LOCUS       sequence-266074-       10296 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 10296)
  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 10296)
  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: LT3REVIR
FEATURES             Location/Qualifiers
     source          1..10296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1010..1030
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1058..1284
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1285..1465
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1512..1637
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2130..2363
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2548..2592
                     /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    2859..2976
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2994..3372
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    3002..3020
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3038..3056
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3074..3092
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3110..3128
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3146..3164
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3182..3200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3218..3236
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3281..3305
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3396..4073
                     /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(3576..3596)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3988..4011
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     ncRNA           4257..4271
                     /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        4450..4949
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4485..4508)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     4882..4901
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(4972..4974,4975..4977,4978..5691)
                     /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(5017..5038)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5278..5297)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5625..5646
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5704..6271
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5721..6273
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5865..5882)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6092..6111
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6285..6902,6903..6908,6909..6944,6945..6947,6948..6983,
                     6984..6986,6987..7022,7023..7031)
                     /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    7077..7665
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7130..7150)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7548..7559)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7872..8105
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8177..8298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8214..8233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8268..8287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8338..8473
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8493..8512)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8494..8512)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8522..8539)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8522..8538)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8531..8553)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8680..9135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8767..8786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8977..8998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9161..9265
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9266..9334,9335..10126)
                     /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(9484..9503)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     1081 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     1141 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     1201 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     1261 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     1321 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1381 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1441 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     1501 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     1561 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     1621 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     1681 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     1741 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     1801 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     1861 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     1921 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     1981 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2041 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     2101 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     2161 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     2221 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     2281 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     2341 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     2401 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     2461 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     2521 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     2581 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     2641 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     2701 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     2761 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     2821 attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt
     2881 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2941 gaattacaaa aacaaattac aaaaattcaa aattttatcg atcctctaga ttcgagttta
     3001 ctccctatca gtgatagaga acgtatgaag agtttactcc ctatcagtga tagagaacgt
     3061 atgcagactt tactccctat cagtgataga gaacgtataa ggagtttact ccctatcagt
     3121 gatagagaac gtatgaccag tttactccct atcagtgata gagaacgtat ctacagttta
     3181 ctccctatca gtgatagaga acgtatatcc agtttactcc ctatcagtga tagagaacgt
     3241 ataagcttta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta gtgaaccgtc
     3301 agatcgcctg gagcaattcc acaacacttt tgtcttatac caactttccg taccacttcc
     3361 taccctcgta aagtcgagct tgcgttggat ccaccatggc ctcctccgag aacgtcatca
     3421 ccgagttcat gcgcttcaag gtgcgcatgg agggcaccgt gaacggccac gagttcgaga
     3481 tcgagggcga gggcgagggc cgcccctacg agggccacaa caccgtgaag ctgaaggtga
     3541 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc cagtacggct
     3601 ccaaggtgta cgtgaagcac cccgccgaca tccccgacta caagaagctg tccttccccg
     3661 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggcg accgtgaccc
     3721 aggactcctc cctgcaggac ggctgcttca tctacaaggt gaagttcatc ggcgtgaact
     3781 tcccctccga cggccccgtg atgcagaaga agaccatggg ctgggaggcc tccaccgagc
     3841 gcctgtaccc ccgcgacggc gtgctgaagg gcgagaccca caaggccctg aagctgaagg
     3901 acggcggcca ctacctggtg gagttcaagt ccatctacat ggccaagaag cccgtgcagc
     3961 tgcccggcta ctactacgtg gacgccaagc tggacatcac ctcccacaac gaggactaca
     4021 ccatcgtgga gcagtacgag cgcaccgagg gccgccacca cctgttcctg tagcggccgc
     4081 ctcgactagg gataacaggg taattgtttg aatgaggctt cagtacttta cagaatcgtt
     4141 gcctgcacat cttggaaaca cttgctggga ttacttcgac ttcttaaccc aacagaaggc
     4201 tcgagaaggt atattgctgt tgacagtgag cgcaggaatt ataatgctta tctatagtga
     4261 agccacagat gtatagataa gcattataat tcctatgcct actgcctcgg acttcaaggg
     4321 gctagaattc gagcaattat cttgtttact aaaactgaat accttgctat ctctttgata
     4381 catttttaca aagctgaatt aaaatggtat aaattaaatc acttttttca attgacgcgt
     4441 aattctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag
     4501 ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca
     4561 ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc
     4621 ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg
     4681 aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg
     4741 taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc
     4801 tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc
     4861 gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt
     4921 tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttccac catggtgagc
     4981 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     5041 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     5101 accctgaagc tgatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     5161 accctgggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac
     5221 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     5281 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     5341 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     5401 tacaactaca acagccacaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     5461 gccaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc cgaccactac
     5521 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     5581 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     5641 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta agaattgttc
     5701 aattccgccc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     5761 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     5821 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     5881 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     5941 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     6001 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     6061 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     6121 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     6181 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     6241 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatgagt agactggaca
     6301 agagcaaagt cataaacgga gctctggaat tactcaatgg tgtcggtatc gaaggcctga
     6361 cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga
     6421 agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc
     6481 acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat
     6541 accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa
     6601 cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct
     6661 ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat
     6721 tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta
     6781 tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct
     6841 tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg
     6901 gcgggccgac cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg
     6961 acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac cttgacatgc
     7021 tccccgggta atccggactc gacctgcagc caagcttatc gaaacgcgtc tggaacaatc
     7081 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     7141 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     7201 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     7261 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     7321 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     7381 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     7441 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     7501 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     7561 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     7621 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg
     7681 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
     7741 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
     7801 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     7861 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     7921 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7981 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     8041 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     8101 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     8161 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     8221 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     8281 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     8341 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     8401 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     8461 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     8521 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     8581 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     8641 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     8701 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     8761 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     8821 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     8881 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     8941 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     9001 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     9061 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     9121 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     9181 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     9241 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     9301 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     9361 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     9421 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     9481 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     9541 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     9601 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     9661 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     9721 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     9781 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     9841 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     9901 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     9961 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
    10021 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
    10081 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
    10141 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
    10201 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
    10261 agcgtcagac cccgtagaaa agatcaaagg atcttc
//
