LOCUS       sequence-372771-        9491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET21a protein expression vector for 1N/2CxNLS-cMyc-NLP-cMyc 
            enAspCas12a in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9491)
  AUTHORS   Luk K, Liu P, Zeng J, Wang Y, Maitland SA, Idrizi F, Ponnienselvan 
            K, Iyer S, Zhu LJ, Luban J, Bauer DE, Wolfe SA
  TITLE     Optimization of Nuclear Localization Signal Composition Improves 
            CRISPR-Cas12a Editing Rates in Human Primary Cells.
  JOURNAL   GEN Biotechnol. 2022 Jun;1(3):271-284. doi: 
            10.1089/genbio.2022.0003. Epub 2022 Jun 14.
  PUBMED    38405215
REFERENCE   2  (bases 1 to 9491)
  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: 
            "10.1089/genbio.2022.0003"; journalName: "GEN Biotechnol"; date: 
            "2022-06"; volume: "1"; issue: "3"; pages: "271-284"
COMMENT     Sequence Label: 1N/2CxNLS-cMyc-NLP-cMyc enAspCas12a
FEATURES             Location/Qualifiers
     source          1..9491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     582..601
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(621..698)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     904..923
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1007..1026
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1007..1025
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1026..1050
                     /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             1065..1087
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1098..1124
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1143..5060
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     CDS             5061..5108
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5115..5195
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             5196..5222
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5229..5246
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5299..5317)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5313..5360
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5397..5852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5484..5503)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5694..5715
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5878..5982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5983..6051,6052..6843)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6201..6220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7014..7602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7503..7522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7756..7773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7788..7927
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7853..7872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8013..8035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8029..8220)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(9029..9491,1..620))
                     /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"
ORIGIN
        1 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
       61 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
      121 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
      181 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
      241 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
      301 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
      361 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
      421 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
      481 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
      541 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
      601 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
      661 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
      721 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
      781 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
      841 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
      901 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
      961 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
     1021 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
     1081 aaggagatat acatatgccc gccgccaagc gcgtgaagct ggacggcggt agcggcggag
     1141 gcacacagtt cgagggcttt accaacctgt atcaggtgag caagacactg cggtttgagc
     1201 tgatcccaca gggcaagacc ctgaagcaca tccaggagca gggcttcatc gaggaggaca
     1261 aggcccgcaa tgatcactac aaggagctga agcccatcat cgatcggatc tacaagacct
     1321 atgccgacca gtgcctgcag ctggtgcagc tggattggga gaacctgagc gccgccatcg
     1381 actcctatag aaaggagaaa accgaggaga caaggaacgc cctgatcgag gagcaggcca
     1441 catatcgcaa tgccatccac gactacttca tcggccggac agacaacctg accgatgcca
     1501 tcaataagag acacgccgag atctacaagg gcctgttcaa ggccgagctg tttaatggca
     1561 aggtgctgaa gcagctgggc accgtgacca caaccgagca cgagaacgcc ctgctgcgga
     1621 gcttcgacaa gtttacaacc tacttctccg gcttttatag aaacaggaag aacgtgttca
     1681 gcgccgagga tatcagcaca gccatcccac accgcatcgt gcaggacaac ttccccaagt
     1741 ttaaggagaa ttgtcacatc ttcacacgcc tgatcaccgc cgtgcccagc ctgcgggagc
     1801 actttgagaa cgtgaagaag gccatcggca tcttcgtgag cacctccatc gaggaggtgt
     1861 tttccttccc tttttataac cagctgctga cacagaccca gatcgacctg tataaccagc
     1921 tgctgggagg aatctctcgg gaggcaggca ccgagaagat caagggcctg aacgaggtgc
     1981 tgaatctggc catccagaag aatgatgaga cagcccacat catcgcctcc ctgccacaca
     2041 gattcatccc cctgtttaag cagatcctgt ccgataggaa caccctgtct ttcatcctgg
     2101 aggagtttaa gagcgacgag gaagtgatcc agtccttctg caagtacaag acactgctga
     2161 gaaacgagaa cgtgctggag acagccgagg ccctgtttaa cgagctgaac agcatcgacc
     2221 tgacacacat cttcatcagc cacaagaagc tggagacaat cagcagcgcc ctgtgcgacc
     2281 actgggatac actgaggaat gccctgtatg agcggagaat ctccgagctg acaggcaaga
     2341 tcaccaagtc tgccaaggag aaggtgcagc gcagcctgaa gcacgaggat atcaacctgc
     2401 aggagatcat ctctgccgca ggcaaggagc tgagcgaggc cttcaagcag aaaaccagcg
     2461 agatcctgtc ccacgcacac gccgccctgg atcagccact gcctacaacc ctgaagaagc
     2521 aggaggagaa ggagatcctg aagtctcagc tggacagcct gctgggcctg taccacctgc
     2581 tggactggtt tgccgtggat gagtccaacg aggtggaccc cgagttctct gcccggctga
     2641 ccggcatcaa gctggagatg gagccttctc tgagcttcta caacaaggcc agaaattatg
     2701 ccaccaagaa gccctactcc gtggagaagt tcaagctgaa ctttcagatg cctacactgg
     2761 ccagaggctg ggacgtgaat agagagaaga acaatggcgc catcctgttt gtgaagaacg
     2821 gcctgtacta tctgggcatc atgccaaagc agaagggcag gtataaggcc ctgagcttcg
     2881 agcccacaga gaaaaccagc gagggctttg ataagatgta ctatgactac ttccctgatg
     2941 ccgccaagat gatcccaaag tgcagcaccc agctgaaggc cgtgacagcc cactttcaga
     3001 cccacacaac ccccatcctg ctgtccaaca atttcatcga gcctctggag atcacaaagg
     3061 agatctacga cctgaacaat cctgagaagg agccaaagaa gtttcagaca gcctacgcca
     3121 agaaaaccgg cgaccagaag ggctacagag aggccctgtg caagtggatc gacttcacaa
