pC013 - Twinstrep-SUMO-huLwCas13a

LOCUS       pC013 - Twinstrep-SUMO-huLwCas13a                9542 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Twinstrep-SUMO-huLwCas13a for recombinant protein bacterial 
            expression. Insert is human codon optimized but expresses well in 
            bacteria..
ACCESSION   .
VERSION     .
KEYWORDS    pC013 - Twinstrep-SUMO-huLwCas13a
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9542)
  AUTHORS   Gootenberg JS, Abudayyeh OO, Lee JW, Essletzbichler P, Dy AJ, Joung 
            J, Verdine V, Donghia N, Daringer NM, Freije CA, Myhrvold C, 
            Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung DT, Sabeti PC, 
            Collins JJ, Zhang F
  TITLE     Nucleic acid detection with CRISPR-Cas13a/C2c2.
  JOURNAL   Science. 2017 Apr 13. pii: eaam9321. doi: 10.1126/science.aam9321.
  PUBMED    28408723
REFERENCE   2  (bases 1 to 9542)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9542
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(170..192)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    278..417
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     333..352
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(432..449)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(603..1191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(683..702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1362..2222)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1985..2004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2223..2327)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2395..2413
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2440..2468
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      2643..2690
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2686..2704
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2754..6209)
                     /codon_start=1
                     /product="Leptotrichia wadei CRISPR-Cas effector that acts 
                     as an RNA-guided RNAse (Abudayyeh et al., 2017)"
                     /label=LwCas13a
                     /note="previously called C2c2"
                     /translation="MKVTKVDGISHKKYIEEGKLVKSTSEENRTSERLSELLSIRLDIY
                     IKNPDNASEEENRIRRENLKKFFSNKVLHLKDSVLYLKNRKEKNAVQDKNYSEEDISEY
                     DLKNKNSFSVLKKILLNEDVNSEELEIFRKDVEAKLNKINSLKYSFEENKANYQKINEN
                     NVEKVGGKSKRNIIYDYYRESAKRNDYINNVQEAFDKLYKKEDIEKLFFLIENSKKHEK
                     YKIREYYHKIIGRKNDKENFAKIIYEEIQNVNNIKELIEKIPDMSELKKSQVFYKYYLD
                     KEELNDKNIKYAFCHFVEIEMSQLLKNYVYKRLSNISNDKIKRIFEYQNLKKLIENKLL
                     NKLDTYVRNCGKYNYYLQVGEIATSDFIARNRQNEAFLRNIIGVSSVAYFSLRNILETE
                     NENDITGRMRGKTVKNNKGEEKYVSGEVDKIYNENKQNEVKENLKMFYSYDFNMDNKNE
                     IEDFFANIDEAISSIRHGIVHFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLK
                     IFKQLNSANVFNYYEKDVIIKYLKNTKFNFVNKNIPFVPSFTKLYNKIEDLRNTLKFFW
                     SVPKDKEEKDAQIYLLKNIYYGEFLNKFVKNSKVFFKITNEVIKINKQRNQKTGHYKYQ
                     KFENIEKTVPVEYLAIIQSREMINNQDKEEKNTYIDFIQQIFLKGFIDYLNKNNLKYIE
                     SNNNNDNNDIFSKIKIKKDNKEKYDKILKNYEKHNRNKEIPHEINEFVREIKLGKILKY
                     TENLNMFYLILKLLNHKELTNLKGSLEKYQSANKEETFSDELELINLLNLDNNRVTEDF
                     ELEANEIGKFLDFNENKIKDRKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLLEK
                     IADKAKYKISLKELKEYSNKKNEIEKNYTMQQNLHRKYARPKKDEKFNDEDYKEYEKAI
                     GNIQKYTHLKNKVEFNELNLLQGLLLKILHRLVGYTSIWERDLRFRLKGEFPENHYIEE
                     IFNFDNSKNVKYKSGQIVEKYINFYKELYKDNVEKRSIYSDKKVKKLKQEKKDLYIRNY
                     IAHFNYIPHAEISLLEVLENLRKLLSYDRKLKNAIMKSIVDILKEYGFVATFKIGADKK
                     IEIQTLESEKIVHLKNLKKKKLMTDRNSEELCELVKVMFEYKALE"
     CDS             complement(6216..6506)
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
     CDS             complement(6507..6530)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(6567..6590)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(6603..6620)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(6630..6647)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             6666..6688
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    6703..6727
                     /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     complement(6727..6746)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6728..6746)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6830..6849)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7055..7132
                     /gene="lacI"
                     /label=lacI promoter
     CDS             7133..8215
                     /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(7152..7171)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             9527..176
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
       61 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
      121 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
      181 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
      241 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
      301 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
      361 gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
