LOCUS       sequence-354685-       10592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet Repressor reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10592)
  AUTHORS   Allen TP, Roennfeldt AE, Reckdharajkumar M, Liu M, Quinn RJ, Russell
            DL, Peet DJ, Whitelaw ML, Bersten DC
  TITLE     dFLASH; dual FLuorescent transcription factor Activity Sensor for 
            Histone integrated live-cell reporting and high-content screening
  JOURNAL   bioRxiv 2023.11.21.568191
REFERENCE   2  (bases 1 to 10592)
  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: "bioRxiv 
            2023.11.21.568191"
COMMENT     Sequence Label: pLV-REPORT(EF1a)TetR
FEATURES             Location/Qualifiers
     source          1..10592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    159..180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(195..200,201..218,219..225)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    233..249
                     /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     238..260
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     257..273
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     257..273
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     292..312
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        294..312
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        340..566
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             567..747
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    794..919
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1412..1645
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1830..1874
                     /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    2136..2253
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2325..2349
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2459..3079
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2532..2551)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        3276..4462
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3507..4453
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4410..4430
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4541..4550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4562..4582
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4586..4606
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4610..4630
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4640..4642,4643..4645,4646..5356,5357..5362,
                     5363..5482)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(4685..4706)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4946..4965)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5293..5314
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5483..5542
                     /codon_start=1
                     /product="2A peptide from equine rhinitis A virus 
                     polyprotein"
                     /label=E2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="QCTNYALLKLAGDVESNPGP"
     CDS             5543..6568
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5543..6568
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             7801..8034
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8106..8227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8143..8162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8197..8216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8329..8348
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(8613..8631)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8699..8803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8804..8872,8873..9664)
                     /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     complement(9022..9041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9835..10423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10324..10343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgataaccct aattcgatag catatgcttc ccgttgggta acatatgcta ttgaattagg
       61 gttagtctgg atagtatata ctactacccg ggaagcatat gctacccgtt tagggttcac
      121 cggtgatgcc ggccacgatg cgtccggcgt agaggatcta atgtgagtta gctcactcat
      181 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      241 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      301 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
      361 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
      421 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
      481 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
      541 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
      601 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
      661 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
      721 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
      781 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
      841 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
      901 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
      961 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     1021 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     1081 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     1141 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     1201 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     1261 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     1321 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     1381 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     1441 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     1501 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     1561 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     1621 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     1681 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     1741 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     1801 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     1861 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     1921 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     1981 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     2041 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     2101 ccattcgatt agtgaacgga tctcgacggt atcggtttta aaagaaaagg ggggattggg
     2161 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     2221 ttacaaaaac aaattacaaa aattcaaaat tttatcgatt tcgaacctag ggtttaaacg
     2281 gcgcgccagc tttaggcgtg tacggtgggc gcctataaaa gcagagctcg tttagtgaac
     2341 cgtcagatcg cctggagcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
     2401 ttcctaccct cgtaaagtcg acaccggggc ccagatctat cgatcggccg gatccaccat
     2461 gtctagatta gataaaagta aagtgattaa cagcgcatta gagctgctta atgaggtcgg
     2521 aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc agcctacatt
     2581 gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg agatgttaga
     2641 taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt ttttacgtaa
     2701 taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa aagtacattt
     2761 aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag cctttttatg
     2821 ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgctgtgg ggcattttac
     2881 tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa gggaaacacc
     2941 tactactgat agtatgccgc cattattacg acaagctatc gaattatttg atcaccaagg
     3001 tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag aaaaacaact
     3061 taaatgtgaa agtgggtccg cgtacagcgg atcccgggaa ttcagatctt attaacgccc
     3121 gccccacgac ccgcagcgcc cgaccgaaag gagcgaccgt cgacctcgac ggatcgggag
     3181 atctcgaccg agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct
     3241 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
     3301 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     3361 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     3421 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     3481 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     3541 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
