LOCUS       Exported               14914 bp ds-DNA     circular SYN 25-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14914)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Oct 3, 2023 from SnapGene 4.3.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14914
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..199
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     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."
     misc_feature    1822..1939
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     1851..1871
                     /label=#421-cPPT/CTS-F
     promoter        2039..2250
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     2236..2269
                     /label=XbaI-AfeI-F
     primer_bind     2276..2295
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2276..2294
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2308..2343
                     /codon_start=1
                     /label=Feature 16
                     /translation="MKRTADGSEFES"
     CDS             2344..2364
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2365..6465
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(2392..2410)
                     /label=#423-Cas9-R
     primer_bind     3329..3348
                     /label=oAT21 pCMV-PE2 1686-1705 F2
     primer_bind     complement(3532..3552)
                     /label=oAT20 pCMV-PE2 1889-1909 R1
     primer_bind     complement(3567..3634)
                     /label=#44-WPRE-R1
     primer_bind     4390..4413
                     /label=oAT22 pCMV-PE2 2747-2770 F3
     primer_bind     complement(4738..4758)
                     /label=oAT23 pCMV-PE2 3095-3115 R2
     primer_bind     5472..5496
                     /label=oAT24 pCMV-PE1 3829-3853 F4
     primer_bind     complement(5737..5756)
                     /label=oAT25 pCMV-PE2 4094-4113 R3
     primer_bind     5892..5912
                     /label=#228-Cas9-F
     primer_bind     6400..6419
                     /label=oAT26 pCMV-PE1 4757-4776 F5
     CDS             6565..8637
                     /codon_start=1
                     /label=M-MLV
                     /translation="TLNIEDEYRLHETSKEPDVSLGSTWLSDFPQAWAETGGMGLAVRQ
                     APLIIPLKATSTPVSIKQYPMSQEARLGIKPHIQRLLDQGILVPCQSPWNTPLLPVKKP
                     GTNDYRPVQDLREVNKRVEDIHPTVPNPYNLLSGLPPSHQWYTVLDLKDAFFCLRLHPT
                     SQPLFAFEWRDPEMGISGQLTWTRLPQGFKNSPTLFNEALHRDLADFRIQHPDLILLQY
                     VDDLLLAATSELDCQQGTRALLQTLGNLGYRASAKKAQICQKQVKYLGYLLKEGQRWLT
                     EARKETVMGQPTPKTPRQLREFLGKAGFCRLFIPGFAEMAAPLYPLTKPGTLFNWGPDQ
                     QKAYQEIKQALLTAPALGLPDLTKPFELFVDEKQGYAKGVLTQKLGPWRRPVAYLSKKL
                     DPVAAGWPPCLRMVAAIAVLTKDAGKLTMGQPLVILAPHAVEALVKQPPDRWLSNARMT
                     HYQALLLDTDRVQFGPVVALNPATLLPLPEEGLQHNCLDILAEAHGTRPDLTDQPLPDA
                     DHTWYTDGSSLLQEGQRKAGAAVTTETEVIWAKALPAGTSAQRAELIALTQALKMAEGK
                     KLNVYTDSRYAFATAHIHGEIYRRRGWLTSEGKEIKNKDEILALLKALFLPKRLSIIHC
                     PGHQKGHSAEARGNRMADQAARKAAITETPDTSTLLIENSSPSGGSKRTADGSEFE"
     primer_bind     complement(6636..6657)
                     /label=oAT27 pCMV-PE1 4993-5014 REV 4
     exon            7162..7164
                     /label=D200N
     primer_bind     7413..7434
                     /label=oAT28 pCMV-PE1 5770-5791 F6
     exon            7480..7482
                     /label=T306K
     exon            7501..7503
                     /label=W313F
     exon            7552..7554
                     /label=T330P
     primer_bind     complement(7793..7812)
                     /label=oAT29 pCMV-PE1 6150-6169 REV 5
     exon            8371..8373
                     /label=L603W
     primer_bind     8612..8658
                     /label=#263-Flag-Puro-F
     misc_feature    8629..8633
                     /label=EcoRI
     CDS             8638..8685
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             8686..8709
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8719..8775
                     /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             8776..9372
                     /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     complement(8815..8834)
                     /label=Flag-Puro-R
     misc_feature    9388..9976
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(9859..9870)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9874..9913)
                     /label=#44-WPRE-R1
     LTR             10048..10281
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10313..10537
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(10387..10411)
                     /label=oAT30 pCMV-PE1 7144-7168 R6
     rep_origin      10583..11011
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        11025..11354
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11205..11340
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        11402..11449
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11468..11842
                     /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    11972..12093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12142..12158)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    12166..12182
                     /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(12190..12220)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12235..12256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(12544..13132)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(13303..14163)
                     /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"
     promoter        complement(14164..14268)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        14534..14913
                     /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 taagaattcg ctagctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     2041 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc
     2101 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     2161 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     2221 tttcgcaacg ggtttgccgc cagaacacag gaccggttct agaagcgctg gccgctaata
     2281 cgactcacta tagggagagc cgccaccatg aaacggacag ccgacggaag cgagttcgag
     2341 tcaccaaaga agaagcggaa agtcgacaag aagtacagca tcggcctgga catcggcacc
