LOCUS       sequence-371645-        9933 bp    DNA     circular SYN 14-APR-2025
DEFINITION  A plasmid for dual expression of CRY2-mCh-superPLDx30 (an engineered
            PLD mutant with 30 times higher activity than the wild-type) and 
            plasma membrane-tagged CIBN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9933)
  AUTHORS   Tei R, Bagde SR, Fromme JC, Baskin JM
  TITLE     Activity-based directed evolution of a membrane editor in mammalian 
            cells.
  JOURNAL   Nat Chem. 2023 May 22. doi: 10.1038/s41557-023-01214-0.
  PUBMED    37217787
REFERENCE   2  (bases 1 to 9933)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem. 
            2023 May 22. doi: 10.1038/s41557-023-01214-0."
COMMENT     Sequence Label: pCDNA-CRY2-mCh-superPLDx30-P2A-CIBN-CAAX
FEATURES             Location/Qualifiers
     source          1..9933
                     /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    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             995..2488
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWRKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     CDS             2522..3226
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2664..2682)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2932..2951
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4817..4873
                     /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             4880..5389
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     promoter        complement(5486..5504)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5487..5504)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5524..5541)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5530..5754
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5800..6228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5887..5906)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6097..6118
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6237..6257)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6242..6571
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6422..6557
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6484..6503
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6638..7432
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6692..6711)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7302..7321
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7606..7727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7643..7662)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7697..7716
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7776..7792)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7776..7792)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7789..7811)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7800..7816
                     /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(7824..7830,7831..7848,7849..7854))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7869..7890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8007..8024)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8178..8766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8258..8277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8937..9728,9729..9797))
                     /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     9560..9579
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9798..9902)
                     /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 gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacgc caccatgaag atggacaaaa agactatagt
     1021 ttggtttaga agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga
     1081 aggatctgtt tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg
     1141 aagagcttca agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc
     1201 tcttggatct gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg
     1261 tatccgcgtt accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt
     1321 agttcgggac cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta
     1381 caatggagat ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac
     1441 gagtttcaat tcttactgga ggaaatgctt agatatgtcg attgaatccg ttatgcttcc
     1501 tcctccttgg cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat
     1561 tgaagaacta gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc
     1621 ttggtctcca ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt
     1681 gatagattat gcaaagaaca gcaagaaagt tgttgggaac tctacttcac tactttctcc
     1741 gtatctccat ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca
     1801 aattatatgg gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag
     1861 gggaatcggt ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga
     1921 gcaatcgttg ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa
     1981 ggcctggaga caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg
     2041 ggctaccgga tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt
     2101 tcttctcctt ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga
     2161 tttggaatgt gacatccttg gctggcagta tatctctggg agtatccccg atggccacga
     2221 gcttgatcgc ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta
     2281 cataaggcaa tggcttcccg agcttgcgag attgccaact gaatggattc atcatccatg
     2341 ggacgctcct ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa
     2401 acccattgta gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg
     2461 tgaagcacag atcatgatcg gagcagcagg tggaggcggt tcaggcggag gtggcagcgg
     2521 cgtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     2581 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     2641 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     2701 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     2761 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     2821 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     2881 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     2941 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3001 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     3061 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     3121 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     3181 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaaggctc gaggcgcagc
     3241 agcgggagca ggaggagcag gtcgatcggg atccggcgga tctgcagatt cagctacacc
     3301 acatttggac gccgttgagc aaacattgag gcaagtgtcc ccaggcctgg agggcgatgt
     3361 atgggaaaga actagtggaa ataaattgga cggaagtgcc gctgatccca gtgattggtt
     3421 gttgcaaacc ccaggatgtt ggggcgatga taaatgcgct gacagagttg ggacaaaaag
     3481 gttgctggct aagatgactg aaaatatagg gaatgccacc agaaccgtag atattagtac
     3541 acttgcacct ttcccaaacg gcgcattcca agacgctatt gtcgctggtt tgaaggagtc
     3601 cgcagctaga ggcaataagt tgaaagttag gatactggtt ggggctgcac ccgtctacca
     3661 catgaatgta ataccttcca agtataggga cgaattgact gcaaaacttg gcaaagccgc
