LOCUS       sequence-325014-       13264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Makes a lentivirus that expresses mNeonGreen and Cas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13264)
  AUTHORS   Loveless TB, Grotts JH, Schechter MW, Forouzmand E, Carlson CK, 
            Agahi BS, Liang G, Ficht M, Liu B, Xie X, Liu CC
  TITLE     Lineage tracing and analog recording in mammalian cells by 
            single-site DNA writing.
  JOURNAL   Nat Chem Biol. 2021 Jun;17(6):739-747. doi: 
            10.1038/s41589-021-00769-8. Epub 2021 Mar 22.
  PUBMED    33753928
REFERENCE   2  (bases 1 to 13264)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41589-021-00769-8"; journalName: "Nat Chem Biol"; date: 
            "2021-06"; volume: "17"; issue: "6"; pages: "739-747"
COMMENT     Sequence Label: pTBL1401 pLenti-pEF1a(short)-mNeonGreen-P2A-Cas9
FEATURES             Location/Qualifiers
     source          1..13264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2352..2469
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2538..2749
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2769..2778
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3543..3599
                     /label=MCS
                     /note="multiple cloning site"
     CDS             3621..3677
                     /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             3681..3704
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3711..3731
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3756..7856
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3756..7856
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7857..7904
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7905..7928
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    7956..8544
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8009..8029)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8427..8438)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8547..8563)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8548..8564)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9069..9249
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9275..9292)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9281..9505
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9551..9979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9638..9657)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9848..9869
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9988..10008)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9993..10322
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10173..10308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10235..10254
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10370..10417
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10436..10810
                     /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    10940..11061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10977..10996)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11031..11050
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11110..11126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11110..11126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11123..11145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11134..11150
                     /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(11158..11164,11165..11182,11183..11188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11203..11224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11341..11358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11512..12100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11592..11611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12271..13062,13063..13131))
                     /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     12894..12913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13132..13236)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggaccctaga tcttgagaca aatggcagta
     2341 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     2401 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     2461 caaaattttc gggtttatta cagggacagc agagatccac tttggcgccg gctcgagtgg
     2521 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     2581 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     2641 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     2701 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg tgtcgtgacg
     2761 cgggatccgc caccatggtc agcaaaggcg aagaagacaa catggcatca ctgccagcga
     2821 cacacgaact tcatattttc ggctccatca atggtgtgga cttcgatatg gttggacagg
     2881 gtactgggaa ccctaacgat ggatatgagg agttgaactt gaagagtacc aagggcgatc
     2941 ttcagttttc accttggatt ctcgtacctc atataggtta tggctttcat caatatctgc
     3001 cctatcctga cggtatgtca cctttccaag cagctatggt tgacgggagc ggttatcagg
     3061 ttcaccggac aatgcagttt gaagatggag cctctttgac cgtaaattat cggtatactt
     3121 atgagggcag ccatattaaa ggcgaggctc aagttaaggg gactgggttc cccgcggatg
     3181 gtcccgtaat gacaaacagt ctgacggctg cagactggtg ccgctctaag aagacatatc
     3241 caaatgacaa gacgattatt agcactttca agtggtcata cactacagga aatggtaaac
     3301 gctataggag cactgcccgg accacgtata ctttcgcgaa gccgatggcg gcaaattatt
     3361 tgaaaaacca accgatgtat gtgttcagaa agacagaact caagcattct aaaactgaac
     3421 tgaacttcaa agagtggcag aaagccttta ctgatgtaat gggcatggac gaactctata
     3481 agtccggact cagatccggt ggaggcggtt caggcggagg tggctctggc ggtggcggat
     3541 cgagatctcg agctcaagct tcgaactctg cagtcgacgg taccgcgggc ccgggatcca
     3601 ccggatctag aggatccggc gcaacaaact tctctctgct gaaacaagcc ggagatgtcg
     3661 aagagaatcc tggaccgatg gattacaaag acgatgacga taagatggcc ccaaagaaga
     3721 agcggaaggt cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg
     3781 acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca
     3841 agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag
     3901 ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa
     3961 gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga
     4021 tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg
     4081 ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg
     4141 agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg
     4201 acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga
     4261 tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc
     4321 agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg
     4381 ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc
     4441 ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc
     4501 ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca
     4561 cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt
     4621 ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca
     4681 ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc
     4741 aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga
     4801 ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg
     4861 aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc
     4921 tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca
     4981 tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt
     5041 acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct
     5101 actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg
     5161 aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga
     5221 gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca
     5281 agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg
     5341 tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg
     5401 acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca
     5461 agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct
     5521 ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg
     5581 aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag
     5641 agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc
     5701 agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca
     5761 tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca
     5821 acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga
     5881 aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca
     5941 gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag
     6001 tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca
     6061 cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag
     6121 agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga
     6181 agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca
     6241 tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg
     6301 actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg
     6361 tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca
     6421 agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg
     6481 aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc
     6541 acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga
     6601 tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt
     6661 tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga
     6721 acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt
     6781 acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg
     6841 gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga
     6901 ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg
     6961 gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc
     7021 cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta
     7081 tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga
     7141 agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg
     7201 aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg
     7261 aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag
     7321 tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc
     7381 ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct
     7441 ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg
     7501 aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca
     7561 tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag
     7621 tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca
     7681 tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca
     7741 ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc
     7801 agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaagc
     7861 gacctgccgc cacaaagaag gctggacagg ctaagaagaa gaaagattac aaagacgatg
     7921 acgataagta acaattcgat atcaagctta tcggtaatca acctctggat tacaaaattt
     7981 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     8041 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     8101 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     8161 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     8221 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     8281 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     8341 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     8401 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     8461 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     8521 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta gaaaaacatg
     8581 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
     8641 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
     8701 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     8761 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
     8821 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat
     8881 ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga
     8941 acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat
     9001 tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg
     9061 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     9121 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     9181 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     9241 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     9301 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     9361 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     9421 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     9481 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     9541 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     9601 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     9661 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     9721 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     9781 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     9841 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     9901 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     9961 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
    10021 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
    10081 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
    10141 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
    10201 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
    10261 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
    10321 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
    10381 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
    10441 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
    10501 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
    10561 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
    10621 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
    10681 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
    10741 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
    10801 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
    10861 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
    10921 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
    10981 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    11041 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
    11101 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
    11161 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
    11221 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
    11281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
    11341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
    11401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
    11461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
    11521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
    11581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
    11641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
    11701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    11761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    11821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    11881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    11941 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    12001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    12061 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    12121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
    12181 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
    12241 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
    12301 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
    12361 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
    12421 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
    12481 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
    12541 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    12601 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    12661 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    12721 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    12781 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    12841 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    12901 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    12961 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    13021 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    13081 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    13141 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    13201 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    13261 tgac
//
