LOCUS       sequence-364005-       10963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-MBP tagged BsCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10963)
  AUTHORS   Nguyen LT,  Macaluso NC,  Jain PK
  TITLE     A Combinatorial Approach towards Adaptability of 22 Functional 
            Cas12a Orthologs for Nucleic Acid Detection in Clinical Samples
  JOURNAL   medRxiv 2021.07.21
REFERENCE   2  (bases 1 to 10963)
  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: "medRxiv 
            2021.07.21"
COMMENT     Sequence Label: 6xHis-MBP-huBsCas12a
FEATURES             Location/Qualifiers
     source          1..10963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1242..1433
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1427..1449)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1535..1674
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1590..1609
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1689..1706)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1860..2448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1940..1959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2559..2578)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2570..3385
                     /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             2570..3385
                     /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     complement(2643..2662)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3478..3933)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3615..3636)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3827..3846
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3970..4017)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4013..4031
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4084..4101)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(join(4102..4812,4813..4815,4816..4818))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(4102..4812,4813..4815,4816..4818))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4144..4165)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4493..4512
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4752..4773
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(4819..4872)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(4894..4983)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(4990..5010)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8641..8661)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(8668..8688)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(8701..9801)
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     complement(8704..8727)
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             complement(9811..9828)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             9847..9869
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    9884..9908
                     /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(9908..9927)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9909..9927)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10011..10030)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        10236..10313
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(10314..10963,1..433)
                     /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(10333..10352)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggtat
       61 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
      121 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
      181 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
      241 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
      301 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
      361 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
      421 aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat
      481 ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca
      541 tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta ggacaggtgc
      601 cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg acgatgatcg
      661 gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc gtcactggtc
      721 ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg gccccacggg
      781 tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg
      841 gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt
      901 ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa
      961 cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc
     1021 taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt
     1081 ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc
     1141 gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca
     1201 ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa
     1261 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca
     1321 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg
     1381 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     1441 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     1501 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     1561 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     1621 ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