     2401 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     2461 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     2521 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     2581 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     2641 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     2701 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     2761 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     2821 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     2881 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     2941 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     3001 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     3061 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     3121 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     3181 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     3241 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     3301 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     3361 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     3421 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     3481 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     3541 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     3601 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     3661 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     3721 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     3781 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     3841 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     3901 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     3961 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     4021 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     4081 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     4141 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     4201 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     4261 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     4321 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     4381 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     4441 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     4501 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     4561 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     4621 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     4681 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     4741 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     4801 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     4861 tccgactacg atgtggacgc tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     4921 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     4981 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     5041 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     5101 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     5161 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     5221 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     5281 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     5341 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     5401 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     5461 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     5521 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     5581 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     5641 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     5701 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     5761 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     5821 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     5881 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     5941 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     6001 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     6061 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     6121 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     6181 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     6241 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     6301 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     6361 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     6421 ggcctgtacg agacacggat cgacctgtct cagctgggag gtgactctgg aggatctagc
     6481 ggaggatcct ctggcagcga gacaccagga acaagcgagt cagcaacacc agagagcagt
     6541 ggcggcagca gcggcggcag cagcacccta aatatagaag atgagtatcg gctacatgag
     6601 acctcaaaag agccagatgt ttctctaggg tccacatggc tgtctgattt tcctcaggcc
     6661 tgggcggaaa ccgggggcat gggactggca gttcgccaag ctcctctgat catacctctg
     6721 aaagcaacct ctacccccgt gtccataaaa caatacccca tgtcacaaga agccagactg
     6781 gggatcaagc cccacataca gagactgttg gaccagggaa tactggtacc ctgccagtcc
     6841 ccctggaaca cgcccctgct acccgttaag aaaccaggga ctaatgatta taggcctgtc
     6901 caggatctga gagaagtcaa caagcgggtg gaagacatcc accccaccgt gcccaaccct
     6961 tacaacctct tgagcgggct cccaccgtcc caccagtggt acactgtgct tgatttaaag
     7021 gatgcctttt tctgcctgag actccacccc accagtcagc ctctcttcgc ctttgagtgg
     7081 agagatccag agatgggaat ctcaggacaa ttgacctgga ccagactccc acagggtttc
     7141 aaaaacagtc ccaccctgtt taatgaggca ctgcacagag acctagcaga cttccggatc
     7201 cagcacccag acttgatcct gctacagtac gtggatgact tactgctggc cgccacttct
     7261 gagctagact gccaacaagg tactcgggcc ctgttacaaa ccctagggaa cctcgggtat
     7321 cgggcctcgg ccaagaaagc ccaaatttgc cagaaacagg tcaagtatct ggggtatctt
     7381 ctaaaagagg gtcagagatg gctgactgag gccagaaaag agactgtgat ggggcagcct
     7441 actccgaaga cccctcgaca actaagggag ttcctaggga aggcaggctt ctgtcgcctc
     7501 ttcatccctg ggtttgcaga aatggcagcc cccctgtacc ctctcaccaa accggggact
     7561 ctgtttaatt ggggcccaga ccaacaaaag gcctatcaag aaatcaagca agctcttcta
     7621 actgccccag ccctggggtt gccagatttg actaagccct ttgaactctt tgtcgacgag
     7681 aagcagggct acgccaaagg tgtcctaacg caaaaactgg gaccttggcg tcggccggtg
     7741 gcctacctgt ccaaaaagct agacccagta gcagctgggt ggcccccttg cctacggatg
     7801 gtagcagcca ttgccgtact gacaaaggat gcaggcaagc taaccatggg acagccacta
     7861 gtcattctgg ccccccatgc agtagaggca ctagtcaaac aaccccccga ccgctggctt
     7921 tccaacgccc ggatgactca ctatcaggcc ttgcttttgg acacggaccg ggtccagttc
     7981 ggaccggtgg tagccctgaa cccggctacg ctgctcccac tgcctgagga agggctgcaa
     8041 cacaactgcc ttgatatcct ggccgaagcc cacggaaccc gacccgacct aacggaccag
     8101 ccgctcccag acgccgacca cacctggtac acggatggaa gcagtctctt acaagaggga
     8161 cagcgtaagg cgggagctgc ggtgaccacc gagaccgagg taatctgggc taaagccctg
     8221 ccagccggga catccgctca gcgggctgaa ctgatagcac tcacccaggc cctaaagatg
     8281 gcagaaggta agaagctaaa tgtttatact gatagccgtt atgcttttgc tactgcccat
     8341 atccatggag aaatatacag aaggcgtggg tggctcacat cagaaggcaa agagatcaaa
     8401 aataaagacg agatcttggc cctactaaaa gccctctttc tgcccaaaag acttagcata
     8461 atccattgtc caggacatca aaagggacac agcgccgagg ctagaggcaa ccggatggct
     8521 gaccaagcgg cccgaaaggc agccatcaca gagactccag acacctctac cctcctcata
     8581 gaaaattcat caccctctgg cggctcaaaa agaaccgccg acggcagcga atttgagaag
     8641 cgacctgccg ccacaaagaa ggctggacag gctaagaaga agaaagatta caaagacgat
