LOCUS       sequence-329649-       10206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of LbCas12a using CL7/Im7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10206)
  AUTHORS   Nguyen LT, Gurijala J, Rananaware SR, Pizzano BLM, Stone BT, Jain PK
  TITLE     CRISPR-ENHANCE: An enhanced nucleic acid detection platform using 
            Cas12a.
  JOURNAL   Methods. 2021 Feb 9. pii: S1046-2023(21)00025-6. doi: 
            10.1016/j.ymeth.2021.02.001.
  PUBMED    33577982
REFERENCE   2  (bases 1 to 10206)
  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"; journalName: "Methods. 
            2021 Feb 9. pii: S1046-2023(21)00025-6. doi: 
            10.1016/j.ymeth.2021.02.001."
COMMENT     Sequence Label: CL7-LbCas12a
FEATURES             Location/Qualifiers
     source          1..10206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     960..979
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(999..1076)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1282..1301
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1385..1404
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1385..1403
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1404..1428
                     /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)."
     CDS             1472..1798
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYSGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             2213..2236
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2261..2284
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2288..2311
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             2663..6346
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     CDS             6350..6376
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6356..6382
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             6395..6415
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6422..6442
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6455..6472
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6525..6543)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6539..6586
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6623..7078
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6710..6729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6920..6941
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7170..7985)
                     /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(7170..7985)
                     /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     7893..7912
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7977..7996
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8107..8695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8596..8615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8849..8866
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8881..9020
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8946..8965)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9106..9128
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9122..9313)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(10122..10206,1..998))
                     /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 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
       61 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
      121 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
      181 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
      241 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
      301 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
      361 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
      421 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
      481 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
      541 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
      601 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
      661 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
      721 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
      781 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
      841 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
      901 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
      961 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     1021 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     1081 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     1141 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     1201 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     1261 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     1321 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     1381 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     1441 attttgttta accttaagaa ggagatatac catgagcgat aaaattattc acctgactga
     1501 cgacagtttt gacacggatg tactcaaagc ggacggggcg atcctcgtcg atttctgggc
     1561 agagtggtgt ggtccgtgca aaatgatcgc cccgattctg gatgaaatcg ctgacgaata
     1621 ttctggcaaa ctgaccgttg caaaactgaa catcgatcaa aaccctggca ctgcgccgaa
