LOCUS       sequence-256506-       12103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6 promoter expresses customizable Spyo-guide; EFS promoter 
            expresses SaurCas9 and 2A site provides puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12103)
  AUTHORS   DeWeirdt PC, Sangree AK, Hanna RE, Sanson KR, Hegde M, Strand C, 
            Persky NS, Doench JG
  TITLE     Genetic screens in isogenic mammalian cell lines without single cell
            cloning.
  JOURNAL   Nat Commun. 2020 Feb 6;11(1):752. doi: 10.1038/s41467-020-14620-6.
  PUBMED    32029722
REFERENCE   2  (bases 1 to 12103)
  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: "Nat 
            Commun."; date: "2020-02-6"; volume: "11(1):752. doi"; pages: " 
            10.1038/s41467-020-14620-6"
COMMENT     Sequence Label: pXPR_212
FEATURES             Location/Qualifiers
     source          1..12103
                     /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        2608..2856
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2608..2628
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2779..2798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2878..2953
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3015..3226
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3234..3243
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3246..3266
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3291..6446
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
     CDS             6447..6494
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6504..6560
                     /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"
     misc_feature    7189..7777
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7242..7262)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7660..7671)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7855..8088
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8114..8131)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8120..8344
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8390..8818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8477..8496)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8687..8708
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8827..8847)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8832..9161
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9012..9147
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9074..9093
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9209..9256
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9275..9649
                     /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    9779..9900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9816..9835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9870..9889
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9949..9965)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9949..9965)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9962..9984)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9973..9989
                     /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(9997..10003,10004..10021,10022..10027))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10042..10063
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10180..10197)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10351..10939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10431..10450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11110..11901,11902..11970))
                     /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     11733..11752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11971..12075)
                     /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 aggtaccgag ggcctatttc ccatgattcc ttcatatttg
     2641 catatacgat acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccggag acgctttttt cgtctcagtt
     2881 tgagagctag aaatagcaag ttcaaataag gctagtccgt tatcaacttg aaaaagtggc
     2941 accgagtcgg tgcttttttg aattcgctag ctaggtcttg aaaggagtgg gaattggctc
     3001 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     3061 ggtcggcaat tgatccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     3121 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     3181 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggacc ggtgccacca
     3241 tggcacccaa gaagaagcgc aaagttggca tccacggcgt gcctgctgcc aagcgcaact
     3301 acatcctggg cctggacatt ggcatcacct cagttggcta tggcatcatc gactacgaga
     3361 cccgggacgt catcgacgct ggcgtccgcc tcttcaagga ggccaacgtc gagaacaacg
     3421 agggccggcg gtccaagcgc ggggcccgcc ggctcaagcg ccgccgccgc caccgcatcc
     3481 agcgcgtgaa aaagttgctc ttcgactaca acttgctcac cgaccactcc gagctgtctg
     3541 gcatcaaccc gtacgaggcc cgggtcaagg gcctctcgca aaagttgtcc gaggaggagt
     3601 tctctgctgc cctgctgcac ctggccaagc gccgcggggt ccacaacgtc aacgaagttg
     3661 aggaggacac cggcaacgaa ctttccacca aggagcagat ctcccgcaac tccaaggccc
     3721 tggaggagaa gtacgtcgcg gagctccaac ttgagcgcct caagaaggac ggcgaggtgc
     3781 gcggctccat caaccgcttc aagacctccg actacgtcaa ggaggccaag cagctgctca
     3841 aggtccagaa ggcctaccac caacttgacc agtccttcat cgacacctac atcgacctgc
     3901 tggagacccg ccgcacctac tacgaggggc ctggcgaggg ctctcccttc ggctggaagg
     3961 acatcaagga gtggtacgag atgctcatgg gccactgcac ctacttcccg gaggagctgc
     4021 gctcggtcaa gtacgcctac aacgccgacc tctacaacgc gctcaacgac ctcaacaact
     4081 tggtcatcac ccgcgacgag aacgaaaagt tggagtacta cgaaaagttc cagatcatcg
     4141 aaaacgtctt caagcagaag aagaagccaa ctttgaagca gatcgccaag gagatcctgg
     4201 tcaacgagga ggacatcaag ggctaccggg tcacctccac cggcaagccc gagttcacca
