LOCUS       sequence-8480-ex       10227 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 10227)
  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 10227)
  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 Cre
FEATURES             Location/Qualifiers
     source          1..10227
                     /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"
     enhancer        462..765
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        766..969
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             1022..2101
                     /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(1041..1060)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             2117..2137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          2299..2364
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    2523..2644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2677..2693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2701..2717
                     /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(2725..2731,2732..2749,2750..2755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2770..2791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     3026..3042
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3049..3067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3157..3175
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     primer_bind     complement(3187..3206)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(3357..3377)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        complement(3523..3541)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3524..3541)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3559..3575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3583..3599
                     /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(3607..3613,3614..3631,3632..3637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3652..3673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3790..3807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4015..4031
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4098..4401
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4402..4605
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    4723..4742
                     /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    4766..4785
                     /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    4809..4828
                     /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."
     primer_bind     5272..5291
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        5319..5337
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5348..5968
                     /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(5421..5440)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     polyA_signal    6010..6131
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(6253..7284)
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     protein_bind    7642..7661
                     /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    7685..7704
                     /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    7728..7747
                     /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(7821..8082)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     rep_origin      complement(8408..8996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8488..8507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9167..9958,9959..10027))
                     /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     9790..9809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10028..10132)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10200..10218
                     /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 ctagttatta atagtaatca
      421 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      481 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      541 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      601 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      661 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      721 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      781 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      841 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      901 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      961 agcagagctg gtttagtgaa ccgtcagatc ggatcctcta gagtcgacca ttgccgccac
     1021 catgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt
     1081 ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc
     1141 ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca
     1201 gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt
     1261 cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga
     1321 acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag
     1381 tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg
     1441 cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat
     1501 tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca
     1561 gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg
     1621 ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg
     1681 gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac
     1741 tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc
     1801 cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc
     1861 atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag
     1921 cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc
     1981 cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg
     2041 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     2101 gagcagcctg aggcctccta agaagaagag gaaggtttga gcgcaacgca attaatgtaa
     2161 gttagctcac tcattaggca ccccaggctt tacactttat gcttccgacc tgcagcccct
     2221 tggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca
     2281 gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg
     2341 attctaattg tttgtgtatt ttagatcaca gtcccaaggc tcatttcagg cccctcagtc
     2401 ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     2461 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     2521 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     2581 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     2641 cttatcatgt ctgggcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2701 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2761 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2821 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2881 tttgcgtatt gggcgctctt ccgctgcaac tgttgggaag ggcgatcggt gcgggcctct
     2941 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     3001 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
     3061 ctatagggcg aattgggcct ttttatacaa aggatatcag gtgttcgcac ctgatatcct
     3121 ttgtatcggt gtttcgtcct ttccacaaga tatataactc tatcaatgat agagtacttt
     3181 caagttacgg taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac
     3241 ccaagaaatt attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc
     3301 aaattaattc taattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc
     3361 atgggaaata ggccctcttc ctgcccgacc ttggcgcgcg ctcggcgcgc ggtcacgctc
     3421 cgtcacgtgg tgcgttttgc ctaatcacta gtgaattcgc ggccgcctgc aggtcgacca
     3481 tatgggagag ctcccaacgc gttggatgca tagcttgagt attctatagt gtcacctaaa
