LOCUS       sequence-211566-       10344 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 10344)
  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 10344)
  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: LT3GECIR
FEATURES             Location/Qualifiers
     source          1..10344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    113..346
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             531..575
                     /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    842..959
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        975..1353
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    983..1001
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1019..1037
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1055..1073
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1091..1109
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1127..1145
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1163..1181
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1199..1217
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1262..1286
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(1377..1379,1380..1382,1383..2096)
                     /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(1422..1443)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1683..1702)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2030..2051
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           2290..2304
                     /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        2483..2982
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2518..2541)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2915..2934
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3005..3715
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3150..3168)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3418..3437
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3735..4302
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3752..4304
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3896..3913)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4123..4142
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4316..4933,4934..4939,4940..4975,4976..4978,4979..5014,
                     5015..5017,5018..5053,5054..5062)
                     /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    5108..5696
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5161..5181)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5579..5590)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5903..6136
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6208..6329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6245..6264)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6299..6318
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6369..6504
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6524..6543)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6525..6543)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6553..6570)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6553..6569)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6562..6584)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6711..7166
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6798..6817)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7008..7029
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7192..7296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7297..7365,7366..8157)
                     /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(7515..7534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8328..8916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8817..8836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9070..9087
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9204..9225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9240..9245,9246..9263,9264..9270)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9278..9294
                     /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     9283..9305
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9302..9318
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9302..9318
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9337..9357
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9339..9357
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9385..9611
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9612..9792
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9839..9964
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
       61 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
      121 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
      181 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
      241 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
      301 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
      361 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
      421 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
      481 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
      541 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
      601 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
      661 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
      721 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
      781 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcggttaa
      841 cttttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
      901 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta
      961 tcgattctag attcgagttt actccctatc agtgatagag aacgtatgaa gagtttactc
     1021 cctatcagtg atagagaacg tatgcagact ttactcccta tcagtgatag agaacgtata
     1081 aggagtttac tccctatcag tgatagagaa cgtatgacca gtttactccc tatcagtgat
     1141 agagaacgta tctacagttt actccctatc agtgatagag aacgtatatc cagtttactc
     1201 cctatcagtg atagagaacg tataagcttt aggcgtgtac ggtgggcgcc tataaaagca
     1261 gagctcgttt agtgaaccgt cagatcgcct ggagcaattc cacaacactt ttgtcttata
     1321 ccaactttcc gtaccacttc ctaccctcgt aaagtcgagc ttgcgttgga tccaccatgg
     1381 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1441 acgtaaacgg ccacaagttc agcgtgtctg gcgagggcga gggcgatgcc acctacggca
     1501 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1561 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1621 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1681 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1741 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1801 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1861 tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1921 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1981 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2041 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatgac
     2101 gtacggcggc cgcctcgact agggataaca gggtaattgt ttgaatgagg cttcagtact
     2161 ttacagaatc gttgcctgca catcttggaa acacttgctg ggattacttc gacttcttaa
     2221 cccaacagaa ggctcgagaa ggtatattgc tgttgacagt gagcgcagga attataatgc
     2281 ttatctatag tgaagccaca gatgtataga taagcattat aattcctatg cctactgcct
     2341 cggacttcaa ggggctagaa ttcgagcaat tatcttgttt actaaaactg aataccttgc
     2401 tatctctttg atacattttt acaaagctga attaaaatgg tataaattaa atcacttttt
     2461 tcaattgacg cgtaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
     2521 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
     2581 cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca
     2641 ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg
     2701 acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag
     2761 caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc
     2821 tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc
     2881 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     2941 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cccaagcttc
     3001 caccatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     3061 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     3121 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     3181 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     3241 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     3301 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     3361 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     3421 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     3481 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     3541 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     3601 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     3661 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtaagcggc
     3721 cgcatgcatc caattccgcc ccccccccct aacgttactg gccgaagccg cttggaataa
     3781 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     3841 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     3901 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     3961 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     4021 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     4081 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     4141 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     4201 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     4261 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatgag
     4321 tagactggac aagagcaaag tcataaacgg agctctggaa ttactcaatg gtgtcggtat
     4381 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta
     4441 ctggcacgtg aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag
     4501 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa
     4561 cgccaagtca taccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg
     4621 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca
     4681 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact
     4741 gggctgcgta ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac
     4801 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc ggcagggagc
     4861 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa
     4921 gtgcgaaagc ggcgggccga ccgacgccct tgacgatttt gacttagaca tgctcccagc
     4981 cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga
     5041 ccttgacatg ctccccgggt aatccggact cgacctgcag ccaagcttat cgaaacgcgt
     5101 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5161 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5221 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5281 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5341 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5401 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5461 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     5521 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     5581 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     5641 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     5701 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     5761 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     5821 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     5881 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     5941 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6001 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6061 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6121 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     6181 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     6241 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6301 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     6361 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6421 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6481 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     6541 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     6601 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     6661 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     6721 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6781 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6841 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6901 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6961 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     7021 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     7081 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     7141 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     7201 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7261 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7321 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7381 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7441 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7501 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7561 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7621 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7681 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7741 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7801 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7861 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7921 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7981 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     8041 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     8101 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8161 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8221 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8281 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8341 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8401 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8461 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8521 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8581 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8641 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8701 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8761 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8821 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8881 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8941 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     9001 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     9061 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     9121 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     9181 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     9241 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     9301 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     9361 caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca
     9421 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
     9481 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
     9541 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
     9601 gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     9661 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     9721 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     9781 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     9841 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     9901 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     9961 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
    10021 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
    10081 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
    10141 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
    10201 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
    10261 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
    10321 ctggaggagg agatatgagg gaca
//
