LOCUS       sequence-346284-        8490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression and purification, fluorescently tagged scaffold
            for droplet visualization and volume quantification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8490)
  AUTHORS   Peeples W, Rosen MK
  TITLE     Mechanistic dissection of increased enzymatic rate in a 
            phase-separated compartment.
  JOURNAL   Nat Chem Biol. 2021 Jun;17(6):693-702. doi: 
            10.1038/s41589-021-00801-x. Epub 2021 May 25.
  PUBMED    34035521
REFERENCE   2  (bases 1 to 8490)
  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"; doi: 
            "10.1038/s41589-021-00801-x"; journalName: "Nat Chem Biol"; date: 
            "2021-06"; volume: "17"; issue: "6"; pages: "693-702"
COMMENT     Sequence Label: FRB-polySH33-EGFP
FEATURES             Location/Qualifiers
     source          1..8490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(277..296)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     437..459
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(453..644)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        1140..1217
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1218..2300
                     /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(1237..1256)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2543..2548,2549..2564,2565..2571)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2579..2595
                     /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     2584..2606
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             2665..3765
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      3622..3631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3739..3762
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             3814..3825
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3838..3858
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3871..4140
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     CDS             join(4924..4926,4927..5637)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4966..4987)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5227..5246)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5574..5595
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5644..5664
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5665..5682
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5695..5712)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5695..5711)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5704..5726)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5890..5907)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      6060..6146
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6238..6265
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        6284..6375
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6376..6444,6445..7236)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6594..6613)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7278..7791)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7473..7494)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7685..7704
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      7902..8490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8391..8410
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatgtgg
      241 tgcactctca gtacaccgca tgtggtgcac tctcagtaca atctgctctg atgccgcata
      301 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
      361 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
      421 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
      481 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gcagcgattc acagatgtct
      541 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
      601 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
      661 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
      721 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
      781 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
      841 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
      901 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
      961 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     1021 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     1081 aacgacagga gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgaaattccg
     1141 acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     1201 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     1261 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     1321 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     1381 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     1441 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     1501 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     1561 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     1621 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     1681 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     1741 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     1801 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     1861 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     1921 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     1981 gcgccattac cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg
     2041 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     2101 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     2161 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     2221 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2281 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
     2341 caattctcat gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca
     2401 ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
     2461 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
     2521 gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat gtgtggaatt
     2581 gtgagcggat aacaatttca cacaggaaac agccagtccg tttaggtgtt ttcacgagca
     2641 cttcaccaac aaggaccata gattatgaaa actgaagaag gtaaactggt aatctggatt
     2701 aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga gaaagatacc
     2761 ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc acaggttgcg
     2821 gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg tggctacgct
     2881 caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa gctgtatccg
     2941 tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat cgctgttgaa
     3001 gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac ctgggaagag
     3061 atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg
     3121 caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc gttcaagtat
     3181 gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc gaaagcgggt
     3241 ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac cgattactcc
     3301 atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg cccgtgggca
     3361 tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc gaccttcaag
     3421 ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg
     3481 aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga aggtctggaa
     3541 gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga ggaagagttg
     3601 gcgaaagatc cacgtattgc cgccaccatg gaaaacgccc agaaaggtga aatcatgccg
     3661 aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat caacgccgcc
     3721 agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaattc gagctcgaac
     3781 aacaacaaca ataacaataa caacaacctc gggatcgagg gaaggatttc agaatttgaa
     3841 aacctgtatt ttcagggcga attcatgtta gagatgtggc atgaaggact tgaggaagct
     3901 tcccgccttt actttggtga acgtaatgtt aagggcatgt tcgaagtatt ggaacccctg
     3961 cacgcgatga tggaacgtgg gccacagaca ttgaaagaaa caagctttaa ccaagcttac
     4021 ggacgtgatc tgatggaggc gcaagagtgg tgccgtaagt atatgaagtc cggtaacgta
     4081 aaagacctga cacaagcatg ggatttgtac taccatgttt ttcgccgtat ctcgaagcaa
     4141 gttgatggag gtagtggcgg atcaggtggg agtggagggt ctcatatgga cctcaacatg
     4201 cccgcttatg tgaaatttaa ctacatggct gagagagagg atgaattatc attgataaag
     4261 gggacaaagg tgatcgtcat ggagaaaagc agtgatgggt ggtggcgtgg tagctacaat
     4321 ggacaagttg gatggttccc ttcaaactat gtaactgaag aaggtgacag tcctttggcg
     4381 tctggtgcgg gtggttctga aggtggtggt tctgaaggtg gtacctctgg tgcgaccgac
     4441 ctcaacatgc ccgcttatgt gaaatttaac tacatggctg agagagagga tgaattatca
     4501 ttgataaagg ggacaaaggt gatcgtcatg gagaaaagca gtgatgggtg gtggcgtggt
     4561 agctacaatg gacaagttgg atggttccct tcaaactatg taactgaaga aggtgacagt
     4621 cctttggcat ccggagcagg aggctccgaa ggaggaggct ccgagggagg aacatccgga
     4681 gcaacagacc tcaacatgcc cgcttatgtg aaatttaact acatggctga gagagaggat
     4741 gaattatcat tgataaaggg gacaaaggtg atcgtcatgg agaaaagcag tgatgggtgg
     4801 tggcgtggta gctacaatgg acaagttgga tggttccctt caaactatgt aactgaagaa
     4861 ggtgacagtc ctttgggagg cggatctgga ggtagtggcg gatcaggtgg gagtggaggg
     4921 tctgtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4981 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     5041 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     5101 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     5161 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     5221 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     5281 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     5341 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     5401 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     5461 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     5521 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     5581 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaaggga
     5641 tccgaaaacc tgtattttca gggccatcat catcatcatc actgaaagct tggcactggc
     5701 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5761 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5821 ccaacagttg cgcagcctga atggcgaatg gcagcttggc tgttttggcg gatgagataa
     5881 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     5941 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     6001 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     6061 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     6121 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     6181 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     6241 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa
     6301 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6361 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6421 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6481 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6541 gagagttttc gccccgaaga acgtttccca atgatgagca cttttaaagt tctgctatgt
     6601 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     6661 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6721 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6781 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6841 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6901 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6961 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7021 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7081 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7141 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7201 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7261 atactttaga ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg
     7321 tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     7381 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     7441 aagaatagcc cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     7501 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     7561 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     7621 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     7681 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     7741 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc
     7801 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7861 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7921 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7981 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     8041 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     8101 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     8161 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     8221 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     8281 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     8341 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     8401 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     8461 tgctcgtcag gggggcggag cctatggaaa
//
