LOCUS       sequence-373581-       12197 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses FLAG tagged SadCas9 KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12197)
  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 12197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: SadCas9 KRAB
FEATURES             Location/Qualifiers
     source          1..12197
                     /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             5603..5650
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5651..5674
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5708..5902
                     /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             5969..5989
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6002..6058
                     /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             6059..6655
                     /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     6552..6572
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6671..7259
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6724..6744)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7142..7153)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7331..7564
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7590..7607)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7596..7820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7866..8294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7953..7972)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8163..8184
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8303..8323)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8308..8637
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8488..8623
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8550..8569
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8685..8732
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8751..9125
                     /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    9255..9376
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9292..9311)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9346..9365
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9425..9441)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9425..9441)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9438..9460)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9449..9465
                     /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(9473..9479,9480..9497,9498..9503))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9518..9539
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9656..9673)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9827..10415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9907..9926)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10586..11377,11378..11446))
                     /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     11209..11228
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11447..11551)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11626..11645)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11817..12196
                     /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 accatgaagc ggaactacat
     2461 cctgggcctg gccatcggca tcaccagcgt gggctacggc atcatcgact acgagacacg
     2521 ggacgtgatc gatgccggcg tgcggctgtt caaagaggcc aacgtggaaa acaacgaggg
     2581 taggcggagc aagagaggcg ccagaaggct gaagcggcgg aggcggcata gaatccagag
     2641 agtgaagaag ctgctgttcg actacaacct gctgaccgac cacagcgagc tgagcggcat
     2701 caacccctac gaggccagag tgaagggcct gagccagaag ctgagcgagg aagagttctc
     2761 tgccgccctg ctgcacctgg ccaagagaag aggcgtgcac aacgtgaacg aggtggaaga
     2821 ggacaccggc aacgagctgt ccaccaaaga gcagatcagc cggaacagca aggccctgga
     2881 agagaaatac gtggccgaac tgcagctgga acggctgaag aaagacggcg aagtgcgggg
     2941 cagcatcaac agattcaaga ccagcgacta cgtgaaagaa gccaaacagc tgctgaaggt
     3001 gcagaaggcc taccaccagc tggaccagag cttcatcgac acctacatcg acctgctgga
     3061 aacccggcgg acctactatg agggacctgg cgagggcagc cccttcggct ggaaggacat
     3121 caaagaatgg tacgagatgc tgatgggcca ctgcacctac ttccccgagg aactgcggag
     3181 cgtgaagtac gcctacaacg ccgacctgta caacgccctg aacgacctga acaatctcgt
     3241 gatcaccagg gacgagaacg agaagctgga atattacgag aagttccaga tcatcgagaa
     3301 cgtgttcaag cagaagaaga agcccaccct gaagcagatc gccaaagaaa tcctcgtgaa
     3361 cgaagaggat attaagggct acagagtgac cagcaccggc aagcccgagt tcaccaacct
     3421 gaaggtgtac cacgacatca aggacattac cgcccggaaa gagattattg agaacgccga
     3481 gctgctggat cagattgcca agatcctgac catctaccag agcagcgagg acatccagga
     3541 agaactgacc aatctgaact ccgagctgac ccaggaagag atcgagcaga tctctaatct
     3601 gaagggctat accggcaccc acaacctgag cctgaaggcc atcaacctga tcctggacga
     3661 gctgtggcac accaacgaca accagatcgc tatcttcaac cggctgaagc tggtgcccaa
     3721 gaaggtggac ctgtcccagc agaaagagat ccccaccacc ctggtggacg acttcatcct
     3781 gagccccgtc gtgaagagaa gcttcatcca gagcatcaaa gtgatcaacg ccatcatcaa
     3841 gaagtacggc ctgcccaacg acatcattat cgagctggcc cgcgagaaga actccaagga
     3901 cgcccagaaa atgatcaacg agatgcagaa gcggaaccgg cagaccaacg agcggatcga
     3961 ggaaatcatc cggaccaccg gcaaagagaa cgccaagtac ctgatcgaga agatcaagct
     4021 gcacgacatg caggaaggca agtgcctgta cagcctggaa gccatccctc tggaagatct
     4081 gctgaacaac cccttcaact atgaggtgga ccacatcatc cccagaagcg tgtccttcga
     4141 caacagcttc aacaacaagg tgctcgtgaa gcaggaagaa gccagcaaga agggcaaccg
     4201 gaccccattc cagtacctga gcagcagcga cagcaagatc agctacgaaa ccttcaagaa
     4261 gcacatcctg aatctggcca agggcaaggg cagaatcagc aagaccaaga aagagtatct
     4321 gctggaagaa cgggacatca acaggttctc cgtgcagaaa gacttcatca accggaacct
     4381 ggtggatacc agatacgcca ccagaggcct gatgaacctg ctgcggagct acttcagagt
     4441 gaacaacctg gacgtgaaag tgaagtccat caatggcggc ttcaccagct ttctgcggcg
     4501 gaagtggaag tttaagaaag agcggaacaa ggggtacaag caccacgccg aggacgccct
     4561 gatcattgcc aacgccgatt tcatcttcaa agagtggaag aaactggaca aggccaaaaa
     4621 agtgatggaa aaccagatgt tcgaggaaaa gcaggccgag agcatgcccg agatcgaaac
     4681 cgagcaggag tacaaagaga tcttcatcac cccccaccag atcaagcaca ttaaggactt
     4741 caaggactac aagtacagcc accgggtgga caagaagcct aatagagagc tgattaacga
     4801 caccctgtac tccacccgga aggacgacaa gggcaacacc ctgatcgtga acaatctgaa
     4861 cggcctgtac gacaaggaca atgacaagct gaaaaagctg atcaacaaga gccccgaaaa
     4921 gctgctgatg taccaccacg acccccagac ctaccagaaa ctgaagctga ttatggaaca
     4981 gtacggcgac gagaagaatc ccctgtacaa gtactacgag gaaaccggga actacctgac
     5041 caagtactcc aaaaaggaca acggccccgt gatcaagaag attaagtatt acggcaacaa
     5101 actgaacgcc catctggaca tcaccgacga ctaccccaac agcagaaaca aggtcgtgaa
     5161 gctgtccctg aagccctaca gattcgacgt gtacctggac aatggcgtgt acaagttcgt
     5221 gaccgtgaag aatctggatg tgatcaaaaa agaaaactac tacgaagtga atagcaagtg
     5281 ctatgaggaa gctaagaagc tgaagaagat cagcaaccag gccgagttta tcgcctcctt
     5341 ctacaacaac gatctgatca agatcaacgg cgagctgtat agagtgatcg gcgtgaacaa
     5401 cgacctgctg aaccggatcg aagtgaacat gatcgacatc acctaccgcg agtacctgga
     5461 aaacatgaac gacaagaggc cccccaggat cattaagaca atcgcctcca agacccagag
     5521 cattaagaag tacagcacag acattctggg caacctgtat gaagtgaaat ctaagaagca