     3181 gggattttct gtccaagtat accaagacaa cctctatcga tctgtctagc ctgcggccat
     3241 cctctcagta taaggacctg ggcgagtact atgccgagct gaatcccctg ctgtaccaca
     3301 tcagcttcca gagaatcgcc gagaaggaga tcatggatgc cgtggagaca ggcaagctgt
     3361 acctgttcca gatctataac aaggactttg ccaagggcca ccacggcaag cctaatctgc
     3421 acacactgta ttggaccggc ctgttttctc cagagaacct ggccaagaca agcatcaagc
     3481 tgaatggcca ggccgagctg ttctaccgcc ctaagtccag gatgaagagg atggcacacc
     3541 ggctgggaga gaagatgctg aacaagaagc tgaaggatca gaaaacccca atccccgaca
     3601 ccctgtacca ggagctgtac gactatgtga atcacagact gtcccacgac ctgtctgatg
     3661 aggccagggc cctgctgccc aacgtgatca ccaaggaggt gtctcacgag atcatcaagg
     3721 ataggcgctt taccagcgac aagttctttt tccacgtgcc tatcacactg aactatcagg
     3781 ccgccaattc cccatctaag ttcaaccaga gggtgaatgc ctacctgaag gagcaccccg
     3841 agacacctat catcggcatc gatcggggcg agagaaacct gatctatatc acagtgatcg
     3901 actccaccgg caagatcctg gagcagcgga gcctgaacac catccagcag tttgattacc
     3961 agaagaagct ggacaacagg gagaaggaga gggtggcagc aaggcaggcc tggtctgtgg
     4021 tgggcacaat caaggatctg aagcagggct atctgagcca ggtcatccac gagatcgtgg
     4081 acctgatgat ccactaccag gccgtggtgg tgctggagaa cctgaatttc ggctttaaga
     4141 gcaagaggac cggcatcgcc gagaaggccg tgtaccagca gttcgagaag atgctgatcg
     4201 ataagctgaa ttgcctggtg ctgaaggact atccagcaga gaaagtggga ggcgtgctga
     4261 acccatacca gctgacagac cagttcacct cctttgccaa gatgggcacc cagtctggct
     4321 tcctgtttta cgtgcctgcc ccatatacat ctaagatcga tcccctgacc ggcttcgtgg
     4381 accccttcgt gtggaaaacc atcaagaatc acgagagccg caagcacttc ctggagggct
     4441 tcgactttct gcactacgac gtgaaaaccg gcgacttcat cctgcacttt aagatgaaca
     4501 gaaatctgtc cttccagagg ggcctgcccg gctttatgcc tgcatgggat atcgtgttcg
     4561 agaagaacga gacacagttt gacgccaagg gcaccccttt catcgccggc aagagaatcg
     4621 tgccagtgat cgagaatcac agattcaccg gcagataccg ggacctgtat cctgccaacg
     4681 agctgatcgc cctgctggag gagaagggca tcgtgttcag ggatggctcc aacatcctgc
     4741 caaagctgct ggagaatgac gattctcacg ccatcgacac gatggtggcc ctgatccgca
     4801 gcgtgctgca gatgcggaac tccaatgccg ccacaggcga ggactatatc aacagccccg
     4861 tgcgcgatct gaatggcgtg tgcttcgact cccggtttca gaacccagag tggccaatgg
     4921 acgccgatgc caatggcgcc taccacatcg ccctgaaggg ccagctgctg ctgaatcacc
     4981 tgaaggagag caaggatctg aagctgcaga acggcatctc caatcaggac tggctggcct
     5041 acatccagga gctgcgcaac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa
     5101 agaaaaaggg atcctaccca tacgatgttc cagattacgc ttatccctac gacgtgcctg
     5161 attatgcata cccatatgat gtccccgact atgccccggc agctaagagg gtgaaactgg
     5221 atctcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     5281 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     5341 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     5401 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5461 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5521 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5581 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5641 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5701 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5761 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5821 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     5881 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5941 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6001 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6061 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6121 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6181 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6241 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6301 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6361 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6421 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6481 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     6541 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6601 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6661 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6721 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6781 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6841 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6901 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6961 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7021 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7081 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7141 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     7201 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7261 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7321 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7381 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7441 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7561 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7621 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     7681 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     7741 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     7801 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     7861 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     7921 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     7981 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     8041 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     8101 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     8161 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     8221 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     8281 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     8341 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     8401 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     8461 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     8521 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     8581 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     8641 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc
     8701 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     8761 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     8821 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     8881 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     8941 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     9001 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     9061 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     9121 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     9181 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     9241 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     9301 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     9361 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     9421 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     9481 gaatttgatt g
//