     3721 agaaaacata actttgaacg tggctagtat gactacatca aaaacagctt tctcctggaa
     3781 ccatagcaaa atcttggttg tagacgggca aagcgccctt actggtggaa taaatagctg
     3841 gaaggatgat tacttggata ctactcatcc agtgtctgat gttgatcttg cactgaccgg
     3901 accagctgca ggctctgccg gtagatacct tgacactttg tggacatgga cctgccaaaa
     3961 caagagtaac atcgcttctg tctggtttgc agcttccggc aacgcagggt gcatggccac
     4021 tatgcataaa gatacaaacc ccaaggcatc acctgccacc gggaatgtcc cagtaatagc
     4081 cgttggaggg ttgggcgttg gcatcaaaga tgtggacccc aagtccactt tcaggcccga
     4141 tctgccaaca gctagcgata ctaagtgtgt ggtaggactt cacgacaata ctaacgccga
     4201 tagagactat gatacagtca acccagaaga gagtgccttg agagctcttg ttgcttcagc
     4261 aaaaagtcac attgaaatat ctcagcagga tttgaacgct acatgtccac ccttgcctag
     4321 gtatgacatt agattgtacg atgccttggc tgctaaaatg gctgctgggg ttaaggtgag
     4381 aattgtggtt agcgacccag ccaacagggg cgcagtgggc agtgtgggtt acagccaaat
     4441 aaaatcactt agtgaaatat cagacacttt gagaaacagg cttgcaaata tcacaggcgg
     4501 tcaacaagct gccaaaactg caatgtgtag caatttgcag cttgcaacct ttagatcctc
     4561 ccccaacgac aaatgggcag acgggcatcc ttatgcccaa caccataagt tggtttccgt
     4621 cgactcatcc acattttata tcggttcaaa aaatttgtac ccatcctggc tgcaagattt
     4681 tggctacata gtggaatccc cagaggctgc caagcaactg gatgccaaat tgcttgaccc
     4741 tcagtggaag tactcacagg agactgcaac agtggattac gcaagaggaa tctgcaacgc
     4801 tgaattcgga agcggcgcta caaactttag tctgctcaag caggctggcg atgtcgagga
     4861 aaacccaggt cccggtacca tgaatggagc tataggaggt gaccttttgc tcaattttcc
     4921 tgacatgtcg gtcctagagc gccaaagggc tcacctcaag tacctcaatc ccacctttga
     4981 ttctcctctc gccggcttct ttgccgattc ttcaatgatt accggcggcg agatggacag
     5041 ctatctttcg actgccggtt tgaatcttcc gatgatgtac ggtgagacga cggtggaagg
     5101 tgattcaaga ctctcaattt cgccggaaac gacgcttggg actggaaatt tcaaggcagc
     5161 gaagtttgat acagagacta aggattgtaa tgaggcggcg aagaagatga cgatgaacag
     5221 agatgaccta gtagaagaag gagaagaaga gaagtcgaaa ataacagagc aaaacaatgg
     5281 gagcacaaaa agcatcaaga agatgaaaca caaagccaag aaagaagaga acaatttctc
     5341 taatgattca tctaaagtga cgaaggaatt ggagaaaacg gattatattc atgtaccggt
     5401 cgccaccgga tcaggatcac tgtataaggg taaaaagaag aaaaagaagt caaagacaaa
     5461 gtgtgtaatt atgtaggggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     5521 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     5581 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5641 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5701 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     5761 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     5821 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5881 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5941 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     6001 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     6061 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     6121 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     6181 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     6241 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6301 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     6361 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     6421 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     6481 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     6541 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     6601 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     6661 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     6721 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     6781 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     6841 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     6901 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     6961 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     7021 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     7081 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     7141 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     7201 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     7261 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     7321 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     7381 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     7441 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     7501 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     7561 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     7621 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     7681 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     7741 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     7801 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     7861 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7921 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7981 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     8041 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     8101 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     8161 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     8221 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     8281 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     8341 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     8401 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     8461 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     8521 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     8581 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     8641 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     8701 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     8761 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     8821 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     8881 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     8941 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     9001 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     9061 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     9121 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     9181 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     9241 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     9301 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     9361 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     9421 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     9481 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     9541 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     9601 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     9661 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     9721 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     9781 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     9841 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     9901 cgcacatttc cccgaaaagt gccacctgac gtc
//
