LOCUS       sequence-334141-       10459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Equalizer-M plasmid that encodes eGFP to report circuit output 
            levels. This plasmid also encodes a separate mCherry expression 
            cassette to monitor plasmid dosage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10459)
  AUTHORS   Yang J, Lee J, Land MA, Lai S, Igoshin OA, St-Pierre F
  TITLE     A synthetic circuit for buffering gene dosage variation between 
            individual mammalian cells.
  JOURNAL   Nat Commun. 2021 Jul 5;12(1):4132. doi: 10.1038/s41467-021-23889-0.
  PUBMED    34226556
REFERENCE   2  (bases 1 to 10459)
  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 
            Commun."; date: "2021-07-5"; volume: "12(1):4132. doi"; pages: " 
            10.1038/s41467-021-23889-0"
COMMENT     Sequence Label: Equalizer-M plasmid with onboard mCherry cassette
FEATURES             Location/Qualifiers
     source          1..10459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    814..832
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    840..858
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     intron          1041..1613
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      1625..1634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2309..2365
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(2372..2374,2375..2377,2378..3091)
                     /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(2372..2374,2375..2377,2378..3088)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2417..2438)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2678..2697)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3025..3046
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           3297..3311
                     /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           3339..3467
                     /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"
     primer_bind     complement(3557..3577)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(3930..3947)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3936..4160
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4293..4312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4444..4491
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4499..5521
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4499..5521
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5651..5772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5688..5707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5742..5761
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5821..5837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5821..5837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5834..5856)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5845..5861
                     /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(5869..5875,5876..5893,5894..5899))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5914..5935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6052..6069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6182..7360
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          6413..7351
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     7308..7328
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      7399..7408
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7405..8115
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(7550..7568)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7818..7837
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(8207..8226)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    8272..8327
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(8326..8345)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(8704..9292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8784..8803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9463..10254,10255..10323))
                     /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     10086..10105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10324..10428)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcatccctat cagtgataga gatcagatct
      841 ccctatcagt gatagagagc tgttttgacc tccatagaag acaccgggac cgatccagcc
      901 tccggactct agcgtttaaa cttaagctgg gtacccgggg atcctctagg gcctctgagc
      961 tattccagaa gtagtgaaga ggcttttttg gaggcctagg cttttgcaaa aagctccgga
     1021 tcgatcctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta
     1081 ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa
     1141 gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct
     1201 gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact
     1261 ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt
     1321 actttctcta atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac
     1381 agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt
     1441 ctggctggcg tggaaatatt cttattggta gaaacaacta catcctggtc atcatcctgc
     1501 ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt
     1561 ccaaaccggg cccctctgct aaccatgttc atgccttctt ctttttccta caggtcctgc
     1621 aggcgccacc atggagtcta gactggacaa gagcaaagtc ataaactctg ctctggaatt
     1681 actcaatgaa gtcggtatcg aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt
     1741 tgagcagcct accctgtact ggcacgtgaa gaacaagcgg gccctgctcg atgccctggc
     1801 aatcgagatg ctggacaggc atcataccca cttctgcccc ctggaaggcg agtcatggca
     1861 agactttctg cggaacaacg ccaagtcatt ccgctgtgct ctcctctcac atcgcgacgg
     1921 ggctaaagtg catctcggca cccgcccaac agagaaacag tacgaaaccc tggaaaatca
     1981 gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac gcactgtacg ctctgtccgc
     2041 cgtgggccac tttacactgg gctgcgtatt ggaggatcag gagcatcaag tagcaaaaga
     2101 ggaaagagag acacctacca ccgattctat gcccccactt ctgagacaag caattgagct
     2161 gttcgaccat cagggagccg aacctgcctt ccttttcggc ctggaactaa tcatatgtgg
     2221 cctggagaaa cagctaaagt gcgaaagcgg cgggccaaaa aagaagagaa agggtgacgg
     2281 tgctggttta attaacatgg gaagcggagc tactaacttc agcctgctga agcaggctgg
     2341 agacgtggag gagaaccctg gacctgctag catggtgagc aagggcgagg agctgttcac
     2401 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     2461 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     2521 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     2581 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     2641 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     2701 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     2761 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     2821 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     2881 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2941 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     3001 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     3061 cactctcggc atggacgagc tgtacaagta gccgcttgaa gtctttaatt aaaaagcttg
     3121 tgagtatgtg ctcgcttcgg cagcacatat actatgtcga atgaggcttc agtactttac
     3181 agaatcgttg cctgcacatc ttggaaacac ttgctgggat tacttcttca ggttaaccca
     3241 acagaaggct cgagtgctgt tgacagtgag cgccgcttga agtctttaat taaatagtga
     3301 agccacagat gtatttaatt aaagacttca agcggtgcct actgcctcgg agaattcaag
     3361 gggctacttt aggagcaatt atcttgttta ctaaaactga ataccttgct atctctttga
     3421 tacattttta caaagctgaa ttaaaatggt ataaattaaa tcactttttt caattgtttc
     3481 cttttttttc ctcaggcggc cgcaatcaac ctctggatta caaaatttgt gaaagattga
     3541 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3601 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3661 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3721 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3781 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3841 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcctcgagtc
     3901 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3961 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4021 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4081 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4141 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     4201 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4261 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4321 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4381 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     4441 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     4501 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     4561 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     4621 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     4681 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     4741 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     4801 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     4861 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     4921 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     4981 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     5041 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     5101 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     5161 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     5221 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     5281 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     5341 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     5401 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     5461 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     5521 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5581 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5641 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5701 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5761 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5821 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5881 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5941 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6001 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6061 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6121 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtaag gagtgggaat
     6181 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     6241 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     6301 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     6361 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     6421 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     6481 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
     6541 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
     6601 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     6661 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     6721 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
     6781 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     6841 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     6901 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     6961 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     7021 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     7081 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     7141 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     7201 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     7261 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     7321 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggtaccggat cctctagagt
     7381 cgactccgga cgtcgacggc caccatggtg agcaagggcg aggaggataa catggccatc
     7441 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     7501 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     7561 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     7621 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     7681 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     7741 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     7801 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     7861 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     7921 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     7981 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     8041 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     8101 gagctgtaca agtaatgagc ggccgcttca ctcctcaggt gcaggctgcc tatcagaagg
     8161 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     8221 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     8281 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     8341 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     8401 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     8461 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     8521 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     8581 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     8641 tcacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     8701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     8761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     8821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     8881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     8941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     9001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     9061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     9121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     9181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     9241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     9301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     9361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     9661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     9721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     9781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     9841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     9901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     9961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    10021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    10081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    10141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    10201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    10261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    10321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    10381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    10441 aaaagtgcca cctgacgtc
//