     1681 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1741 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1801 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1861 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1921 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1981 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2041 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2101 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2161 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2221 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2281 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2341 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2401 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2461 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2521 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     2581 tcaacgggaa acgtcttgct ctaggccgcg attaaattcc aacatggatg ctgatttata
     2641 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     2701 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     2761 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2821 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa
     2881 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaacattg ttgatgcgct
     2941 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3001 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3061 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3121 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     3181 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     3241 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     3301 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     3361 tttgatgctc gatgagtttt tctaagaatt aattcatgag cggatacata tttgaatgta
     3421 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgaaa
     3481 ttgtaaacgt tactagtttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3541 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3601 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3661 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3721 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3781 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3841 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3901 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccaatccgg atatagttcc
     3961 tcctttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta
     4021 ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc
     4081 tcagtggtgg tggtggtggt gcttgtacag ctcgtccatg ccgagagtga tcccggcggc
     4141 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
     4201 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     4261 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     4321 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     4381 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     4441 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     4501 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     4561 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta
     4621 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     4681 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     4741 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     4801 ctcgcccttg ctcaccattg ggccgggatt ctcctccacg tcaccgcatg ttagaagact
     4861 tcctctgccc tctccgccgc cggacctgga tccggcgtag tctgggacgt cataagggta
     4921 ggagcctgca tagtcaggga catcgtatgg gtatccggca taatctggaa catcataagg
     4981 gtatctagag actttcctct tcttctttgg gctagccagc aggtgcagct gggcgtactt
     5041 cagccactcg gcgttggtca tgctcaggtt caccttctcg ccctcgtcct tctggcggat
     5101 ctgctccagc acccacaggc ccttgcgggc gatgttgtag gcgccgttgg cgtcggcgtc
     5161 cttgggcatg gtgccctcgc tgaactcgct gcagaagaac tcgccgcggt cgttcttcac
     5221 ggggctgatg atgtagtcgc gggtgccgtc gctggtgctg ttgcgcatct gcagggtctg
     5281 gtgcagcagg cggaacaggc gcttgtagaa gtcggcctcg gccttgctga tgatgatctc
     5341 cttgatgtcc tcgccgttct cgtaggggat gcggtactgc ttgaacaggc ccttgatctc
     5401 gtcggtcacg ttgatcacct tctcgtcgaa caggttgttc ttctcggggt tggggtactt
     5461 gatgatgcgc tcgccgttgg tgtacacgat ccacttgctg cggatgccgc aggccttctg
     5521 ggtgaagctc ttgtagtcga agctgaactc gaacatgtcg tccttcttgt tgaagcggat
     5581 gaagtcgaag ccgtcgaaga actgcttggc cttctcgatg ttctcgtact tgatgtagaa
     5641 caggttcacg aagccggtgg tggggtcgat cttgctggtc aggtaggcgg gcacgtagta
     5701 gatgatgccg ctctgcttgc ccagctcggc gaagctcttg aacttgctgg tcagctgcag
     5761 ggcgttcagg gcgccgccca tcttctcggg gctcacctgc tcgcggctct tgtcgatcac
     5821 caggtagttc agcttctcga tcagcatctt ctcgaacttc tggtacacct gcttctccac
     5881 cttctggcgg ccgcgcttga agccgaagtt caggtcctcc aggcagatga tggcgttgta
     5941 cttcagcacc agcttggcca ccacgttcac cacctggctc aggtagccgg tcttcagctc
     6001 cttgatgttc tcgatggtgt tccagtcctt gcgggccttg ttgcggtcgt cctcgcgctc
     6061 gtccagcagg gcgtggtagt tggtctcgat gtcgtactcc ttgttgatga tgctgttcag
     6121 gctgatctgc tccaggatct tgccctcgct gttgatcacc accacgtaca gcaggttgcg
     6181 ctcgccgcgg tcgatgccga tcacgttcac gttgtcgtcg gtgcgcaggg cgttgttgat
     6241 cacgtcgttg aagcggcgca cctcgtccac gccgaagttc atggtgatgg ggatgtgcag
     6301 ggtgaacttg tacttgctgt agcgcttgtc cttcacgatg tcgtagctga aggtgctggt
     6361 ctccttcacc ttggcgcggt tggggttctt gttcttgatg ccctcgccgg ccttgtggat
     6421 caccagctcg ttctcggcga tgctggcggg gcggtagaac acctcggcct cgccgttcag
     6481 cttgtacacc acgttgtcca ggttgcgctg gtcgaacagc atcatgaagt acagggtgtg
     6541 caggttcagc ttgcccttgc tgtactcgct gaagtccttg ttgtagatct ggaacaggta
