LOCUS       sequence-8426-ex       11870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11870)
  AUTHORS   Deans TL, Cantor CR, Collins JJ
  TITLE     A tunable genetic switch based on RNAi and repressor proteins for 
            regulating gene expression in mammalian cells.
  JOURNAL   Cell. 2007 Jul 27. 130(2):363-72.
  PUBMED    17662949
REFERENCE   2  (bases 1 to 11870)
  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: "Cell. 2007 
            Jul 27. 130(2):363-72."
COMMENT     Sequence Label: LTRi EGFP
FEATURES             Location/Qualifiers
     source          1..11870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        663..924
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     protein_bind    998..1017
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    1041..1060
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    1084..1103
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     CDS             join(1459..1461,1462..1464,1465..2178)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1459..1461,1462..1464,1465..2175)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1504..1525)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1765..1784)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2112..2133
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2388..2404)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2412..2428
                     /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)."
     promoter        complement(join(2436..2442,2443..2460,2461..2466))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2481..2502
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     2737..2753
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2760..2778
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2873..2891
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     primer_bind     complement(2903..2922)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(3073..3093)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     enhancer        3235..3538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3539..3742
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             3795..4874
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3814..3833)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4890..4910
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          5072..5137
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    5296..5417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5450..5466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5474..5490
                     /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)."
     promoter        complement(join(5498..5504,5505..5522,5523..5528))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5543..5564
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5681..5698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5830..5863
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        5879..6378
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5914..5937)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6311..6330
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6392..7195
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6455..6474)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7065..7084
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7205..7668
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    7690..7723
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    7764..7885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7927..8547)
                     /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     8455..8474
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        complement(8558..8576)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8604..8623)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    9067..9086
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    9110..9129
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     protein_bind    9153..9172
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        complement(9290..9493)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        9494..9797
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(9864..9880)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(10051..10639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10131..10150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10810..11601,11602..11670))
                     /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     11433..11452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11671..11775)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     11843..11861
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt atcctgctcc ctgcttgtgt
      421 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
      481 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
      541 ggccagatat acgcgtatct gaggggacta gggtgtgttt aggcgaaaag cggggcttcg
      601 gttgtacgcg gttaggagtc ccctcaggat atagtagttt cgcttttgca tagggagggg
      661 gaaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
      721 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      781 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      841 ttccgcattg cagagatatt gtatttaagt gcctagctcg atacaataaa cgccatttga
      901 ccattcacca cattggtgtg caccagatct aagcttggac aaactaccta cagagattta
      961 aagctctaag gtaaatataa aatttactag gttgtggaat tgtgagcgct cacaattcca
     1021 cagtcgaccc taggttgtgg aattgtgagc gctcacaatt ccacagtcga ccctaggttg
     1081 tggaattgtg agcgctcaca attccacagt cgaccctagt gtataatgtg ttaaactacg
     1141 gatccgtctc ccattaggcc tacaatggtg agacaagtag ccaacaggga agggttgcaa
     1201 atatcatttg ggcacaccta tgataatatt gatgaagcag acagtattca gcaagtaact
     1261 gagaggtggg aagctcaaag ccaaagtcct aatgtgcagt caggtgaatt tattgaaaaa
     1321 tttgaggctc ctggtggtgc aaatcaaaga actgctcctc agggatccta attgtttgtg
     1381 tattttagat tccaaccaag cttgcggccg ctcaggagct aaggaattga tcctctagag
     1441 tcgacctgca gcacaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     1501 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
     1561 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     1621 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     1681 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     1741 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     1801 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     1861 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     1921 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     1981 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2041 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     2101 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2161 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
     2221 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     2281 tgaatgcact acacaaatca gcgattttaa tggttacaaa taaagcaata gcatcacaaa
     2341 tttcacaaat aaagcatttt tttcactgca agcttggcgt aatcatggtc atagctgttt
     2401 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     2461 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2521 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2581 gggagaggcg gtttgcgtat tgggcgctct tccgctgcaa ctgttgggaa gggcgatcgg
     2641 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     2701 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt
     2761 aatacgactc actatagggc gaattgggcc tttttctaca caaatcagcg attttctctt
     2821 gaaaaatcgc tgatttgtgt agcggtgttt cgtcctttcc acaagatata taactctatc
     2881 aatgatagag tactttcaag ttacggtaag catatgatag tccattttaa aacataattt
     2941 taaaactgca aactacccaa gaaattatta ctttctacgt cacgtatttt gtactaatat
     3001 ctttgtgttt acagtcaaat taattctaat tatctctcta acagccttgt atcgtatatg
     3061 caaatatgaa ggaatcatgg gaaataggcc ctcttcctgc ccgaccttgg cgcgcgctcg
     3121 gcgcgcggtc acgctccgtc acgtggtgcg ttttgcctaa tcactagtga attctagtta
     3181 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     3241 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3301 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3361 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3421 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     3481 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     3541 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     3601 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     3661 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     3721 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcggatcct ctagagtcga
