ttgatatcga
    10381 attatcaaca agtttgtaca aaaaagctga acgagaaacg taaaatgata taaatatcaa
    10441 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat
    10501 ccagtcatat tggcggccgc attaggcacc ccaggcttta cactttatgc ttccggctcg
    10561 tataatgtgt ggattttgag ttaggatccg tcgagatttt caggagctaa ggaagctaaa
    10621 atggagaaaa aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa
    10681 cattttgagg catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat
    10741 attacggcct ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt
    10801 cacattcttg cccgcctgat gaatgctcat ccggaattcc gtatggcaat gaaagacggt
    10861 gagctggtga tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa
    10921 acgttttcat cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat
    10981 tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag
    11041 aatatgtttt tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg
    11101 gccaatatgg acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc
    11161 gacaaggtgc tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat
    11221 gtcggcagaa tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa
    11281 agatctggat ccggcttact aaaagccaga taacagtatg cgtatttgcg cgctgatttt
    11341 tgcggtataa gaatatatac tgatatgtat acccgaagta tgtcaaaaag aggtatgcta
    11401 tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc aaggcatata
    11461 tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg
    11521 tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc ggtttattga
    11581 aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt aaggtttaca
    11641 cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca
    11701 cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct
    11761 cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg
    11821 atatggccag tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg
    11881 aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc
    11941 ttatacacag ccagtctgca ggtcgaccat agtgactgga tatgttgtgt tttacagtat
    12001 tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac
    12061 gtttctcgtt cagctttctt gtacaaagtg gttgat
//
