LOCUS       sequence-337610-        8236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of NLS-RfxCas13d-NLS 6His.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8236)
  AUTHORS   Mendez-Mancilla A, Wessels HH, Legut M, Kadina A, Mabuchi M, Walker 
            J, Robb GB, Holden K, Sanjana NE
  TITLE     Chemically modified guide RNAs enhance CRISPR-Cas13 knockdown in 
            human cells.
  JOURNAL   Cell Chem Biol. 2022 Feb 17;29(2):321-327.e4. doi: 
            10.1016/j.chembiol.2021.07.011. Epub 2021 Aug 2.
  PUBMED    34343484
REFERENCE   2  (bases 1 to 8236)
  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: 
            "10.1016/j.chembiol.2021.07.011"; journalName: "Cell Chem Biol"; 
            date: "2022-02-17- 17"; volume: "29"; issue: "2"; pages: 
            "321-327.e4"
COMMENT     Sequence Label: pET28-MKIEE-NLS-RfxCas13d-NLS-HA-6His
FEATURES             Location/Qualifiers
     source          1..8236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(35..1117)
                     /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     1079..1098
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1118..1195)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1401..1420
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1504..1523
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1504..1522
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1523..1547
                     /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)."
     RBS             1562..1584
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1611..1631
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1641..4538
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             4548..4568
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4578..4604
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4605..4622
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4674..4692)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4688..4735
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4772..5227
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4859..4878)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5069..5090
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5319..6134)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5319..6134)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6042..6061
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6126..6145
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6256..6844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6745..6764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6998..7015
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7030..7169
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7095..7114)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7255..7277
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7271..7462)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
       61 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      121 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
      181 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
      241 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
      301 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
      361 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
      421 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
      481 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
      541 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
      601 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
      661 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
      721 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
      781 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
      841 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
      901 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
      961 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
     1021 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
     1081 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
     1141 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
     1201 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
     1261 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
     1321 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
     1381 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
     1441 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
     1501 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
     1561 ttttgtttaa ctttaagaag gagatataca tgaaaatcga agaaatgagc cccaagaaga
     1621 agagaaaggt ggaggccagc atcgaaaaaa aaaagtcctt cgccaagggc atgggcgtga
     1681 agtccacact cgtgtccggc tccaaagtgt acatgacaac cttcgccgaa ggcagcgacg
     1741 ccaggctgga aaagatcgtg gagggcgaca gcatcaggag cgtgaatgag ggcgaggcct
     1801 tcagcgctga aatggccgat aaaaacgccg gctataagat cggcaacgcc aaattcagcc
     1861 atcctaaggg ctacgccgtg gtggctaaca accctctgta tacaggaccc gtccagcagg
     1921 atatgctcgg cctgaaggaa actctggaaa agaggtactt cggcgagagc gctgatggca
     1981 atgacaatat ttgtatccag gtgatccata acatcctgga cattgaaaaa atcctcgccg
     2041 aatacattac caacgccgcc tacgccgtca acaatatctc cggcctggat aaggacatta
     2101 ttggattcgg caagttctcc acagtgtata cctacgacga attcaaagac cccgagcacc
     2161 atagggccgc tttcaacaat aacgataagc tcatcaacgc catcaaggcc cagtatgacg
     2221 agttcgacaa cttcctcgat aaccccagac tcggctattt cggccaggcc tttttcagca
     2281 aggagggcag aaattacatc atcaattacg gcaacgaatg ctatgacatt ctggccctcc
     2341 tgagcggact gaggcactgg gtggtccata acaacgaaga agagtccagg atctccagga
     2401 cctggctcta caacctcgat aagaacctcg acaacgaata catctccacc ctcaactacc
     2461 tctacgacag gatcaccaat gagctgacca actccttctc caagaactcc gccgccaacg
     2521 tgaactatat tgccgaaact ctgggaatca accctgccga attcgccgaa caatatttca
     2581 gattcagcat tatgaaagag cagaaaaacc tcggattcaa tatcaccaag ctcagggaag
     2641 tgatgctgga caggaaggat atgtccgaga tcaggaaaaa tcataaggtg ttcgactcca
     2701 tcaggaccaa ggtctacacc atgatggact ttgtgattta taggtattac atcgaagagg