     5581 ccctcagatc atcaaaaagg gcaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa
     5641 aaagaaaaag gattacaaag acgatgacga taagggatcc gatgctaagt cactgactgc
     5701 ctggtcccgg acactggtga ccttcaagga tgtgtttgtg gacttcacca gggaggagtg
     5761 gaagctgctg gacactgctc agcagatcct gtacagaaat gtgatgctgg agaactataa
     5821 gaacctggtt tccttgggtt atcagcttac taagccagat gtgatcctcc ggttggagaa
     5881 gggagaagag ccctggctgg tggagagaga aattcaccaa gagacccatc ctgattcaga
     5941 gactgcattt gaaatcaaat catcagttcc gaaaaagaaa cgcaaagttg ctagctccgg
     6001 cgcaacaaac ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgac
     6061 cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac
     6121 cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca
     6181 catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg
     6241 caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt
     6301 cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg
     6361 gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc
     6421 gtggttcctg gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc
     6481 cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac
     6541 ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt
     6601 cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgaacgcg
     6661 ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     6721 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     6781 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     6841 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     6901 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     6961 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     7021 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     7081 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     7141 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     7201 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     7261 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     7321 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     7381 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     7441 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     7501 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     7561 agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     7621 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     7681 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     7741 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     7801 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     7861 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     7921 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     7981 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     8041 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     8101 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     8161 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     8221 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     8281 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     8341 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     8401 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     8461 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     8521 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     8581 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     8641 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
     8701 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt
     8761 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga
     8821 ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg
     8881 acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg
     8941 cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca
     9001 cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc
     9061 gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg
     9121 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     9181 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     9241 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     9301 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     9361 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     9421 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     9481 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     9541 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     9601 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     9661 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     9721 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     9781 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     9841 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     9901 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     9961 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
    10021 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
    10081 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
    10141 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
    10201 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
    10261 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
    10321 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
    10381 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
    10441 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
    10501 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
    10561 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
    10621 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
    10681 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
    10741 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
    10801 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
    10861 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
    10921 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
    10981 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
    11041 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
    11101 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
    11161 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
    11221 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
    11281 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
    11341 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    11401 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
    11461 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
    11521 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
    11581 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
    11641 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
    11701 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
    11761 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
    11821 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
    11881 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
    11941 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
    12001 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
    12061 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
    12121 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
    12181 catcgctatt accatgg
//
