LOCUS       sequence-209065-        9062 bp    DNA     circular SYN 14-APR-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 9062)
  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 9062)
  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"; 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: RT3GEPIR
FEATURES             Location/Qualifiers
     source          1..9062
                     /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        1598..1976
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1606..1624
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1642..1660
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1678..1696
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1714..1732
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1750..1768
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1786..1804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1822..1840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1885..1909
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(2000..2002,2003..2005,2006..2719)
                     /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(2045..2066)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2306..2325)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2653..2674
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           2913..2927
                     /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        3106..3605
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3141..3164)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3538..3557
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3626..4225
                     /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(3626..3645)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4122..4142
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4286..4853
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4303..4855
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4447..4464)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4674..4693
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4867..5484,5485..5490,5491..5526,5527..5529,5530..5565,
                     5566..5568,5569..5604,5605..5613)
                     /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             5768..6193
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6296..6318)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6454..6474)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6459..6788
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6639..6774
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6810..6829
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6910..6927)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7081..7669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7161..7180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7840..8631,8632..8700))
                     /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     8463..8482
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8701..8805)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8873..8891
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8991..9062,1..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 ctatcgattc tagattcgag tttactccct atcagtgata
     1621 gagaacgtat gaagagttta ctccctatca gtgatagaga acgtatgcag actttactcc
     1681 ctatcagtga tagagaacgt ataaggagtt tactccctat cagtgataga gaacgtatga
     1741 ccagtttact ccctatcagt gatagagaac gtatctacag tttactccct atcagtgata
     1801 gagaacgtat atccagttta ctccctatca gtgatagaga acgtataagc tttaggcgtg
     1861 tacggtgggc gcctataaaa gcagagctcg tttagtgaac cgtcagatcg cctggagcaa
     1921 ttccacaaca cttttgtctt ataccaactt tccgtaccac ttcctaccct cgtaaagtcg
     1981 agcttgcgtt ggatccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2041 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg
     2101 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2161 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2221 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2281 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2341 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2401 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2461 ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga
     2521 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2581 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2641 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2701 ggacgagctg tacaagtaat gacgtacggc ggccgcctcg actagggata acagggtaat
     2761 tgtttgaatg aggcttcagt actttacaga atcgttgcct gcacatcttg gaaacacttg
     2821 ctgggattac ttcgacttct taacccaaca gaaggctcga gaaggtatat tgctgttgac
     2881 agtgagcgca ggaattataa tgcttatcta tagtgaagcc acagatgtat agataagcat
     2941 tataattcct atgcctactg cctcggactt caaggggcta gaattcgagc aattatcttg
     3001 tttactaaaa ctgaatacct tgctatctct ttgatacatt tttacaaagc tgaattaaaa
     3061 tggtataaat taaatcactt ttttcaattg acgcgtaatt ctaccgggta ggggaggcgc
     3121 ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca
     3181 caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt
     3241 tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg
     3301 ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg
     3361 tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat
     3421 agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg
     3481 ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat
     3541 tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc
     3601 tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
     3661 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3721 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3781 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3841 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3901 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     3961 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4021 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4081 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     4141 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4201 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag
     4261 gagcgcacga ccccatgcat ccaattccgc cccccccccc taacgttact ggccgaagcc
     4321 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     4381 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     4441 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     4501 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     4561 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     4621 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     4681 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     4741 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     4801 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     4861 acaaccatga gtagactgga caagagcaaa gtcataaacg gagctctgga attactcaat
     4921 ggtgtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag
     4981 cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag
     5041 atgctggaca ggcatcatac ccacttctgc cccctggaag gcgagtcatg gcaagacttt
     5101 ctgcggaaca acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa
     5161 gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg
     5221 ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc
     5281 cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga
     5341 gagacaccta ccaccgattc tatgccccca cttctgagac aagcaattga gctgttcgac
     5401 cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag
     5461 aaacagctaa agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac
     5521 atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt
     5581 gacgattttg accttgacat gctccccggg taatccggac tcgacctgca gccaagctta
     5641 tcgataccgt cgagatatca gtggtccagg ctctagtttt gactcaacaa tatcaccagc
     5701 tgaagcctat agagtacgag ccatagataa aataaaagat tttatttagt ctccagaaaa
     5761 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt
     5821 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag
     5881 atggaacagg gtcgacccta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     5941 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     6001 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
     6061 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     6121 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     6181 ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac
     6241 ccaccaccgg gaggtaagct ggctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     6301 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6361 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     6421 cacgtagcga tagcggagtg tagatccggc tgtggaatgt gtgtcagtta gggtgtggaa
     6481 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6541 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6601 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6661 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6721 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6781 tttgcaaaaa gcttactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     6841 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
     6901 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6961 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7021 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7081 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7141 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7201 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7261 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7321 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7381 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7441 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7501 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7561 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7621 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7681 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7741 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7801 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7861 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7921 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7981 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8041 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8101 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg
     8161 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8221 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8281 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     8341 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     8401 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     8461 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8521 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8581 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8641 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8701 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8761 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8821 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     8881 tatcacgagg ccctttcgtc ttcaagaatt agcttggcca ttgcatacgt tgtatccata
     8941 tcataatatg tacatttata ttggctcatg tccaacatta ccgccatgtt gacattgatt
     9001 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     9061 gt
//
