LOCUS       sequence-373525-       12959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG tagged AsdCas12a KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12959)
  AUTHORS   Escobar M, Li J, Patel A, Liu S, Xu Q, Hilton IB
  TITLE     Quantification of Genome Editing and Transcriptional Control 
            Capabilities Reveals Hierarchies among Diverse CRISPR/Cas Systems in
            Human Cells.
  JOURNAL   ACS Synth Biol. 2022 Oct 21;11(10):3239-3250. doi: 
            10.1021/acssynbio.2c00156. Epub 2022 Sep 26.
  PUBMED    36162812
REFERENCE   2  (bases 1 to 12959)
  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: 
            "10.1021/acssynbio.2c00156"; journalName: "ACS Synth Biol"; date: 
            "2022-10-21- 21"; volume: "11"; issue: "10"; pages: "3239-3250"
COMMENT     Sequence Label: AsdCas12a KRAB
FEATURES             Location/Qualifiers
     source          1..12959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             217..397
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    444..569
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1062..1295
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1480..1524
                     /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    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        2071..2146
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2208..2419
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2444..6364
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIARGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      6084..6093
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6365..6412
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6413..6436
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6470..6664
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             6731..6751
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6764..6820
                     /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             6821..7417
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     7314..7334
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7433..8021
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7486..7506)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7904..7915)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8093..8326
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8352..8369)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8358..8582
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8628..9056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8715..8734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8925..8946
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9065..9085)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9070..9399
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9250..9385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9312..9331
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9447..9494
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9513..9887
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10017..10138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10054..10073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10108..10127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10187..10203)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10187..10203)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10200..10222)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10211..10227
                     /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)."
     promoter        complement(join(10235..10241,10242..10259,10260..10265))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10280..10301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10418..10435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10589..11177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10669..10688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11348..12139,12140..12208))
                     /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     11971..11990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12209..12313)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12388..12407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12579..12958
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      241 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      301 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      361 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      421 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      481 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
      541 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
      601 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
      661 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
      721 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
      781 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
      841 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
      901 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
      961 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1021 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1081 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1141 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1201 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1261 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1321 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1381 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1441 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1501 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1561 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1621 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1681 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1741 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     1801 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     1861 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     1921 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     1981 taaggtaccg atatcactag tgttccatta actgcgctaa aaataatttt taaatctttt
     2041 ttagcttctt gctctttttt gtacgtctct gttttagagc tagaaatagc aagttaaaat
