LOCUS       sequence-342766-       10272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-ON miR-E (miR-30 variant)-based RNAi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10272)
  AUTHORS   Fellmann C, Hoffmann T, Sridhar V, Hopfgartner B, Muhar M, Roth M, 
            Lai DY, Barbosa IA, Kwon JS, Guan Y, Sinha N, Zuber J
  TITLE     An optimized microRNA backbone for effective single-copy RNAi.
  JOURNAL   Cell Rep. 2013 Dec 26;5(6):1704-13. doi: 
            10.1016/j.celrep.2013.11.020. Epub 2013 Dec 12.
  PUBMED    24332856
REFERENCE   2  (bases 1 to 10272)
  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"; doi: 
            "10.1016/j.celrep.2013.11.020"; journalName: "Cell Rep"; date: 
            "2013-12-26- 26"; volume: "5"; issue: "6"; pages: "1704-13"
COMMENT     Sequence Label: LT3GEPIR
FEATURES             Location/Qualifiers
     source          1..10272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..31
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    148..169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(184..189,190..207,208..214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    222..238
                     /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     227..249
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     246..262
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     246..262
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     281..301
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        283..301
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        329..555
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             556..736
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    783..908
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1401..1634
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1819..1863
                     /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    2130..2247
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2263..2641
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2271..2289
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2307..2325
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2343..2361
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2379..2397
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2415..2433
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2451..2469
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2487..2505
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2550..2574
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2665..2667,2668..2670,2671..3384)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2710..2731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2971..2990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3318..3339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           3578..3592
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        3771..4270
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3806..3829)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4203..4222
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4291..4890
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4291..4310)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4787..4807
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4951..5518
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4968..5520
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5112..5129)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5339..5358
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5532..6149,6150..6155,6156..6191,6192..6194,6195..6230,
                     6231..6233,6234..6269,6270..6278)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    6324..6912
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6377..6397)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6795..6806)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7119..7352
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7424..7545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7461..7480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7515..7534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7585..7720
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7740..7759)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7741..7759)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7769..7786)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7769..7785)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7778..7800)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7927..8382
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8014..8033)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8224..8245
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8408..8512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8513..8581,8582..9373)
                     /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(8731..8750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9544..10132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10033..10052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
       61 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
      121 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      181 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      241 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
      301 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
      361 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
      421 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
      481 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
      541 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
      601 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
      661 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
      721 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
      781 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
      841 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
      901 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
      961 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     1021 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     1081 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     1141 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     1201 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     1261 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     1321 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     1381 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     1441 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     1501 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     1561 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     1621 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     1681 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     1741 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     1801 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     1861 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     1921 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     1981 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
     2041 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
     2101 gtgaacggat ctcgacggta tcggttaact tttaaaagaa aaggggggat tggggggtac
     2161 agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa
     2221 aaacaaatta caaaaattca aaattttatc gattctagat tcgagtttac tccctatcag
     2281 tgatagagaa cgtatgaaga gtttactccc tatcagtgat agagaacgta tgcagacttt
     2341 actccctatc agtgatagag aacgtataag gagtttactc cctatcagtg atagagaacg
     2401 tatgaccagt ttactcccta tcagtgatag agaacgtatc tacagtttac tccctatcag
     2461 tgatagagaa cgtatatcca gtttactccc tatcagtgat agagaacgta taagctttag
     2521 gcgtgtacgg tgggcgccta taaaagcaga gctcgtttag tgaaccgtca gatcgcctgg
     2581 agcaattcca caacactttt gtcttatacc aactttccgt accacttcct accctcgtaa
     2641 agtcgagctt gcgttggatc caccatggtg agcaagggcg aggagctgtt caccggggtg
     2701 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtctggc
     2761 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2821 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2881 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2941 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3001 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3061 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3121 atcatggccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc
     3181 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3241 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3301 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3361 ggcatggacg agctgtacaa gtaatgacgt acggcggccg cctcgactag ggataacagg
     3421 gtaattgttt gaatgaggct tcagtacttt acagaatcgt tgcctgcaca tcttggaaac
     3481 acttgctggg attacttcga cttcttaacc caacagaagg ctcgagaagg tatattgctg
     3541 ttgacagtga gcgcaggaat tataatgctt atctatagtg aagccacaga tgtatagata
     3601 agcattataa ttcctatgcc tactgcctcg gacttcaagg ggctagaatt cgagcaatta
     3661 tcttgtttac taaaactgaa taccttgcta tctctttgat acatttttac aaagctgaat
     3721 taaaatggta taaattaaat cacttttttc aattgacgcg taattctacc gggtagggga
     3781 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     3841 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     3901 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     3961 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     4021 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     4081 ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg
     4141 gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
     4201 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
     4261 gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc
     4321 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     4381 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     4441 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     4501 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     4561 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     4621 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     4681 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     4741 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     4801 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     4861 tggtgcatga cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc
     4921 gaaaggagcg cacgacccca tgcatccaat tccgcccccc ccccctaacg ttactggccg
     4981 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     5041 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     5101 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     5161 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     5221 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     5281 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     5341 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     5401 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     5461 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     5521 tggccacaac catgagtaga ctggacaaga gcaaagtcat aaacggagct ctggaattac
     5581 tcaatggtgt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg
     5641 agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa
     5701 tcgagatgct ggacaggcat catacccact tctgccccct ggaaggcgag tcatggcaag
     5761 actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg
     5821 ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc
     5881 tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg
     5941 tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg
     6001 aaagagagac acctaccacc gattctatgc ccccacttct gagacaagca attgagctgt
     6061 tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc
     6121 tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact
     6181 tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg
     6241 ctcttgacga ttttgacctt gacatgctcc ccgggtaatc cggactcgac ctgcagccaa
     6301 gcttatcgaa acgcgtctgg aacaatcaac ctctggatta caaaatttgt gaaagattga
     6361 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     6421 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     6481 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     6541 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     6601 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6661 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     6721 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     6781 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     6841 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     6901 ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg gagcaatcac
     6961 aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac aagaggagga
     7021 ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     7081 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     7141 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     7201 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     7261 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     7321 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     7381 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     7441 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     7501 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     7561 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     7621 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     7681 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     7741 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     7801 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     7861 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     7921 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     7981 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     8041 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     8101 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     8161 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     8221 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     8281 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     8341 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     8401 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     8461 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     8521 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     8581 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     8641 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     8701 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     8761 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8821 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8881 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     8941 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     9001 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     9061 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     9121 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     9181 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     9241 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     9301 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     9361 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     9421 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     9481 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     9541 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     9601 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     9661 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     9721 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     9781 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     9841 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     9901 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     9961 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
    10021 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
    10081 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
    10141 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
    10201 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
    10261 tcgccgcagc cg
//