     1681 atatggcatc cgtggtatcc cgactctgct gctgttcaaa aacggtgaag tggcggcaac
     1741 caaagtgggt gcactgtcta aaggtcagtt gaaagagttc ctcgacgcta acctggcagg
     1801 tagtggtacg aattctgaac cgggtgaagc aactggtgag ggtgaaccag ttaacaacaa
     1861 atggctggat aacgcaggta aagatctggg ttctccagtt cctgatcgca ttgctgataa
     1921 gctgcgcgat caagaatttg aatcctttga cgatttccgt gaaacctttt gggaagaagt
     1981 ctctaaagac ccggaactga gcaaacagtt ctcccgcaac aacaacgacc gcatgaaagt
     2041 gggcaaagct ccgaaaaccc gtactcaaga cgtttctggc aaacgcacta gctttgaact
     2101 gaaccatgag aaaccgattg aagaaaacgg cggtgtttat gatatggaca acatctccgt
     2161 ggttacgccg aaacgtaaca tcgatatcga aaaccatggt gacaatacgt ctctggaagt
     2221 actgtttcaa ggtcctagtt ctaacacctc ttctaacggc ttagaggttc tgtttcaggg
     2281 tccaggtcat catcaccatc accatcatca caacgagacg tctggttctc tgcaagattc
     2341 tgaagttaat caggaagcta aaccggaagt aaaaccagag gttaaaccag aaactcacat
     2401 taacctgaaa gtgagcgatg gttcttccga aatctttttc aagattaaaa agactactcc
     2461 gctgcgtcgt ctgatggaag cattcgcgaa acgccaaggt aaagaaatgg attctctgcg
     2521 ctttctgtat gacggcattc gcattcaggc tgaccaagca cctgaggatc tggacatgga
     2581 agacaatgac attatcgaag ctcatcgtga acagattggt ggttctggta ccctgcctga
     2641 aactggttct agcgaagctt ccatgagcaa actggagaag ttcacgaact gttattctct
     2701 gtcgaaaacc ttacgtttta aggccattcc ggtaggtaaa acgcaagaaa atatcgacaa
     2761 taaacgtctc ctggttgaag atgaaaagcg tgccgaagat tataaaggtg ttaaaaagct
     2821 gcttgatcgc tattacctgt cttttatcaa cgatgttctg cattccatca aattgaaaaa
     2881 cttgaataat tacatctcgt tatttcgcaa aaaaacccgt accgaaaaag aaaacaagga
     2941 gctggaaaac cttgaaatta atctgcgtaa agaaatcgcg aaagcattta aaggtaatga
     3001 aggatataaa tctctgttta aaaaagacat tattgaaacc atcctgcctg agttcttgga
     3061 tgataaagat gaaattgccc tggttaatag ctttaatggc tttacgactg catttaccgg
     3121 cttttttgat aaccgtgaaa acatgttcag cgaagaagcg aaatcgactt ccattgcctt
     3181 ccgttgcatt aatgaaaacc tgacccgcta catttctaat atggacatct ttgaaaaagt
     3241 tgatgccatt ttcgacaaac atgaagttca agaaatcaaa gaaaagattt taaactccga
     3301 ttatgatgtt gaagatttct ttgagggaga atttttcaat tttgttctca cccaagaggg
     3361 tattgatgtc tataacgcca ttatcggagg atttgttacg gaatcaggcg aaaagattaa
     3421 aggactgaat gaatacatta atttatacaa tcagaaaacc aaacagaaat tgccaaaatt
     3481 caaaccactg tataaacagg tattaagcga ccgtgaatcc ttgtcctttt atggcgaagg
     3541 ctatacaagc gatgaagagg tgctcgaagt attccgtaat actttgaaca aaaactcgga
     3601 aatcttctcc agcattaaaa aactggaaaa attgtttaaa aatttcgatg aatatagtag
     3661 cgcaggcatc ttcgtcaaaa atggccctgc gatttcaacc atttctaaag acattttcgg
     3721 tgaatggaac gtgatccgcg ataaatggaa cgcggagtat gacgacattc atctgaaaaa
     3781 aaaagccgtt gttaccgaaa agtatgaaga tgaccgccgc aagagcttta aaaaaattgg
     3841 tagtttctca ctggaacagt tacaagaata tgcagacgcc gatctgtcgg ttgtggaaaa
     3901 gttaaaagag attattatcc agaaagtcga tgaaatttat aaagtgtacg gctccagcga
     3961 aaagctcttt gatgccgact ttgttttaga aaaatcgctg aaaaaaaacg atgccgtggt
     4021 ggccatcatg aaagatctgt tggattcggt caaatcgttc gaaaactata tcaaggcgtt
     4081 ctttggcgaa gggaaagaga ccaatcgtga cgaatcgttc tatggcgatt ttgtgttggc
     4141 gtatgatatt cttctgaaag tagatcacat ctatgatgct atccgcaact acgtgaccca
     4201 gaaaccgtat tcgaaggata aatttaagct gtattttcaa aaccctcagt ttatgggtgg
     4261 ctgggataaa gataaagaaa ccgactaccg cgcgacgatc ttgcgctatg gctctaagta
     4321 ctatttagcc atcatggata aaaagtatgc aaaatgcctc cagaaaatcg ataaggatga
     4381 tgtcaacggg aactacgaaa aaatcaacta caaactcttg cctggcccga ataaaatgct
     4441 gccgaaagtc tttttttcca aaaaatggat ggcctattat aacccttccg aagatattca
     4501 gaagatttac aagaacggta cctttaaaaa aggtgacatg tttaatctga atgactgtca
     4561 caagcttatt gatttcttca aagatagtat ttctcgctac cctaaatggt caaacgcgta
     4621 cgattttaat ttctcagaaa cagagaaata caaagatatt gcggggtttt atcgcgaagt
     4681 cgaagaacaa gggtataagg tgtcgttcga atcggcttcc aaaaaggaag tggataaact
     4741 tgttgaagaa ggcaaactct atatgttcca gatctacaac aaagatttct ccgacaagag
     4801 ccatggcacg ccgaacttac ataccatgta ctttaaatta ctgttcgatg aaaataatca
     4861 tggtcagatt cgcttgagcg gtggcgccga actgtttatg cgccgcgcat cattgaaaaa
     4921 ggaagaactt gttgtgcacc cagcaaacag tccgattgca aacaaaaatc cggataaccc
     4981 gaagaagact accaccttaa gctatgatgt atataaagat aaacgtttta gcgaagatca
     5041 gtatgaactc cacattccga tcgcgattaa caaatgtcct aagaacatct ttaaaatcaa
     5101 taccgaggtt cgtgtactgc tcaaacacga cgacaacccc tatgttattg gcattgaccg
     5161 cggcgaacgc aacctgctgt atattgtcgt tgtggatggc aaaggtaaca ttgttgaaca
     5221 gtatagcctt aatgaaatca tcaataattt caacggcatt cgtattaaaa cggactatca
     5281 tagcctgctg gacaaaaaag agaaagaacg cttcgaagcc cgtcagaatt ggacctctat
     5341 cgaaaatatc aaggaactga aagctgggta catttctcaa gtggtccaca agatttgcga
     5401 actggtggaa aagtatgacg cggtgatcgc actggaggat ctgaattcgg gctttaaaaa
     5461 tagccgtgtg aaggtggaga aacaggttta ccaaaagttc gaaaaaatgc tgattgacaa
     5521 acttaattac atggtggaca aaaaatctaa tccatgtgcg accggcgggg ctctcaaagg
     5581 ctatcagatt acgaataaat tcgaaagctt caaatccatg tctacccaga acggatttat