     2761 atgccaaggt ggctgccgcc aataagtccc tccccgataa tgagaagtcc ctgagcgaga
     2821 aggatatctt tgtgattaac ctgaggggct ccttcaacga cgaccagaag gatgccctct
     2881 actacgatga agctaataga atttggagaa agctcgaaaa tatcatgcac aacatcaagg
     2941 aatttagggg aaacaagaca agagagtata agaagaagga cgcccctaga ctgcccagaa
     3001 tcctgcccgc tggccgtgat gtttccgcct tcagcaaact catgtatgcc ctgaccatgt
     3061 tcctggatgg caaggagatc aacgacctcc tgaccaccct gattaataaa ttcgataaca
     3121 tccagagctt cctgaaggtg atgcctctca tcggagtcaa cgctaagttc gtggaggaat
     3181 acgccttttt caaagactcc gccaagatcg ccgatgagct gaggctgatc aagtccttcg
     3241 ctagaatggg agaacctatt gccgatgcca ggagggccat gtatatcgac gccatccgta
     3301 ttttaggaac caacctgtcc tatgatgagc tcaaggccct cgccgacacc ttttccctgg
     3361 acgagaacgg aaacaagctc aagaaaggca agcacggcat gagaaatttc attattaata
     3421 acgtgatcag caataaaagg ttccactacc tgatcagata cggtgatcct gcccacctcc
     3481 atgagatcgc caaaaacgag gccgtggtga agttcgtgct cggcaggatc gctgacatcc
     3541 agaaaaaaca gggccagaac ggcaagaacc agatcgacag gtactacgaa acttgtatcg
     3601 gaaaggataa gggcaagagc gtgagcgaaa aggtggacgc tctcacaaag atcatcaccg
     3661 gaatgaacta cgaccaattc gacaagaaaa ggagcgtcat tgaggacacc ggcagggaaa
     3721 acgccgagag ggagaagttt aaaaagatca tcagcctgta cctcaccgtg atctaccaca
     3781 tcctcaagaa tattgtcaat atcaacgcca ggtacgtcat cggattccat tgcgtcgagc
     3841 gtgatgctca actgtacaag gagaaaggct acgacatcaa tctcaagaaa ctggaagaga
     3901 agggattcag ctccgtcacc aagctctgcg ctggcattga tgaaactgcc cccgataaga
     3961 gaaaggacgt ggaaaaggag atggctgaaa gagccaagga gagcattgac agcctcgaga
     4021 gcgccaaccc caagctgtat gccaattaca tcaaatacag cgacgagaag aaagccgagg
     4081 agttcaccag gcagattaac agggagaagg ccaaaaccgc cctgaacgcc tacctgagga
     4141 acaccaagtg gaatgtgatc atcagggagg acctcctgag aattgacaac aagacatgta
     4201 ccctgttcag aaacaaggcc gtccacctgg aagtggccag gtatgtccac gcctatatca
     4261 acgacattgc cgaggtcaat tcctacttcc aactgtacca ttacatcatg cagagaatta
     4321 tcatgaatga gaggtacgag aaaagcagcg gaaaggtgtc cgagtacttc gacgctgtga
     4381 atgacgagaa gaagtacaac gataggctcc tgaaactgct gtgtgtgcct ttcggctact
     4441 gtatccccag gtttaagaac ctgagcatcg aggccctgtt cgataggaac gaggccgcca
     4501 agttcgacaa ggagaaaaag aaggtgtccg gcaattccgg atccggacct aagaaaaaga
     4561 ggaaggtggc ggccgcttac ccatacgatg ttccagatta cgctcatcac catcaccatc
     4621 attaaatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     4681 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     4741 aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     4801 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4861 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4921 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4981 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5041 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5101 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5161 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     5221 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     5281 aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc atcgagcatc
     5341 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     5401 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     5461 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     5521 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     5581 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     5641 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     5701 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     5761 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     5821 gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     5881 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     5941 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     6001 ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     6061 ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga cgtttcccgt
     6121 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     6181 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6241 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6361 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6421 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6481 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6541 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6601 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6661 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6721 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6781 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6841 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6901 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6961 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7021 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcaa
     7081 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     7141 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     7201 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     7261 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     7321 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     7381 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     7441 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     7501 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     7561 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     7621 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     7681 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     7741 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     7801 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     7861 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     7921 gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg
     7981 cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca
     8041 ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg
     8101 ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag
     8161 tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc
     8221 gagatcccgg tgccta
//