      421 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      481 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      541 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      601 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      661 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      721 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      781 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      841 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      901 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      961 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1021 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     1081 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1141 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1201 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1261 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1321 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1381 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1441 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1501 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1561 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1621 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca
     1681 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1741 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1801 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1861 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1921 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca
     1981 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     2041 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     2101 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     2161 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2221 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2281 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     2341 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     2401 cgtatcacga ggccctttcg tcttcaagaa ttctcatgtt tgacagctta tcatcgataa
     2461 gctttaatgc ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa
     2521 atctaacaat gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg
     2581 cttggttatg ccggtactgc cgggcctctt gcgggatatc cggatatagt tcctcctttc
     2641 agcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctag ttattgctca
     2701 gcggtggcag cagccaactc agcttccttt cgggcctcga gtgcggccgc ttattccagg
     2761 gccttgtact cgaacatgac tttcacgagt tcgcacagtt cctcgctgtt ccggtcggtc
     2821 atcagtttct ttttcttcag attcttcagg tgcacgatct tctctgattc cagggtctgg
     2881 atttcgatct tcttgtcagc gccgatcttg aaggtggcca cgaagccgta ttctttcaga
     2941 atgtccacga tggacttcat gatggcgttc ttcagcttcc ggtcgtagga cagcagcttc
     3001 cgcaggtttt ccagcacttc cagcaggcta atctcggcgt gggggatgta gttgaagtgg
     3061 gcaatgtagt tccggatgta caggtccttt ttttcctgct tcagtttctt cactttcttg
     3121 tcggagtaga tgctccgctt ttccacattg tccttgtaca gttctttgta gaagttgata
     3181 tacttttcca cgatctggcc gcttttgtac ttcacattct tggagttgtc gaaattgaaa
     3241 atttcctcga tgtagtggtt ctcgggaaac tcgcccttca gccggaatct caggtcccgc
     3301 tcccagatgc tggtgtagcc cacgagccgg tgcaggatct tcagcagcag gccctgcagc
     3361 aggttcagct cattgaattc caccttgttc ttcaggtggg tgtacttctg gatgttgccg
     3421 atggccttct catactcttt gtagtcctcg tcgttgaact tttcgtcctt cttgggtctg
     3481 gcgtacttcc ggtgcaggtt ctgctgcatg gtgtagttct tttcaatctc attcttcttg
     3541 ttgctgtact ctttcagttc tttcaggctg atcttatact tggccttatc ggcgatcttt
     3601 tccagcagat tcagcatgcc gtatttcttg atattgtaga aggcccggtg cttgatgatg
     3661 ttctcgccgt cgaaatagat cttgttggtg tcgaactttt tcagctcttt ccggtccttg
     3721 attttgtttt cgttgaagtc caggaacttg ccgatctcgt tggcttccag ctcgaagtcc
     3781 tcggtcactc tgttgttgtc caggttcagc aggttgatca gttccagctc gtcgctgaag
     3841 gtttcttctt tgttggcgga ctggtacttt tccaggctgc ccttcaggtt ggtcagctct
     3901 ttgtggttca gcagcttcag gatcaggtaa aacatgttca gattctcggt gtacttcaga
     3961 atcttcccca gcttgatctc gcgcacgaac tcattgatct cgtgagggat ttctttgttc
     4021 cgattgtgct tctcatagtt cttcaggatc ttgtcgtact tctctttgtt atcctttttg
     4081 atcttgatct tggagaagat gtcgttgttg tcattgttgt tgttgctctc gatatacttc
     4141 agattgttct tgttcaggta gtcgatgaag cccttcagga aaatctgctg aataaagtcg
     4201 atgtaggtat tcttttcctc tttgtcctgg ttgttgatca tctctctgct ctggatgatg
     4261 gccaggtatt ccacgggcac ggttttctcg atgttctcga acttctgata cttgtagtgg
     4321 ccggttttct ggttccgctg cttgttaatc ttgatcactt cattggtgat cttaaagaac
     4381 accttggagt ttttcacgaa cttgttcagg aactcgccgt agtagatatt cttcagcagg
     4441 tagatctggg cgtccttctc ttctttgtcc ttgggcacgc tccaaaaaaa cttcagggta
     4501 ttccgcaggt cctcaatctt gttgtacagc ttggtgaagc tgggcacgaa ggggatgttt
     4561 ttgttcacga agttgaactt ggtattcttc aggtacttga tgatcacatc cttctcgtag
     4621 tagttgaaca cgttggcgct gttcagctgc ttgaagattt tcagcttcag cttcttttcg
     4681 ttgatttcgt tctgaaacat cttcttggag atctcgctgg gggcgatatt cttgaaggcg
     4741 aagatgtcct tgccttccag ttccaggttg aagtgcacga tgccgtgtct gatgctgctg
     4801 atggcctcgt cgatgttggc gaagaagtcc tcgatctcgt tcttgttgtc catgttgaag
     4861 tcgtagctgt agaacatctt cagattttct ttcacttcgt tctgcttgtt ctcattgtag
     4921 atcttgtcca cctcgccgga cacgtatttc tcttcgccct tgttgttctt cacggtcttg