     5641 cttttacatc cccgcctggc tgacttccaa aattgatccg agtaccggtt ttgttaatct
     5701 gctgaaaacc aaatatacgt cgatcgccga ttcaaaaaaa ttcattagca gcttcgaccg
     5761 tattatgtac gtcccggaag aagacctgtt cgaattcgcc cttgactata aaaatttctc
     5821 gcgtacggat gccgattata tcaaaaaatg gaaactgtac agctacggca accgcatccg
     5881 catttttcgc aacccaaaga agaacaacgt atttgactgg gaggaagttt gcttgacgag
     5941 cgcttacaaa gagctcttta acaaatatgg gatcaattat cagcaagggg acatccgtgc
     6001 gttgctgtgt gagcagtcag ataaagcgtt ctacagctct ttcatggctt taatgtcgct
     6061 gatgttgcag atgcgcaatt cgatcactgg ccgcaccgat gtggatttcc tgattagccc
     6121 ggtcaaaaat agcgatggca ttttttacga ttcacgtaac tatgaagccc aggaaaacgc
     6181 tattctgccg aagaacgctg atgcgaacgg cgcgtacaat attgcccgta aggtactgtg
     6241 ggcaattggt cagttcaaga aagcggaaga tgaaaagttg gataaggtca aaattgcgat
     6301 ctcgaataaa gaatggctgg aatatgcgca gaccagcgtg aaacatgcct atccctatga
     6361 cgtgcccgat tatgccagcc tgggcagcgg ctcccccaag aaaaaacgca aggtggaaga
     6421 tcctaagaaa aagcggaaag tggactaact cgagcaccac caccaccacc actgagatcc
     6481 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     6541 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     6601 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     6661 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     6721 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6781 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6841 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6901 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6961 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     7021 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     7081 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     7141 caaatatgta tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac
     7201 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat
     7261 gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg
     7321 attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta
     7381 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc
     7441 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca
     7501 tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg
     7561 ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca
     7621 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg
     7681 gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc
     7741 ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg
     7801 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat
     7861 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa
     7921 tcagcatcca tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg
     7981 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa
     8041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     8101 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     8161 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     8221 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     8281 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     8341 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     8401 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     8461 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     8521 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     8581 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     8641 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     8701 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     8761 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     8821 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     8881 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     8941 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     9001 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     9061 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     9121 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     9181 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     9241 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     9301 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     9361 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     9421 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     9481 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     9541 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     9601 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     9661 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     9721 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     9781 gggccgccat gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag
     9841 tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca
     9901 tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct
     9961 gtcctacgag ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc
    10021 gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg
    10081 gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt
    10141 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
    10201 tgcgta
//