     6601 cagctcgttc ttctcgatca ggtcgttgat gtagctctcg tcgatgtcgg tgaaggtcag
     6661 cttgtagccc tgcttctcca cctcgcggta gaactcgctg atgtcgcggt agtcggcggt
     6721 gtcgctgaac tcgaagccga acttgctcca gtcctcgtgc ttcttgatgc tctccttgaa
     6781 gaagtcgatc aggttgtggc agtcgtcgat gctgaagctg gggcccttct tgtgggtgcc
     6841 cttcttgtag ttgctgtaca gctcggtgct ggggttgtag tagccgatgt tgctcttggc
     6901 gaagaacacc ttgggcagca tcttgttgct gccgggcagc agcttgtagt ccaccttctt
     6961 gaacacgttc tcggtcttgg cggcgggggg gttcacgaag gccttgttgg cggtggtgtt
     7021 catgatgccc aggtagtact tgccgtcctt gaagaacagg atgcccaggt tgtcggtctc
     7081 cttgttcttg tcccagccgt tcagcagggt gctcttgttg aagttcagct tcaccttctc
     7141 ggtgctgaag ggcttcttgg tcaggtagtt gcgggtcagg ttgtacaggc tgtccacggg
     7201 gcgcagctgc tccagggcct cctcctggcc cacgtacacc accaggttct tctccagctc
     7261 ctggccgctg ccgttgatca gcttgatgtc gtgctccagc tccttgatgc tgtccaggta
     7321 gtccttgatc accttcacgg ccttgatgtt cttgctcagc ttgcggctgc tgtcgtgctc
     7381 gttcaccacg atcttctcga actcgccgtt gtagatgcag atcttctcga tgtcctcgct
     7441 gatgcgctcg atgtagttct cgatggggct gatgtcctcc ttgctcaggt tgctcatctg
     7501 ctccagggtg tagctcttgt tcttcttcag ctccttctgg cgcttctcga agtacttgtc
     7561 gtccttcttc ttgttctcgt tggccaggtc gtactcctgg ttcagcagct cgtcgaaggc
     7621 gctccagctg ccgaacacga tgttgctcat ggtggtcttg ctcaggtcgt tcttcacgta
     7681 cacgttcttg ccctcgctct cgcgcagggc gtcgaagaag atgttgatct tctcgctctt
     7741 caggtaggtc atcagcacgt tgccgtaggc gccgatgctg ctcagcaggg tctcgtcgtc
     7801 cttgaactcg ccgatgaaca cctcctcgcg gtcgctcagg atctgcttgt acagcacctt
     7861 catcttgggg atcttcttga actcctccac cttgtggttc ttctggttgt acaggttgat
     7921 ggcgctgttc accttgccga tctggtagtt gtacatgtcg atgccctcct gggtcagggt
     7981 catgttgaag gtctccacca tgaacagggc gtcctgctcc tcctcctcga tgcagtcggc
     8041 cagcacgccg gcggccttca cgaaggcgta gctcttgatg ttgtccagga acttgggcag
     8101 gttctcgttg atcaggcggt aggccacggt gctgctcttc tcctcgtcgc tgtacaggtt
     8161 ctcgcgcacc ttgttgtagc tggtgaagta ggtgtagaag ttgcggaagc tctccagggc
     8221 gttgtagtcc tcctcgctga tgctgggcag cttctccagc aggtccagga tgtccttctt
     8281 gccgatcttg gtgtagttct cgtgctcctt caggcggccg gtcacctcgc ggcgcagcag
     8341 gtcctgggtc ttcttgaact cctcgcggtc ctccacgtcc acgtgcttct tcaggtagat
     8401 ctcggcggcc tggttcaggc tgggcagcat gtagttgtcc agggcctcgt tgatcagctg
     8461 cttgtggtac tcgtccatga tgcccttcac gtgctcgtag tcctgcttgc gcttcacgtc
     8521 gctctccagg atgttgttct tgcggatgtt ctccagggtc ttgccgatgg ggatcagctc
     8581 gttgcggatg gtcttgctca cggggtactt cttggtcagg ttctggtagt acatcccggg
     8641 aaccttcctc ttcttcttgg gcatattgga ttggaagtac aggttttcct cgatcccatt
     8701 agtctgcgcg tctttcaggg cttcatcgac agtctgacga ccgctggcgg cgttgatcac
     8761 cgcagtacgc acggcatacc agaaagcgga catctgcggg atgttcggca tgatttcacc
     8821 tttctgggcg ttttccatag tggcggcaat acgtggatct ttcgccaact cttcctcgta
     8881 agacttcagc gctacggcac ccagcggttt gtctttatta accgcttcca gaccttcatc
     8941 agtcagcaga tagttttcga ggaactcttt tgccagctct ttgttcggac tggcggcgtt
     9001 aatacctgcg ctcagcacgc caacgaacgg tttggatggt tgacccttga aggtcggcag
     9061 taccgttaca ccataattca ctttgctggt gtcgatgttg gaccatgccc acgggccgtt
     9121 gatggtcatc gctgtttcgc ctttattaaa ggcagcttct gcgatggagt aatcggtgtc
     9181 tgcattcatg tgtttgtttt taatcaggtc aaccaggaag gtcagacccg ctttcgcgcc
     9241 agcgttatcc acgcccacgt ctttaatgtc gtacttgccg ttttcatact tgaacgcata
     9301 acccccgtca gcagcaatca gcggccaggt gaagtacggt tcttgcaggt tgaacatcag
     9361 cgcgctctta cctttcgctt tcagttcttt atccagcgcc gggatctctt cccaggtttt
     9421 tggcgggttc ggcagcagat ctttgttata aatcagcgat aacgcttcaa cagcgatcgg
     9481 gtaagcaatc agcttgccgt tgtaacgtac ggcatcccag gtaaacggat acagcttgtc
     9541 ctggaacgct ttgtccgggg tgatttcagc caacaggcca gattgagcgt agccaccaaa
     9601 gcggtcgtgt gcccagaaga taatgtcagg gccatcgcca gttgccgcaa cctgtgggaa
     9661 tttctcttcc agtttatccg gatgctcaac ggtgacttta attccggtat ctttctcgaa
     9721 tttcttaccg acttcagcga gaccgttata gcctttatcg ccgttaatcc agattaccag
     9781 tttaccttct tcgattttca tagaagaacc atggtgatgg tgatggtgag aagaacccat
     9841 ggtatatctc cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc
     9901 acaattcccc tatagtgagt cgtattaatt tcgcgggatc gagatctcga tcctctacgc
     9961 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
    10021 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
    10081 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
    10141 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
    10201 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct
    10261 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
    10321 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
    10381 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
    10441 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
    10501 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
    10561 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
    10621 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
    10681 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
    10741 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
    10801 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
    10861 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
    10921 aatatctcac tcgcaatcaa attcagccga tagcggaacg gga
//
