LOCUS       sequence-298975-       11137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression plasmid of AsCas12a-6xNLS-E174R/S542R with 
            puromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11137)
  AUTHORS   Gier RA, Budinich KA, Evitt NH, Cao Z, Freilich ES, Chen Q, Qi J, 
            Lan Y, Kohli RM, Shi J
  TITLE     High-performance CRISPR-Cas12a genome editing for combinatorial 
            genetic screening.
  JOURNAL   Nat Commun. 2020 Jul 13;11(1):3455. doi: 10.1038/s41467-020-17209-1.
  PUBMED    32661245
REFERENCE   2  (bases 1 to 11137)
  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: "Nat 
            Commun."; date: "2020-07-13"; volume: "11(1):3455. doi"; pages: " 
            10.1038/s41467-020-17209-1"
COMMENT     Sequence Label: AsCas12a-6xNLS-E174R/S542R (pRG232)
FEATURES             Location/Qualifiers
     source          1..11137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     240..260
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        288..514
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             515..695
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    742..867
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1360..1593
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1778..1822
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2006..2123
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2192..2403
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2423..2432
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2432..2455
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2456..6373
                     /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
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      6093..6102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6383..6409
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6419..6445
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6455..6481
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6491..6517
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6527..6553
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6563..6589
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6605..6661
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6662..7261
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6662..6681)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7158..7178
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7277..7865
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7330..7350)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7748..7759)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7937..8170
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8248..8369
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8285..8304)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8339..8358
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8409..8544
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8564..8583)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8565..8583)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8593..8610)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8593..8609)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8602..8624)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8751..9206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8838..8857)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9048..9069
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9232..9336
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9337..9405,9406..10197)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9555..9574)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10368..10956
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10857..10876
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11110..11127
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
      241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
      301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
      361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
      421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
      481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
      541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1981 aaccccgagg ggacccagac cacaatttta aaagaaaagg ggggattggg gggtacagtg
     2041 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     2101 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc
     2161 gccggctcga gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     2221 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     2281 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     2341 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     2401 caggtgtcgt gacgcgggat ccgccaccat ggattacaaa gacgatgacg ataagacaca
     2461 gttcgagggc tttaccaacc tgtatcaggt gagcaagaca ctgcggtttg agctgatccc
     2521 acagggcaag accctgaagc acatccagga gcagggcttc atcgaggagg acaaggcccg
     2581 caatgatcac tacaaggagc tgaagcccat catcgatcgg atctacaaga cctatgccga
     2641 ccagtgcctg cagctggtgc agctggattg ggagaacctg agcgccgcca tcgactccta
     2701 tagaaaggag aaaaccgagg agacaaggaa cgccctgatc gaggagcagg ccacatatcg
     2761 caatgccatc cacgactact tcatcggccg gacagacaac ctgaccgatg ccatcaataa
     2821 gagacacgcc gagatctaca agggcctgtt caaggccgag ctgtttaatg gcaaggtgct
     2881 gaagcagctg ggcaccgtga ccacaaccga gcacgagaac gccctgctgc ggagcttcga
     2941 caagtttaca acctacttct ccggctttta tcggaacagg aagaacgtgt tcagcgccga
     3001 ggatatcagc acagccatcc cacaccgcat cgtgcaggac aacttcccca agtttaagga
     3061 gaattgtcac atcttcacac gcctgatcac cgccgtgccc agcctgcggg agcactttga
     3121 gaacgtgaag aaggccatcg gcatcttcgt gagcacctcc atcgaggagg tgttttcctt
     3181 ccctttttat aaccagctgc tgacacagac ccagatcgac ctgtataacc agctgctggg
     3241 aggaatctct cgggaggcag gcaccgagaa gatcaagggc ctgaacgagg tgctgaatct
     3301 ggccatccag aagaatgatg agacagccca catcatcgcc tccctgccac acagattcat
     3361 ccccctgttt aagcagatcc tgtccgatag gaacaccctg tctttcatcc tggaggagtt
     3421 taagagcgac gaggaagtga tccagtcctt ctgcaagtac aagacactgc tgagaaacga
     3481 gaacgtgctg gagacagccg aggccctgtt taacgagctg aacagcatcg acctgacaca
     3541 catcttcatc agccacaaga agctggagac aatcagcagc gccctgtgcg accactggga
     3601 tacactgagg aatgccctgt atgagcggag aatctccgag ctgacaggca agatcaccaa
     3661 gtctgccaag gagaaggtgc agcgcagcct gaagcacgag gatatcaacc tgcaggagat
     3721 catctctgcc gcaggcaagg agctgagcga ggccttcaag cagaaaacca gcgagatcct
     3781 gtcccacgca cacgccgccc tggatcagcc actgcctaca accctgaaga agcaggagga
     3841 gaaggagatc ctgaagtctc agctggacag cctgctgggc ctgtaccacc tgctggactg
     3901 gtttgccgtg gatgagtcca acgaggtgga ccccgagttc tctgcccggc tgaccggcat
     3961 caagctggag atggagcctt ctctgagctt ctacaacaag gccagaaatt atgccaccaa
     4021 gaagccctac tccgtggaga agttcaagct gaactttcag atgcctacac tggcccgtgg
     4081 ctgggacgtg aataaggaga agaacaatgg cgccatcctg tttgtgaaga acggcctgta
     4141 ctatctgggc atcatgccaa agcagaaggg caggtataag gccctgagct tcgagcccac
     4201 agagaaaacc agcgagggct ttgataagat gtactatgac tacttccctg atgccgccaa
     4261 gatgatccca aagtgcagca cccagctgaa ggccgtgaca gcccactttc agacccacac
     4321 aacccccatc ctgctgtcca acaatttcat cgagcctctg gagatcacaa aggagatcta
     4381 cgacctgaac aatcctgaga aggagccaaa gaagtttcag acagcctacg ccaagaaaac
     4441 cggcgaccag aagggctaca gagaggccct gtgcaagtgg atcgacttca caagggattt
     4501 tctgtccaag tataccaaga caacctctat cgatctgtct agcctgcggc catcctctca
