LOCUS       sequence-176928-       13713 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modified version of lentiSAM v2, a lenti sgRNA cloning backbone with
            MS2 loops at tetraloop/stemloop 2, dCas9-VP64, and puro resistance 
            marker. Contains BsmBI sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13713)
  AUTHORS   Barger CJ, Chee L, Albahrani M, Munoz-Trujillo C, Boghean L, Branick
            C, Odunsi K, Drapkin R, Zou L, Karpf AR
  TITLE     Co-regulation and function of FOXM1/RHNO1 bidirectional genes in 
            cancer.
  JOURNAL   Elife. 2021 Apr 23;10. pii: 55070. doi: 10.7554/eLife.55070.
  PUBMED    33890574
REFERENCE   2  (bases 1 to 13713)
  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"; journalName: "Elife."; 
            date: "2021-04-23"; volume: "10. pii: 55070. doi"; pages: " 
            10.7554/eLife.55070"
COMMENT     Sequence Label: lentiSAM v2 (Puro)
FEATURES             Location/Qualifiers
     source          1..13713
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2607..2855
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2607..2627
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2778..2797
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2893..2911
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        2963..2981
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        3068..3279
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3296..3343
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3344..3367,3368..3370,3371..5926,5927..5929,
                     5930..7444)
                     /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 N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7499..7519
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7544..7693
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             7715..7771
                     /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             7772..8368
                     /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     8265..8285
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8405..8993
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8458..8478)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8876..8887)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8996..9012)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8997..9013)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9518..9698
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9724..9741)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9730..9954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10000..10428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10087..10106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10297..10318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10437..10457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10442..10771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10622..10757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10684..10703
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10819..10866
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10885..11259
                     /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    11389..11510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11426..11445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11480..11499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11559..11575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11559..11575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11572..11594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11583..11599
                     /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(11607..11613,11614..11631,11632..11637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11652..11673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11790..11807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11961..12549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12041..12060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12720..13511,13512..13580))
                     /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     13343..13362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13581..13685)
                     /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 ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta gctagcgagg gcctatttcc catgattcct tcatatttgc
     2641 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
     2701 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     2761 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     2821 tggctttata tatcttgtgg aaaggacgaa acaccggaga cgggataccg tctctgtttt
     2881 agagctaggc caacatgagg atcacccatg tctgcagggc ctagcaagtt aaaataaggc
     2941 tagtccgtta tcaacttggc caacatgagg atcacccatg tctgcagggc caagtggcac
     3001 cgagtcggtg ctttttttgg atcctaggtc ttgaaaggag tgggaattgg ctccggtgcc
     3061 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     3121 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     3181 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     3241 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg cgtacggcca ccatgaaaag
     3301 gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aaggacaaga agtacagcat
     3361 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
     3421 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
     3481 gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac
     3541 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
     3601 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
     3661 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
     3721 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
     3781 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
     3841 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
     3901 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
     3961 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
     4021 ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg
     4081 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
     4141 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
     4201 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
     4261 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
     4321 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
     4381 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc
     4441 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
     4501 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
     4561 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
     4621 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
     4681 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
     4741 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc
     4801 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
     4861 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt
     4921 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc
     4981 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
     5041 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
     5101 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
     5161 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
     5221 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
     5281 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
     5341 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
     5401 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
     5461 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
     5521 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
     5581 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
     5641 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
     5701 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
     5761 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
     5821 actggacatc aaccggctgt ccgactacga tgtggaccac atcgtgcctc agagctttct
     5881 gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaaggccc ggggcaagag
     5941 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct
     6001 gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg
     6061 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
     6121 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga
     6181 caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
     6241 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
     6301 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
     6361 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
     6421 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
     6481 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
     6541 cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct
     6601 gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
     6661 agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga
     6721 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
     6781 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
     6841 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
     6901 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
     6961 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
     7021 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
     7081 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga
     7141 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
     7201 ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga
     7261 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
     7321 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
     7381 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
     7441 cgacagcgct ggaggaggtg gaagcggagg aggaggaagc ggaggaggag gtagcggacc
     7501 taagaaaaag aggaaggtgg cggccgctgg atccggacgg gctgacgcat tggacgattt
     7561 tgatctggat atgctgggaa gtgacgccct cgatgatttt gaccttgaca tgcttggttc
     7621 ggatgccctt gatgactttg acctcgacat gctcggcagt gacgcccttg atgatttcga
     7681 cctggacatg ctgattaact gtacaggcag tggagcaaca aacttctctc tgctgaaaca
     7741 agccggagat gtcgaagaga atcctggacc gaccgagtac aagcccacgg tgcgcctcgc
     7801 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     7861 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     7921 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     7981 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     8041 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     8101 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc
     8161 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     8221 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     8281 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     8341 gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc gaattcgata tcaagcttat
     8401 cggtaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     8461 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     8521 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     8581 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     8641 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     8701 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     8761 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     8821 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     8881 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     8941 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     9001 ccgtcgacct cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
     9061 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
     9121 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     9181 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     9241 tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc
     9301 agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga
     9361 gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca
     9421 tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt
     9481 tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat
     9541 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     9601 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     9661 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
     9721 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     9781 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     9841 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     9901 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     9961 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
    10021 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
    10081 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
    10141 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
    10201 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
    10261 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
    10321 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
    10381 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
    10441 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
    10501 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
    10561 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
    10621 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
    10681 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
    10741 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
    10801 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
    10861 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
    10921 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
    10981 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
    11041 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
    11101 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
    11161 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
    11221 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
    11281 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
    11341 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
    11401 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
    11461 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
    11521 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
    11581 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
    11641 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
    11701 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
    11761 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
    11821 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
    11881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
    11941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
    12001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
    12061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
    12121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
    12181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
    12241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
    12301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
    12361 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
    12421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
    12481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
    12541 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
    12601 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
    12661 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
    12721 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
    12781 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    12841 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
    12901 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    12961 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    13021 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    13081 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    13141 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    13201 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    13261 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    13321 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    13381 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    13441 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    13501 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    13561 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    13621 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    13681 ccgcgcacat ttccccgaaa agtgccacct gac
//