     2101 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcgc
     2161 tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     2221 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag
     2281 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     2341 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     2401 gtttgccgcc agaacacagg accggttcta gagcgctgcc accatgacac agttcgaggg
     2461 ctttaccaac ctgtatcagg tgagcaagac actgcggttt gagctgatcc cacagggcaa
     2521 gaccctgaag cacatccagg agcagggctt catcgaggag gacaaggccc gcaatgatca
     2581 ctacaaggag ctgaagccca tcatcgatcg gatctacaag acctatgccg accagtgcct
     2641 gcagctggtg cagctggatt gggagaacct gagcgccgcc atcgactcct atagaaagga
     2701 gaaaaccgag gagacaagga acgccctgat cgaggagcag gccacatatc gcaatgccat
     2761 ccacgactac ttcatcggcc ggacagacaa cctgaccgat gccatcaata agagacacgc
     2821 cgagatctac aagggcctgt tcaaggccga gctgtttaat ggcaaggtgc tgaagcagct
     2881 gggcaccgtg accacaaccg agcacgagaa cgccctgctg cggagcttcg acaagtttac
     2941 aacctacttc tccggctttt atgagaacag gaagaacgtg ttcagcgccg aggatatcag
     3001 cacagccatc ccacaccgca tcgtgcagga caacttcccc aagtttaagg agaattgtca
     3061 catcttcaca cgcctgatca ccgccgtgcc cagcctgcgg gagcactttg agaacgtgaa
     3121 gaaggccatc ggcatcttcg tgagcacctc catcgaggag gtgttttcct tcccttttta
     3181 taaccagctg ctgacacaga cccagatcga cctgtataac cagctgctgg gaggaatctc
     3241 tcgggaggca ggcaccgaga agatcaaggg cctgaacgag gtgctgaatc tggccatcca
     3301 gaagaatgat gagacagccc acatcatcgc ctccctgcca cacagattca tccccctgtt
     3361 taagcagatc ctgtccgata ggaacaccct gtctttcatc ctggaggagt ttaagagcga
     3421 cgaggaagtg atccagtcct tctgcaagta caagacactg ctgagaaacg agaacgtgct
     3481 ggagacagcc gaggccctgt ttaacgagct gaacagcatc gacctgacac acatcttcat
     3541 cagccacaag aagctggaga caatcagcag cgccctgtgc gaccactggg atacactgag
     3601 gaatgccctg tatgagcgga gaatctccga gctgacaggc aagatcacca agtctgccaa
     3661 ggagaaggtg cagcgcagcc tgaagcacga ggatatcaac ctgcaggaga tcatctctgc
     3721 cgcaggcaag gagctgagcg aggccttcaa gcagaaaacc agcgagatcc tgtcccacgc
     3781 acacgccgcc ctggatcagc cactgcctac aaccctgaag aagcaggagg agaaggagat
     3841 cctgaagtct cagctggaca gcctgctggg cctgtaccac ctgctggact ggtttgccgt
     3901 ggatgagtcc aacgaggtgg accccgagtt ctctgcccgg ctgaccggca tcaagctgga
     3961 gatggagcct tctctgagct tctacaacaa ggccagaaat tatgccacca agaagcccta
     4021 ctccgtggag aagttcaagc tgaactttca gatgcctaca ctggcctctg gctgggacgt
     4081 gaataaggag aagaacaatg gcgccatcct gtttgtgaag aacggcctgt actatctggg
     4141 catcatgcca aagcagaagg gcaggtataa ggccctgagc ttcgagccca cagagaaaac
     4201 cagcgagggc tttgataaga tgtactatga ctacttccct gatgccgcca agatgatccc
     4261 aaagtgcagc acccagctga aggccgtgac agcccacttt cagacccaca caacccccat
     4321 cctgctgtcc aacaatttca tcgagcctct ggagatcaca aaggagatct acgacctgaa
     4381 caatcctgag aaggagccaa agaagtttca gacagcctac gccaagaaaa ccggcgacca
     4441 gaagggctac agagaggccc tgtgcaagtg gatcgacttc acaagggatt ttctgtccaa
     4501 gtataccaag acaacctcta tcgatctgtc tagcctgcgg ccatcctctc agtataagga
     4561 cctgggcgag tactatgccg agctgaatcc cctgctgtac cacatcagct tccagagaat
     4621 cgccgagaag gagatcatgg atgccgtgga gacaggcaag ctgtacctgt tccagatcta
     4681 taacaaggac tttgccaagg gccaccacgg caagcctaat ctgcacacac tgtattggac
     4741 cggcctgttt tctccagaga acctggccaa gacaagcatc aagctgaatg gccaggccga
     4801 gctgttctac cgccctaagt ccaggatgaa gaggatggca caccggctgg gagagaagat
     4861 gctgaacaag aagctgaagg atcagaaaac cccaatcccc gacaccctgt accaggagct
     4921 gtacgactat gtgaatcaca gactgtccca cgacctgtct gatgaggcca gggccctgct
     4981 gcccaacgtg atcaccaagg aggtgtctca cgagatcatc aaggataggc gctttaccag
     5041 cgacaagttc tttttccacg tgcctatcac actgaactat caggccgcca attccccatc
     5101 taagttcaac cagagggtga atgcctacct gaaggagcac cccgagacac ctatcatcgg
     5161 catcgcccgg ggcgagagaa acctgatcta tatcacagtg atcgactcca ccggcaagat
     5221 cctggagcag cggagcctga acaccatcca gcagtttgat taccagaaga agctggacaa
     5281 cagggagaag gagagggtgg cagcaaggca ggcctggtct gtggtgggca caatcaagga
     5341 tctgaagcag ggctatctga gccaggtcat ccacgagatc gtggacctga tgatccacta
     5401 ccaggccgtg gtggtgctgg agaacctgaa tttcggcttt aagagcaaga ggaccggcat
     5461 cgccgagaag gccgtgtacc agcagttcga gaagatgctg atcgataagc tgaattgcct
     5521 ggtgctgaag gactatccag cagagaaagt gggaggcgtg ctgaacccat accagctgac
     5581 agaccagttc acctcctttg ccaagatggg cacccagtct ggcttcctgt tttacgtgcc
     5641 tgccccatat acatctaaga tcgatcccct gaccggcttc gtggacccct tcgtgtggaa
     5701 aaccatcaag aatcacgaga gccgcaagca cttcctggag ggcttcgact ttctgcacta
     5761 cgacgtgaaa accggcgact tcatcctgca ctttaagatg aacagaaatc tgtccttcca
     5821 gaggggcctg cccggcttta tgcctgcatg ggatatcgtg ttcgagaaga acgagacaca
     5881 gtttgacgcc aagggcaccc ctttcatcgc cggcaagaga atcgtgccag tgatcgagaa
     5941 tcacagattc accggcagat accgggacct gtatcctgcc aacgagctga tcgccctgct
     6001 ggaggagaag ggcatcgtgt tcagggatgg ctccaacatc ctgccaaagc tgctggagaa
     6061 tgacgattct cacgccatcg acaccatggt ggccctgatc cgcagcgtgc tgcagatgcg
     6121 gaactccaat gccgccacag gcgaggacta tatcaacagc cccgtgcgcg atctgaatgg
     6181 cgtgtgcttc gactcccggt ttcagaaccc agagtggccc atggacgccg atgccaatgg
     6241 cgcctaccac atcgccctga agggccagct gctgctgaat cacctgaagg agagcaagga
     6301 tctgaagctg cagaacggca tctccaatca ggactggctg gcctacatcc aggagctgcg
     6361 caacaagcga cctgccgcca caaagaaggc tggacaggct aagaagaaga aagattacaa
     6421 agacgatgac gataagggat ccgatgctaa gtcactgact gcctggtccc ggacactggt
     6481 gaccttcaag gatgtgtttg tggacttcac cagggaggag tggaagctgc tggacactgc
     6541 tcagcagatc ctgtacagaa atgtgatgct ggagaactat aagaacctgg tttccttggg
     6601 ttatcagctt actaagccag atgtgatcct ccggttggag aagggagaag agccctggct
     6661 ggtggagaga gaaattcacc aagagaccca tcctgattca gagactgcat ttgaaatcaa
     6721 atcatcagtt ccgaaaaaga aacgcaaagt tgctagctcc ggcgcaacaa acttctctct
     6781 gctgaaacaa gccggagatg tcgaagagaa tcctggaccg accgagtaca agcccacggt