     4561 gtataaggac ctgggcgagt actatgccga gctgaatccc ctgctgtacc acatcagctt
     4621 ccagagaatc gccgagaagg agatcatgga tgccgtggag acaggcaagc tgtacctgtt
     4681 ccagatctat aacaaggact ttgccaaggg ccaccacggc aagcctaatc tgcacacact
     4741 gtattggacc ggcctgtttt ctccagagaa cctggccaag acaagcatca agctgaatgg
     4801 ccaggccgag ctgttctacc gccctaagtc caggatgaag aggatggcac accggctggg
     4861 agagaagatg ctgaacaaga agctgaagga tcagaaaacc ccaatccccg acaccctgta
     4921 ccaggagctg tacgactatg tgaatcacag actgtcccac gacctgtctg atgaggccag
     4981 ggccctgctg cccaacgtga tcaccaagga ggtgtctcac gagatcatca aggataggcg
     5041 ctttaccagc gacaagttct ttttccacgt gcctatcaca ctgaactatc aggccgccaa
     5101 ttccccatct aagttcaacc agagggtgaa tgcctacctg aaggagcacc ccgagacacc
     5161 tatcatcggc atcgatcggg gcgagagaaa cctgatctat atcacagtga tcgactccac
     5221 cggcaagatc ctggagcagc ggagcctgaa caccatccag cagtttgatt accagaagaa
     5281 gctggacaac agggagaagg agagggtggc agcaaggcag gcctggtctg tggtgggcac
     5341 aatcaaggat ctgaagcagg gctatctgag ccaggtcatc cacgagatcg tggacctgat
     5401 gatccactac caggccgtgg tggtgctgga gaacctgaat ttcggcttta agagcaagag
     5461 gaccggcatc gccgagaagg ccgtgtacca gcagttcgag aagatgctga tcgataagct
     5521 gaattgcctg gtgctgaagg actatccagc agagaaagtg ggaggcgtgc tgaacccata
     5581 ccagctgaca gaccagttca cctcctttgc caagatgggc acccagtctg gcttcctgtt
     5641 ttacgtgcct gccccatata catctaagat cgatcccctg accggcttcg tggacccctt
     5701 cgtgtggaaa accatcaaga atcacgagag ccgcaagcac ttcctggagg gcttcgactt
     5761 tctgcactac gacgtgaaaa ccggcgactt catcctgcac tttaagatga acagaaatct
     5821 gtccttccag aggggcctgc ccggctttat gcctgcatgg gatatcgtgt tcgagaagaa
     5881 cgagacacag tttgacgcca agggcacccc tttcatcgcc ggcaagagaa tcgtgccagt
     5941 gatcgagaat cacagattca ccggcagata ccgggacctg tatcctgcca acgagctgat
     6001 cgccctgctg gaggagaagg gcatcgtgtt cagggatggc tccaacatcc tgccaaagct
     6061 gctggagaat gacgattctc acgccatcga caccatggtg gccctgatcc gcagcgtgct
     6121 gcagatgcgg aactccaatg ccgccacagg cgaggactat atcaacagcc ccgtgcgcga
     6181 tctgaatggc gtgtgcttcg actcccggtt tcagaaccca gagtggccca tggacgccga
     6241 tgccaatggc gcctaccaca tcgccctgaa gggccagctg ctgctgaatc acctgaagga
     6301 gagcaaggat ctgaagctgc agaacggcat ctccaatcag gactggctgg cctacatcca
     6361 ggagctgcgc aacggcggct cccctgctgc taaacgtgtt aagcttgatg ggggtagccc
     6421 ggcagccaag agagtcaaac tcgacggcgg ctccccagcg gcaaaaaggg tgaaactaga
     6481 cgggggtagc cccgccgcga agcgtgtaaa gctggatgga ggatcgcctg cggctaagcg
     6541 agtcaaatta gacggtgggt caccggctgc aaagcgcgtc aaattggatg ctagcggcag
     6601 cggcgccacc aacttcagcc tgctgaagca ggccggcgac gtggaggaga accccggccc
     6661 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     6721 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     6781 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     6841 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     6901 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     6961 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     7021 gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg
     7081 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     7141 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     7201 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     7261 aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     7321 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     7381 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     7441 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     7501 tgacgcaacc cccactggtt gggacattgc caccacctgt cagctccttt ccgggacttt
     7561 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     7621 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     7681 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     7741 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     7801 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     7861 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     7921 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     7981 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     8041 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     8101 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     8161 atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca
     8221 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     8281 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     8341 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     8401 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     8461 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     8521 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     8581 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     8641 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     8701 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     8761 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     8821 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     8881 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     8941 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     9001 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     9061 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     9121 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     9181 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     9241 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     9301 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     9361 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     9421 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     9481 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     9541 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     9601 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     9661 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     9721 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     9781 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     9841 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     9901 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     9961 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
    10021 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
    10081 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
    10141 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
    10201 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
    10261 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
    10321 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
    10381 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
    10441 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
    10501 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
    10561 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
    10621 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
    10681 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
    10741 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
    10801 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
    10861 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
    10921 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
    10981 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
    11041 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
    11101 accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
