LOCUS       sequence-209064-        8977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-regulated miR-E (miR-30 variant)-based RNAi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8977)
  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 8977)
  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: RT3REVIR
FEATURES             Location/Qualifiers
     source          1..8977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     464..484
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             514..689
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    752..951
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1152..1568
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1498..1520
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1536..1552
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        1605..1983
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1613..1631
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1649..1667
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1685..1703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1721..1739
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1757..1775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1793..1811
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1829..1847
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1892..1916
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2007..2684
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2187..2207)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2599..2622
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        2948..3447
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2983..3006)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3380..3399
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(3470..3472,3473..3475,3476..4189)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3515..3536)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3776..3795)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4123..4144
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4202..4768
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4219..4770
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4363..4380)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4590..4609
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4782..5399,5400..5405,5406..5441,5442..5444,5445..5480,
                     5481..5483,5484..5519,5520..5528)
                     /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"
     LTR             5683..6108
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6211..6233)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6369..6389)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6374..6703
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6554..6689
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6725..6744
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6825..6842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6996..7584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7076..7095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7755..8546,8547..8615))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8378..8397
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8616..8720)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8788..8806
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8906..8977,1..309)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgctctg gcattatgcc
      241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      481 cgtgtacggt gggaggtcta tataagcaga gctcaataaa agagcccaca acccctcact
      541 cggcgcgcca gtcttccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
      601 cttgctgttt gcatccgaat cgtggtctcg ctgttccttg ggagggtctc ctctgagtga
      661 ttgactaccc acgacggggg tctttcattt gggggctcgt ccgggatttg gagacccctg
      721 cccagggacc accgacccac caccgggagg taagctggcc agcaacttat ctgtgtctgt
      781 ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg tctgtactag ttagctaact
      841 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      901 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      961 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
     1021 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
     1081 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     1141 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1201 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1261 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1321 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1381 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1441 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1501 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1561 taggcgccgg aattgaagat ctgggggatc gatcctctag attcgagttt actccctatc
     1621 agtgatagag aacgtatgaa gagtttactc cctatcagtg atagagaacg tatgcagact
     1681 ttactcccta tcagtgatag agaacgtata aggagtttac tccctatcag tgatagagaa
     1741 cgtatgacca gtttactccc tatcagtgat agagaacgta tctacagttt actccctatc
     1801 agtgatagag aacgtatatc cagtttactc cctatcagtg atagagaacg tataagcttt
     1861 aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt cagatcgcct
     1921 ggagcaattc cacaacactt ttgtcttata ccaactttcc gtaccacttc ctaccctcgt
     1981 aaagtcgagc ttgcgttgga tccaccatgg cctcctccga gaacgtcatc accgagttca
     2041 tgcgcttcaa ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag atcgagggcg
     2101 agggcgaggg ccgcccctac gagggccaca acaccgtgaa gctgaaggtg accaagggcg
     2161 gccccctgcc cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt
     2221 acgtgaagca ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca
     2281 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggc gaccgtgacc caggactcct
     2341 ccctgcagga cggctgcttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg
     2401 acggccccgt gatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc
     2461 cccgcgacgg cgtgctgaag ggcgagaccc acaaggccct gaagctgaag gacggcggcc
     2521 actacctggt ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct
     2581 actactacgt ggacgccaag ctggacatca cctcccacaa cgaggactac accatcgtgg
     2641 agcagtacga gcgcaccgag ggccgccacc acctgttcct gtagcggccg cctcgactag
     2701 ggataacagg gtaattgttt gaatgaggct tcagtacttt acagaatcgt tgcctgcaca
     2761 tcttggaaac acttgctggg attacttcga cttcttaacc caacagaagg ctcgagcaac
     2821 cagaattcga gcaattatct tgtttactaa aactgaatac cttgctatct ctttgataca
     2881 tttttacaaa gctgaattaa aatggtataa attaaatcac ttttttcaat tgacgcgtaa
     2941 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
     3001 ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc
     3061 ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc
     3121 ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa
     3181 gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta
     3241 ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg
     3301 ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga
     3361 aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc
     3421 tcctcttcct catctccggg cctttcgacc tgcagcccaa gcttccacca tggtgagcaa
     3481 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3541 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3601 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3661 cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     3721 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3781 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3841 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3901 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     3961 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     4021 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     4081 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     4141 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag aattgttcaa
     4201 ttccgccccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     4261 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     4321 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     4381 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     4441 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     4501 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     4561 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     4621 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     4681 gctttacatg tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc acggggacgt
     4741 ggttttcctt tgaaaaacac gatgataata tggccacaac catgagtaga ctggacaaga
     4801 gcaaagtcat aaacggagct ctggaattac tcaatggtgt cggtatcgaa ggcctgacga
     4861 caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg cacgtgaaga
     4921 acaagcgggc cctgctcgat gccctgccaa tcgagatgct ggacaggcat catacccact
     4981 tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc aagtcatacc
     5041 gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag
     5101 agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc
     5161 tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc tgcgtattgg
     5221 aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc gattctatgc
     5281 ccccacttct gagacaagca attgagctgt tcgaccggca gggagccgaa cctgccttcc
     5341 ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg
     5401 ggccgaccga cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg
     5461 actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc
     5521 ccgggtaatc cggactcgac ctgcagccaa gcttatcgat accgtcgaga tatcagtggt
     5581 ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata
     5641 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
     5701 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     5761 gagaatagag aagttcagat caaggtcagg aacagatgga acagggtcga ccctagagaa
     5821 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     5881 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     5941 gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc
     6001 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     6061 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc
     6121 cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt aagctggctg
     6181 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6241 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6301 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtagat
     6361 ccggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     6421 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     6481 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     6541 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6601 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6661 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctta ctggcttaac
     6721 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     6781 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     6841 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6901 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6961 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7021 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7081 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7141 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7201 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7261 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7321 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7381 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7441 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7501 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7561 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7621 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7681 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     7741 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     7801 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     7861 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     7921 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7981 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     8041 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     8101 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     8161 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     8221 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     8281 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     8341 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     8401 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     8461 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     8521 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     8581 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     8641 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     8701 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     8761 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
     8821 gaattagctt ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc
     8881 tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca
     8941 attacggggt cattagttca tagcccatat atggagt
//
