LOCUS       sequence-13250-e        9041 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral transfer plasmid for Tet-regulated shRNA expression with 
            a TREtight promoter. Contains DsRed2 and Venus fluorescent 
            reporters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9041)
  AUTHORS   Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe 
            SW
  TITLE     Toolkit for evaluating genes required for proliferation and survival
            using tetracycline-regulated RNAi.
  JOURNAL   Nat Biotechnol. 2011 Jan . 29(1):79-83.
  PUBMED    21131983
REFERENCE   2  (bases 1 to 9041)
  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"; journalName: "Nat 
            Biotechnol. 2011 Jan . 29(1):79-83."
COMMENT     Sequence Label: TtRMPVIR
FEATURES             Location/Qualifiers
     source          1..9041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     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        1600..1914
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    1608..1626
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1644..1662
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1679..1697
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1715..1733
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1751..1769
                     /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     1887..1911
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1953..2630
                     /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(2133..2153)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2545..2568
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     ncRNA           2671..2792
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2820..2834
                     /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"
     ncRNA           2862..2988
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        3011..3510
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3046..3069)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3443..3462
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             join(3533..3535,3536..3538,3539..4252)
                     /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(3578..3599)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3839..3858)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4186..4207
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4265..4832
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4282..4834
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4426..4443)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4653..4672
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4846..5463,5464..5469,5470..5505,5506..5508,5509..5544,
                     5545..5547,5548..5583,5584..5592)
                     /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             5747..6172
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(6275..6297)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6433..6453)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6438..6767
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6618..6753
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6789..6808
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6889..6906)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7060..7648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7140..7159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7819..8610,8611..8679))
                     /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     8442..8461
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8680..8784)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8852..8870
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8970..9041,1..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 gataccgtcg agtttactcc ctatcagtga
     1621 tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgatgtc gagtttactc
     1681 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
     1741 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttatccct atcagtgata
     1801 gagaacgtat gtcgagttta ctccctatca gtgatagaga acgtatgtcg aggtaggcgt
     1861 gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
     1921 attaattcga gctcggtacc cggggatcca ccatggcctc ctccgagaac gtcatcaccg
     1981 agttcatgcg cttcaaggtg cgcatggagg gcaccgtgaa cggccacgag ttcgagatcg
     2041 agggcgaggg cgagggccgc ccctacgagg gccacaacac cgtgaagctg aaggtgacca
     2101 agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttccag tacggctcca
     2161 aggtgtacgt gaagcacccc gccgacatcc ccgactacaa gaagctgtcc ttccccgagg
     2221 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggcgacc gtgacccagg
     2281 actcctccct gcaggacggc tgcttcatct acaaggtgaa gttcatcggc gtgaacttcc
     2341 cctccgacgg ccccgtgatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc
     2401 tgtacccccg cgacggcgtg ctgaagggcg agacccacaa ggccctgaag ctgaaggacg
     2461 gcggccacta cctggtggag ttcaagtcca tctacatggc caagaagccc gtgcagctgc
     2521 ccggctacta ctacgtggac gccaagctgg acatcacctc ccacaacgag gactacacca
     2581 tcgtggagca gtacgagcgc accgagggcc gccaccacct gttcctgtag cggccagcta
     2641 agcctcgact agggataaca gggtaattgt ttgaatgagg cttcagtact ttacagaatc
     2701 gttgcctgca catcttggaa acacttgctg ggattacttc ttcaggttaa cccaacagaa
     2761 ggctcgagaa ggtatattgc tgttgacagt gagcgcagga attataatgc ttatctatag
     2821 tgaagccaca gatgtataga taagcattat aattcctatg cctactgcct cggaattcaa
     2881 ggggctactt taggagcaat tatcttgttt actaaaactg aataccttgc tatctctttg
     2941 atacattttt acaaagctga attaaaatgg tataaattaa atcacttttt tcaattctag
     3001 caattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca
     3061 gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc
     3121 accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct
     3181 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg
     3241 gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg
     3301 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg
     3361 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc
     3421 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg
     3481 ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttcca ccatggtgag
     3541 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3601 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3661 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3721 caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     3781 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3841 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3901 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     3961 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     4021 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     4081 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     4141 ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     4201 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagaattgtt
     4261 caattccgcc ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg
     4321 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     4381 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     4441 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     4501 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     4561 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     4621 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     4681 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca
     4741 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     4801 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatgag tagactggac
     4861 aagagcaaag tcataaacgg agctctggaa ttactcaatg gtgtcggtat cgaaggcctg
     4921 acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta ctggcacgtg
     4981 aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag gcatcatacc
     5041 cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa cgccaagtca
     5101 taccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg cacccgccca
     5161 acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca gcaaggcttc
     5221 tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact gggctgcgta
     5281 ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac caccgattct
     5341 atgcccccac ttctgagaca agcaattgag ctgttcgacc ggcagggagc cgaacctgcc
     5401 ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa gtgcgaaagc
     5461 ggcgggccga ccgacgccct tgacgatttt gacttagaca tgctcccagc cgatgccctt
     5521 gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga ccttgacatg
     5581 ctccccgggt aatccggact cgacctgcag ccaagcttat cgataccgtc gagatatcag
     5641 tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc
     5701 catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     5761 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     5821 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacaggg tcgaccctag
     5881 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     5941 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     6001 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     6061 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     6121 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
     6181 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggtaagctg
     6241 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     6301 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6361 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     6421 agatccggct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     6481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6541 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     6601 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6661 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6721 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttactggct
     6781 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     6841 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     6901 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7501 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7741 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7801 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7861 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7921 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7981 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8041 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8101 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
     8161 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8221 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8281 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8341 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8401 tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag
     8461 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8521 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8581 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8641 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8701 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8761 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8821 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8881 tcaagaatta gcttggccat tgcatacgtt gtatccatat cataatatgt acatttatat
     8941 tggctcatgt ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta
     9001 atcaattacg gggtcattag ttcatagccc atatatggag t
//