     8701 gacgataagg gatccggcgc aacaaacttc tctctgctga aacaagccgg agatgtcgaa
     8761 gagaatcctg gaccgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     8821 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     8881 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     8941 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     9001 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     9061 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     9121 cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga ccaccagggc
     9181 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     9241 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     9301 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     9361 cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga
     9421 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     9481 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     9541 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     9601 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     9661 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     9721 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     9781 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     9841 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     9901 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     9961 tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta
    10021 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
    10081 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
    10141 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
    10201 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
    10261 cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc gactgtgcct
    10321 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
    10381 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
    10441 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
    10501 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
    10561 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
    10621 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
    10681 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
    10741 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
    10801 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
    10861 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
    10921 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
    10981 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
    11041 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
    11101 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
    11161 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
    11221 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
    11281 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
    11341 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg
    11401 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
    11461 ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg
    11521 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
    11581 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
    11641 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
    11701 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
    11761 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
    11821 cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta
    11881 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
    11941 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
    12001 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
    12061 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
    12121 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
    12181 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
    12241 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
    12301 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
    12361 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
    12421 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
    12481 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
    12541 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
    12601 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
    12661 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
    12721 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
    12781 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
    12841 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
    12901 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
    12961 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
    13021 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
    13081 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
    13141 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
    13201 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
    13261 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
    13321 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
    13381 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
    13441 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
    13501 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
    13561 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    13621 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    13681 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    13741 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    13801 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    13861 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    13921 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    13981 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    14041 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    14101 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    14161 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    14221 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    14281 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
    14341 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
    14401 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
    14461 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
    14521 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
    14581 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
    14641 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
    14701 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
    14761 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
    14821 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
    14881 tacatctacg tattagtcat cgctattacc atgg
//