     3601 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
     3661 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
     3721 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
     3781 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
     3841 aagatcgatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     3901 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     3961 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     4021 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     4081 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     4141 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     4201 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgat cgagcttttg
     4261 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     4321 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     4381 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     4441 tcttccattt caggtgtcgt gaggaattag cttgaccaag cttggtaccg agctcggatc
     4501 cactagtaac ggccgccagt gtgctggaat tctcgaggtc gccaccatgg tgcacgtgga
     4561 tccaaaaaag aagagaaagg tagatccaaa aaagaagaga aaggtagatc caaaaaagaa
     4621 gagaaaggta cacgtgtcca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4681 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4741 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4801 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4861 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4921 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4981 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     5041 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     5101 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     5161 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     5221 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     5281 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     5341 ggacgagctg tacaagaagc ttagccatgg cttcccgccg gaggtggagg agcaggatga
     5401 tggcacgctg cccatgtctt gtgcccagga gagcgggatg gaccgtcacc ctgcagcctg
     5461 tgcttctgct aggatcaatg tgcagtgtac taattatgct ctcttgaaat tggctggaga
     5521 tgttgagagc aacccaggtc ccatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     5581 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     5641 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     5701 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     5761 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
     5821 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     5881 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     5941 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     6001 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     6061 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     6121 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     6181 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     6241 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     6301 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     6361 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     6421 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     6481 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     6541 cagcactcgt ccgagggcaa aggaataggc tagcctagtg ccatttgttc agtggttcgt
     6601 agggctttcc cccactgttt ggctttcagt tatatggatg atgtggtatt gggggccaag
     6661 tctgtacagc atcttgagtc cctttttacc gctgttacca attttctttt gtctttgggt
     6721 atacatttaa accctaacaa aacaaagaga tggggttact ctctaaattt tatgggttat
     6781 gtcattggat gttatgggtc cttgccacaa gaacacatca tacaaaaaat caaagaatgt
     6841 tttagaaaac ttcctattaa caggcctatt gattggaaag tatgtcaacg aattgtgggt
     6901 cttttgggtt ttgctgcccc ttttacacaa tgtggttatc ctgcgttgat gcctttgtat
     6961 gcatgtattc aatctaagca ggctttcact ttctcgccaa cttacaaggc ctttctgtgt
     7021 aaacaatacc tgaaccttta ccccgttgcc cggcaacggc caggtctgtg ccaagtgttt
     7081 gctgacgcaa cccccactgg ctggggcttg gtcatgggcc atcagcgcat gcgtggaacc
     7141 ttttcggctc ctctgccgat ccatactgcg gaactcctag ccgcttgttt tgctcgcagc
     7201 aggtctggag caaacattat cgggactgat aactctgttg tcctatcccg caaatataca
     7261 tcgtttccat ggctgctagg ctgtgctgcc aactggatcc tgcgcgggac gtcctttgtt
     7321 tacgtcccgt cggcgctgaa tcctgcggac gacccttctc ggggtcgctt gggactctct
     7381 cgtccccttc tccgtctgcc gttccgaccg accacggggc gcacctctct ttacgcggac
     7441 tccccgtctg tgccttctca tctgccggac cgtgtgcact tcgcttcacc tctgcacgtc
     7501 gcatggagac caccgtgaac gcccaccaaa tattgcccaa ggtcttacat aagaggactc
     7561 ttggactctc agcaatgtca acgaccgacc ttgaggcata cttcaaagac tgtttgttta
     7621 aagactggga ggagttgggg gaggagatta ggttaaaggt ctttgtacta ggaggctgta
     7681 ggcataaatt ggtctgcgca ccagcaccat ggcgcaatca ctagagcggg gtacctttaa
     7741 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     7801 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     7861 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     7921 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     7981 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     8041 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     8101 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     8161 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     8221 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac
     8281 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     8341 ggccgaggcc ggatcccttg agtggctttc atcctggagc agactttgca gtctgtggac
     8401 tgcaacacaa cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg
     8461 ggacatgtac ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga
     8521 ggggcctgtg tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata
     8581 aggccccctt gttaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     8641 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     8701 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     8761 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     8821 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     8881 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     8941 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     9001 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
     9061 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     9121 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     9181 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     9241 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     9301 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
     9361 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     9421 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     9481 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     9541 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     9601 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     9661 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     9721 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     9781 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     9841 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     9901 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     9961 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
    10021 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
    10081 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
    10141 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
    10201 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
    10261 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
    10321 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
    10381 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
    10441 ctttttacgg ttcctggcct tttgctggcc ttgaagctgt ccctgatggt cgtcatctac
    10501 ctgcctggac agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat
    10561 cataatgggg aaggccatcc agcctcgcgt cg
//