     3541 tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3601 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3661 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3721 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3781 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3841 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3901 catggccggc ttaatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     3961 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     4021 cgacggccag tgaattctag ttattaatag taatcaatta cggggtcatt agttcatagc
     4081 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     4141 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     4201 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     4261 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     4321 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     4381 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     4441 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     4501 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     4561 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
     4621 cagatcggat cctctagagt cgacctgcag gatctaagct tggacaaact acctacagag
     4681 atttaaagct ctaaggtaaa tataaaattt actaggttgt ggaattgtga gcgctcacaa
     4741 ttccacagtc gaccctaggt tgtggaattg tgagcgctca caattccaca gtcgacccta
     4801 ggttgtggaa ttgtgagcgc tcacaattcc acagtcgacc ctagtgtata atgtgttaaa
     4861 ctacggatcc gtctcccatt aggcctacaa tggtgagaca agtagccaac agggaagggt
     4921 tgcaaatatc atttgggcac acctatgata atattgatga agcagacagt attcagcaag
     4981 taactgagag gtgggaagct caaagccaaa gtcctaatgt gcagtcaggt gaatttattg
     5041 aaaaatttga ggctcctggt ggtgcaaatc aaagaactgc tcctcaggga tcctaattgt
     5101 ttgtgtattt tagattccaa ccaagcttgc ggccgctcag gagctaagga agctaaaatg
     5161 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacgg
     5221 gcccctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     5281 tggttattgt gctgtctcat cattttggca aagaattgta atacgactca ctatagggcg
     5341 aattgatatg tctagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa
     5401 tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca
     5461 gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga
     5521 gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt
     5581 tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa
     5641 agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc
     5701 ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca gcgctgtggg
     5761 gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag
     5821 ggaaacacct actactgata gtatgccgcc attattacga caagctatcg aattatttga
     5881 tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga
     5941 aaaacaactt aaatgtgaaa gtgggtccgc gtacagcgga tcccgggaat tcagatctta
     6001 ttaaagcaga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6061 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6121 aatgtatctt atcatgtctg gtcgaatcca tcacactggc ggccgctcga ggcagtgaaa
     6181 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttacacctga
     6241 tatcctttgt atctaatcgc catcttccag caggcgcacc attgcccctg tttcactatc
     6301 caggttacgg atatagttca tgacaatatt tacattggtc cagccaccag cttgcatgat
     6361 ctccggtatt gaaactccag cgcgggccat atctcgcgcg gctccgacac gggcactgtg
     6421 tccagaccag gccaggtatc tctgaccaga gtcatcctta gcgccgtaaa tcaatcgatg
     6481 agttgcttca aaaatccctt ccagggcgcg agttgatagc tggctggtgg cagatggcgc
     6541 ggcaacacca ttttttctga cccggcaaaa caggtagtta ttcggatcat cagctacacc
     6601 agagacggaa atccatcgct cgaccagttt agttaccccc aggctaagtg ccttctctac
     6661 acctgcggtg ctaaccagcg ttttcgttct gccaatatgg attaacattc tcccaccgtc
     6721 agtacgtgag atatctttaa ccctgatcct ggcaatttcg gctatacgta acagggtgtt
     6781 ataagcaatc cccagaaatg ccagattacg tatatcctgg cagcgatcgc tattttccat
     6841 gagtgaacga acctggtcga aatcagtgcg ttcgaacgct agagcctgtt ttgcacgttc
     6901 accggcatca acgttttctt ttcggatccg ccgcataacc agtgaaacag cattgctgtc
     6961 acttggtcgt ggcagcccgg accgacgatg aagcatgttt agctggccca aatgttgctg
     7021 gatagttttt actgccagac cgcgcgcctg aagatataga agataatcgc gaacatcttc
     7081 aggttctgcg ggaaaccatt tccggttatt caacttgcac catgccgccc acgaccggca
     7141 aacggacaga agcattttcc aggtatgctc agaaaacgcc tggcgatccc tgaacatgtc
     7201 catcaggttc ttgcgaacct catcactcgt tgcatcgacc ggtaatgcag gcaaattttg
     7261 gtgtacggtc agtaaattgg acatttcaca ctcctgcagg tcgactctag aggatcaatt
     7321 ccttagctcc tgagcggccg caagcttggt tggaatctaa aatacacaaa caattaggat
     7381 ccctgaggag cagttctttg atttgcacca ccaggagcct caaatttttc aataaattca
     7441 cctgactgca cattaggact ttggctttga gcttcccacc tctcagttac ttgctgaata
     7501 ctgtctgctt catcaatatt atcataggtg tgcccaaatg atatttgcaa cccttccctg
     7561 ttggctactt gtctcaccat tgtaggccta atgggagacg gatccgtagt ttaacacatt
     7621 atacactagg gtcgactgtg gaattgtgag cgctcacaat tccacaacct agggtcgact
     7681 gtggaattgt gagcgctcac aattccacaa cctagggtcg actgtggaat tgtgagcgct
     7741 cacaattcca caacctagta aattttatat ttaccttaga gctttaaatc tctgtaggta
     7801 gtttgtccaa gcttagatct ggtgcacacc aatgtggtga atggtcaaat ggcgtttatt
     7861 gtatcgagct aggcacttaa atacaatatc tctgcaatgc ggaattcagt ggttcgtcca
     7921 atccatgtca gacccgtctg ttgccttcct aataaggcac gatcgtacca ccttacttcc
     7981 accaatcggc atgcacggtg ctttttctct ccttgtaagg catgttgcta actcatcgtt
     8041 accatgttgc aagactacaa gagtattgca taagactaca tttccccctc cctatgcaaa
     8101 agcgaaacta ctatatcctg aggggactcc taaccgcgta caaccgaagc cccgcttttc
     8161 gcctaaacac accctagtcc cctcagatac gcgtatatct ggcccgtaca tcgcgaagca
     8221 gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag ccttgccttg
     8281 ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa gcagggagca
     8341 gccatgtcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     8401 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     8461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     8521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     8581 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     8641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     8701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     8761 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8821 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     8881 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8941 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     9001 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     9061 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     9121 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     9181 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     9241 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     9301 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     9361 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     9421 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     9481 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     9541 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     9601 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     9661 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     9721 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9781 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9841 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9901 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9961 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    10021 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    10081 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    10141 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
    10201 ataggcgtat cacgaggccc tttcgtc
//