     6841 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     6901 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     6961 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     7021 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     7081 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     7141 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     7201 cggagtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     7261 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     7321 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     7381 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc
     7441 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     7501 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     7561 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     7621 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     7681 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     7741 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     7801 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     7861 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     7921 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     7981 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg
     8041 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     8101 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
     8161 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     8221 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     8281 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac
     8341 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     8401 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     8461 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     8521 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     8581 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     8641 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     8701 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     8761 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     8821 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     8881 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     8941 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     9001 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     9061 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     9121 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     9181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     9241 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     9301 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     9361 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     9421 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     9481 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     9541 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     9601 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     9661 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     9721 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     9781 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     9841 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     9901 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     9961 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
    10021 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
    10081 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
    10141 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
    10201 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
    10261 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
    10321 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
    10381 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
    10441 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
    10501 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
    10561 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
    10621 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
    10681 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
    10741 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
    10801 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
    10861 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
    10921 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
    10981 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
    11041 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
    11101 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
    11161 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
    11221 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
    11281 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
    11341 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
    11401 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
    11461 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
    11521 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
    11581 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
    11641 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
    11701 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
    11761 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
    11821 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
    11881 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
    11941 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
    12001 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
    12061 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
    12121 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
    12181 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
    12241 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
    12301 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
    12361 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
    12421 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
    12481 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
    12541 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
    12601 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
    12661 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
    12721 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
    12781 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
    12841 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
    12901 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgg
//