     3781 ccattgccgc caccatgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     3841 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     3901 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     3961 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     4021 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     4081 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     4141 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     4201 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     4261 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     4321 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     4381 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     4441 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     4501 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     4561 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     4621 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
     4681 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     4741 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     4801 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     4861 tggaaagcgg gcagagcagc ctgaggcctc ctaagaagaa gaggaaggtt tgagcgcaac
     4921 gcaattaatg taagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     4981 acctgcagcc ccttggatct ttgtgaagga accttacttc tgtggtgtga cataattgga
     5041 caaactacct acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg
     5101 tgttaaacta ctgattctaa ttgtttgtgt attttagatc acagtcccaa ggctcatttc
     5161 aggcccctca gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag
     5221 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     5281 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5341 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5401 ctcatcaatg tatcttatca tgtctgggcc caagcttggc gtaatcatgg tcatagctgt
     5461 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5521 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5581 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5641 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5701 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5761 ccacagaatc aggggataac gcaggaaaga acatgacatg tggccctcta gactcgacac
     5821 gttttggccg aagttcctat tctctagaaa gtataggaac ttcgcggcca attctaccgg
     5881 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc
     5941 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc
     6001 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg
     6061 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt
     6121 ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg
     6181 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt
     6241 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc
     6301 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc
     6361 tcatctccgg gcctttcgac ctgcagccaa tatgggatcg gccattgaac aagatggatt
     6421 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     6481 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     6541 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     6601 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     6661 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     6721 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     6781 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     6841 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     6901 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     6961 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     7021 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     7081 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     7141 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagggga
     7201 tccgctgtaa gtctgcagaa attgatgatc tattaaacaa taaagatgtc cactaaaatg
     7261 gaagtttttc ctgtcatact ttgttaagaa gggtgagaac agagtaccta cattttgaat
     7321 ggaaggattg gagctacggg ggtgggggtg gggtgggatt agataaatgc ctgctcttta
     7381 ctgaaggctc tttactattg ctttatgata atgtttcata gttggatatc ataatttaaa
     7441 caagcaaaac caaattaagg gccagctcat tcctcccact catgatctat agatctatag
     7501 atctctcgtg ggatcattgt ttttctcttg attcccactt tgtggttcta agtactgtgn
     7561 ttnccaaatg tgtcagtttc atagcctgaa gaacgagatc agcagcctct gttccaacat
     7621 acacttcatt ctncagtatt gttttgccaa gttctaattc catcagaagc ttatcgatac
     7681 ccgtcgaggg aagttcctat tctctagaaa gtataggaac ttcgtcgagc ggccgccagt
     7741 gtgatggatt cgaccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     7801 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     7861 cattataagc tgcaataaac aagttctgct ttaataagat ctgaattccc gggatccgct
     7921 gtacgcggac ccactttcac atttaagttg tttttctaat ccgcatatga tcaattcaag
     7981 gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa
     8041 taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga cttgatgctc
     8101 ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg catataatgc
     8161 attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga gagtttcata
     8221 ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat gacttagtaa
     8281 agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt ccccttctaa
     8341 agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt cgagcaaagc
     8401 ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag
     8461 tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat
     8521 cactttactt ttatctaatc tagacatatc aattcgccct atagtgagtc gtattacaat
     8581 tctttgccaa aatgatgaga cagcacaata accagcacgt tgcccaggag ctgtaggaaa
     8641 aagaagaagg catgaacatg gttagcagag gggcccgttc tttacgatgc cattgggata
     8701 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgagc
     8761 ggccgcaagc ttggttggaa tctaaaatac acaaacaatt aggatccctg aggagcagtt
     8821 ctttgatttg caccaccagg agcctcaaat ttttcaataa attcacctga ctgcacatta
     8881 ggactttggc tttgagcttc ccacctctca gttacttgct gaatactgtc tgcttcatca
     8941 atattatcat aggtgtgccc aaatgatatt tgcaaccctt ccctgttggc tacttgtctc
     9001 accattgtag gcctaatggg agacggatcc gtagtttaac acattataca ctagggtcga
     9061 ctgtggaatt gtgagcgctc acaattccac aacctagggt cgactgtgga attgtgagcg
     9121 ctcacaattc cacaacctag ggtcgactgt ggaattgtga gcgctcacaa ttccacaacc
     9181 tagtaaattt tatatttacc ttagagcttt aaatctctgt aggtagtttg tccaagctta
     9241 gatcctgcag gtcgactcta gaggatccga tctgacggtt cactaaacca gctctgctta
     9301 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     9361 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     9421 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     9481 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     9541 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     9601 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     9661 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     9721 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     9781 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     9841 attactatta ataactagaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     9901 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     9961 agcgaagagg cccgcaccga ttaagccggc catgtgagca aaaggccagc aaaaggccag
    10021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
    10081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
    10141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
    10201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
    10261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
    10321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
    10381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
    10441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
    10501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
    10561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
    10621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
    10681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
    10741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
    10801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
    10861 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
    10921 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
    10981 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
    11041 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
    11101 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
    11161 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
    11221 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
    11281 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
    11341 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
    11401 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
    11461 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
    11521 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
    11581 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    11641 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    11701 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    11761 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
    11821 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
