LOCUS       Exported               14987 bp ds-DNA     circular SYN 25-MAR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14987)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Dec 9, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14987
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2364..2397
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_RNA        complement(2408..2479)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(2513..2753)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(2563..2582)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2733..2753)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        2786..3997
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3834..3853
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      4026..4035
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4035..4100
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4107..4127
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4137..8240
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8253..8273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8307..8501
                     /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             8568..8588
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8595..8648
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             8655..9368
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             8655..9368
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(8697..8718)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8958..8977)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9305..9326
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    9384..9417
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    9442..10030
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9495..9515)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9913..9924)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10033..10049)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10034..10050)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10555..10735
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10761..10778)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10767..10991
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11037..11465
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11124..11143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11334..11355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11474..11494)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11479..11808
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11659..11794
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11721..11740
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11856..11903
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11922..12296
                     /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    12426..12547
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12463..12482)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12517..12536
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12596..12612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12596..12612)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12609..12631)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12620..12636
                     /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(12644..12674)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12689..12710
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12827..12844)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12998..13586)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13078..13097)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13757..14617)
                     /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     14380..14399
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14618..14722)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14797..14816)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaataactt cgtatagcat acattatacg aagttattaa
     2401 aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc cttattttaa
     2461 cttgctattt ctagctctaa aactcaccac gtaggagttc ggccggtgtt tcgtcctttc
     2521 cacaagatat ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag
     2581 tccattttaa aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt
     2641 cacgtatttt gtactaatat ctttgtgttt acagtcaaat taattccaat tatctctcta
     2701 acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcttaatta
     2761 attaacccgt gtcggctcca gatctggcct ccgcgccggg ttttggcgcc tcccgcgggc
     2821 gcccccctcc tcacggcgag cgctgccacg tcagacgaag ggcgcagcga gcgtcctgat
     2881 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
     2941 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
     3001 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
     3061 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
     3121 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
     3181 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
     3241 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     3301 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     3361 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     3421 caagaaccca aggtcttgag gccttcgcta atgcgggaaa gctcttattc gggtgagatg
     3481 ggctggggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     3541 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     3601 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     3661 gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     3721 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     3781 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     3841 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     3901 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagacta gtaaattgtc
     3961 cgctaaattc tggccgtttt tggctttttt gttagacgaa gcttgggctg caggtcgact
     4021 ctagagccac catggactac aaagaccatg acggtgatta taaagatcat gacatcgatt
     4081 acaaggatga cgatgacaag atggccccca agaagaagag gaaggtgggc cgcggaatgg
     4141 acaagaagta ctccattggg ctcgccatcg gcacaaacag cgtcggctgg gccgtcatta
     4201 cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc gatcgccaca
     4261 gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggaaacc gccgaagcca
     4321 cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg atctgctacc
     4381 tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc cataggctgg
     4441 aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc tttggcaata
     4501 tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg aggaagaagc
     4561 ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg gcgcatatga
     4621 tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac agcgatgtcg
     4681 acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag aacccgatca
     4741 acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc
     4801 tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt ggtaatctta
     4861 tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg gccgaagatg
     4921 ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg ctggcccaga
     4981 tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac gccattctgc
     5041 tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc gctagtatga
     5101 tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt gtcagacagc
     5161 aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc tacgccggat
     5221 acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc atcttggaaa
     5281 aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg ttgcgcaaac
     5341 agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa ctgcacgcta
     5401 tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa aagattgaga
     5461 aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga aattccagat
     5521 tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc gaggaagtcg
     5581 tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt gataaaaatc
     5641 tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc acagtttata
     5701 acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca ttcctgtctg
     5761 gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa gttaccgtga
     5821 aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg
     5881 gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg aaaatcatta
     5941 aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac attgtcctca
     6001 cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact tacgctcatc
     6061 tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga tgggggcggc
     6121 tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca atcctggatt
     6181 ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat gatgactctc
     6241 tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac agtcttcacg
     6301 agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg cagaccgtta
     6361 aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat atcgttatcg
     6421 agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg gaaaggatga
     6481 agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa cacccagttg
     6541 aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac ggcagggaca
     6601 tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg gatgccatcg
     6661 tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca agatccgata
     6721 aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt
     6781 attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat aatctgacta
     6841 aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa aggcagcttg
     6901 ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc atgaacacca
     6961 agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg aagtctaagc
     7021 tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc aacaattacc
     7081 accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc
     7141 ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt aggaaaatga
     7201 tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt tacagcaata
     7261 ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg aagcgaccac
     7321 ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg gatttcgcga
     7381 cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc gaagtacaga
     7441 ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac
     7501 gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca gtcgcttaca
     7561 gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg
     7621 aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc atcgactttc
     7681 tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt cccaagtact
     7741 ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga
     7801 aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg gccagccact
     7861 atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc gtggaacaac
     7921 acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa agagtgatcc
     7981 tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg gataagccca
     8041 tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg ggcgcgcctg
     8101 cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct acaaaggagg
     8161 tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca agaatcgacc
     8221 tctctcagct cggtggagac agcagggctg accccaagaa gaagaggaag gtggctagcg
     8281 atgctaagtc actgactgcc tggtcccgga cactggtgac cttcaaggat gtgtttgtgg
     8341 acttcaccag ggaggagtgg aagctgctgg acactgctca gcagatcctg tacagaaatg
     8401 tgatgctgga gaactataag aacctggttt ccttgggtta tcagcttact aagccagatg
     8461 tgatcctccg gttggagaag ggagaagagc cctggctggt ggagagagaa attcaccaag
     8521 agacccatcc tgattcagag actgcatttg aaatcaaatc atcagttccg aaaaagaaac
     8581 gcaaagttgc tagcgagggc agaggaagtc ttctaacatg cggtgacgtg gaggagaatc
     8641 ccggccctgg taccgtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     8701 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     8761 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     8821 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     8881 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     8941 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     9001 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     9061 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     9121 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     9181 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     9241 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     9301 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     9361 tgtacaagac cggttgataa tagataactt cgtatagcat acattatacg aagttatgaa
     9421 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     9481 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     9541 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     9601 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     9661 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     9721 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     9781 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     9841 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     9901 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     9961 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
    10021 gcctccccgc atcgataccg tcgacctcga gacctagaaa aacatggagc aatcacaagt
    10081 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
    10141 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
    10201 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
    10261 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag
    10321 aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta
    10381 gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta
    10441 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt
    10501 gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg tactgggtct
    10561 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
    10621 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
    10681 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagggcc
    10741 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
    10801 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
    10861 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
    10921 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
    10981 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
    11041 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
    11101 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
    11161 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
    11221 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
    11281 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
    11341 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
    11401 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
    11461 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
    11521 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
    11581 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
    11641 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
    11701 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
    11761 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
    11821 gcttgtatat ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt
    11881 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
    11941 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
    12001 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
    12061 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
    12121 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
    12181 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
    12241 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
    12301 tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
    12361 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
    12421 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
    12481 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
    12541 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
    12601 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
    12661 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
    12721 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
    12781 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
    12841 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
    12901 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
    12961 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
    13021 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
    13081 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
    13141 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
    13201 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
    13261 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
    13321 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
    13381 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
    13441 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
    13501 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
    13561 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
    13621 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
    13681 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
    13741 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
    13801 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
    13861 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
    13921 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
    13981 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
    14041 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
    14101 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
    14161 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
    14221 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
    14281 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
    14341 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
    14401 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
    14461 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
    14521 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
    14581 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
    14641 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
    14701 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
    14761 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
    14821 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
    14881 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
    14941 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