     4981 ccccgcatcc ggccggtgat atcgttctcg ttctcggttt ccaggatgtt cctcaggctg
     5041 aagtaggcca cgctggacac gccgatgatg tttctcagga aggcctcgtt ctgccggttc
     5101 cgggcgataa agtcggaggt ggcgatctcg cccacttgca gatagtagtt gtacttgccg
     5161 cagttccgca cgtaggtgtc cagcttgttc agcagtttgt tttcgatcag ctttttcaga
     5221 ttctggtact cgaagatccg cttgatctta tcgttgctga tgttgctcag ccgcttgtac
     5281 acgtagtttt tcagcagctg ggacatctcg atttccacga agtggcagaa ggcgtactta
     5341 atattcttgt cgttcagttc ctctttgtcc aggtagtact tgtagaacac ctggcttttc
     5401 ttcagctcag acatgtcggg gatcttctca atcagctctt tgatgttgtt cacgttctgg
     5461 atctcttcgt agataatctt ggcgaagttc tctttgtcgt tcttccggcc gatgatcttg
     5521 tgatagtact cgcggatctt gtacttctcg tgcttcttgc tgttctcgat caggaaaaac
     5581 agtttctcga tatcctcttt cttatacagc ttgtcgaagg cttcctgcac gttgttgatg
     5641 tagtcgttgc gcttggcgct ctctctgtag tagtcgtaga tgatgttccg cttgctcttg
     5701 ccgcccactt tttccacgtt gttctcgttg atcttctggt agttggcctt gttctcttcg
     5761 aagctgtact tcaggctgtt gatcttgttc agcttggctt ccacgtcctt ccgaaagatt
     5821 tccagttcct cagagttcac gtcctcgttc agcaggatct tcttcagcac ggagaagctg
     5881 ttcttgtttt tcaggtcgta ctcgctgatg tcctcttcgc tatagttctt gtcctgcacg
     5941 gcgttctttt ctttccggtt cttcagatac agcacgctgt ccttcaggtg cagcaccttg
     6001 ttgctaaaga acttcttcag gttctctctt ctgatccggt tctcttcctc ggaggcgttg
     6061 tcggggttct tgatgtagat gtccagccgg atgctcagca gctcgctcag tctctcgctg
     6121 gtccggtttt cctcgctggt ggacttcacg agcttgccct cttcgatgta cttcttgtgg
     6181 ctgatgccgt cgaccttggt cactttcatg gatccaccaa tctgttctct gtgagcctca
     6241 ataatatcgt tatcctccat gtccaaatct tcaggggtct gatcagcttg gattctaata
     6301 ccgtcgtaca agaatcttaa ggagtccatt tccttaccct gtcttttagc gaacgcttcc
     6361 atcagccttc ttaaaggagt ggtctttttg atcttgaaga agatctctga agatccatcg
     6421 gacaccttta aattgatgtg agtctcaggc ttgacttctg gcttgacctc tggcttagct
     6481 tcttgattga cttctgagtc cgacattttc tcaaactgag ggtgtgacca tgcactacca
     6541 cctgaaccac caccgctacc accacctttt tcaaactgcg gatggctcca gctagccata
     6601 tggctgccgc gcggcaccag gccgctgctg tgatgatgat gatgatggct gctgcccatg
     6661 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
     6721 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
     6781 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
     6841 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
     6901 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
     6961 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
     7021 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
     7081 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     7141 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     7201 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     7261 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     7321 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     7381 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     7441 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     7501 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     7561 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     7621 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     7681 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     7741 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     7801 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     7861 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     7921 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     7981 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     8041 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     8101 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8161 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     8221 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
     8281 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
     8341 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
     8401 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
     8461 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
     8521 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg
     8581 cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca
     8641 tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac
     8701 agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga ccgctgatcg
     8761 tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg attgtaggcg
     8821 ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct
     8881 cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa gaattggagc
     8941 caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat
     9001 cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc
     9061 acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
     9121 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
     9181 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
     9241 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
     9301 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
     9361 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
     9421 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
     9481 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
     9541 aa
//