     4261 acttaaaggt ctaccatgac atcaaggaca tcaccgcccg caaggagatc atcgagaacg
     4321 cggagctgct ggaccagatc gccaagatcc tcaccatcta ccaaagttcc gaggacatcc
     4381 aggaggagct gaccaactta aactcggagc ttacccagga ggagatcgag cagatctcca
     4441 acttaaaggg ctacactggc acccacaact tgtccctcaa ggccatcaac ttgatcctgg
     4501 acgaactttg gcataccaac gacaaccaga tcgccatctt caaccgacta aagttggtgc
     4561 ccaagaaagt tgacctgtcc caacagaagg agatcccaac taccctggtc gacgacttca
     4621 tcctctcgcc cgtggtcaag cgcagcttca tccagtccat caaggtcatc aacgccatca
     4681 tcaagaagta cggcctgccc aacgacatca tcatcgagtt ggcacgcgag aagaactcca
     4741 aggatgccca gaagatgatc aacgagatgc agaagcgcaa ccgccagacc aacgagcgca
     4801 tcgaggagat catccgcacc accggcaagg agaacgccaa gtacctgatc gagaagataa
     4861 agttgcatga catgcaggag ggcaagtgcc tctactccct ggaggccatc cccctcgagg
     4921 acctgctcaa caaccccttc aactacgaag ttgaccacat catcccgcgc tcggtgtcct
     4981 tcgacaacag cttcaacaac aaggtcctgg tcaagcagga ggagaactcc aagaagggca
     5041 accggacccc cttccagtac ctgtcctcct cggactccaa gatctcctac gaaactttca
     5101 agaagcacat cctcaacttg gcaaagggca agggccgcat ctccaagacc aagaaggagt
     5161 acctgctgga ggagcgcgac atcaaccgct tctcggtgca gaaggacttc atcaaccgca
     5221 acctggtgga cacccgctac gcaacccgtg gcctcatgaa cctgctgcgc agctacttcc
     5281 gggtcaacaa cttggacgtc aaagttaagt cgatcaacgg tggcttcacc agcttcctgc
     5341 gccgcaagtg gaagttcaag aaggagcgca acaagggcta caagcaccat gcggaggacg
     5401 ccctcatcat cgccaacgcc gacttcatct tcaaggagtg gaaaaagttg gacaaggcca
     5461 agaaggtcat ggagaaccag atgttcgagg agaagcaggc cgagtccatg ccggagatcg
     5521 agacagagca ggagtacaag gagatcttca tcacgccgca ccagatcaag cacatcaagg
     5581 acttcaagga ctacaagtac tcccaccgcg tggacaagaa gcccaaccgc gagctcatca
     5641 acgacaccct ctactccacc cgcaaggacg acaagggcaa caccctcatc gtcaacaact
     5701 taaacggcct ctacgacaag gacaacgaca agttgaaaaa gttgatcaac aagagccccg
     5761 aaaagttgct catgtaccac catgacccgc agacgtacca aaagttgaag ttgatcatgg
     5821 agcagtacgg tgacgagaag aacccgctct acaagtacta cgaggagacc ggcaactacc
     5881 tcaccaagta ctccaagaag gacaacggcc cggtcatcaa gaagatcaag tactacggca
     5941 acaagttgaa tgcccacctg gacatcaccg acgactaccc caactcccgc aacaaagttg
     6001 taaagttgtc gctaaagccg taccgcttcg acgtctacct ggacaacggc gtctacaagt
     6061 tcgtgaccgt caagaacctg gacgtcatca agaaggagaa ctactacgag gtcaactcca
     6121 agtgctacga ggaggccaaa aagttgaaga agatctccaa ccaggccgag ttcatcgcca
     6181 gcttctacaa caacgacctc atcaagatca acggcgagct ctaccgcgtc atcggggtca
     6241 acaacgacct gctgaaccgc atcgaggtca acatgatcga catcacctac cgcgagtacc
     6301 tcgagaacat gaacgacaag cgcccgcctc gcatcatcaa gaccatcgcc tccaagaccc
     6361 agtccatcaa gaagtactcc accgacatcc ttggcaactt gtacgaggtc aagagcaaga
     6421 agcaccctca gatcatcaag aagggcaagc gccctgctgc caccaagaag gctggccagg
     6481 ccaagaagaa gaagggatcc ggggccacca acttttccct gctgaagcag gctggcgacg
     6541 tcgaggagaa ccctggccct ggctccggaa ccgagtacaa gccaactgtc cgcttggcaa
     6601 cccgcgacga tgtgcccagg gccgtccgca cgctggctgc tgccttcgcc gactacccag
     6661 ccacccgcca caccgtggac ccggaccgcc acatcgagcg cgtcaccgag ctgcaggagc
     6721 tcttcctcac ccgagttggc cttgacattg gcaaggtctg ggtggcggac gacggtgctg
     6781 ccgtggccgt ctggaccacc cccgagtcgg tcgaggctgg cgccgtcttc gccgagatcg
     6841 ggccgcgcat ggccgagctt tcgggctccc ggttggctgc ccaacagcag atggaggggc
     6901 tgttggcacc ccaccgcccc aaggagccgg cctggttcct ggccacagtt ggtgtctctc
     6961 ccgaccacca gggcaagggc ctgggcagcg ccgtggtgct gcctggggtc gaggctgccg
     7021 agcgcgctgg cgtccctgcc ttcctggaga cctctgcccc tcgcaacttg cccttctacg
     7081 agcgcctggg cttcaccgtc accgcggacg tcgaggtgcc cgaggggccg cgcacctggt
     7141 gcatgacccg caagcctggg gcatgaatct agaacgcgtt aagtcgacaa tcaacctctg
     7201 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     7261 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     7321 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     7381 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     7441 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     7501 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     7561 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     7621 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     7681 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     7741 cagacgagtc ggatctccct ttgggccgcc tccccgcagc gctgtcgact ttaagaccaa
     7801 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     7861 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
     7921 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     7981 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     8041 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa
     8101 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     8161 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     8221 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     8281 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     8341 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     8401 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     8461 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     8521 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     8581 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     8641 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     8701 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     8761 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     8821 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     8881 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     8941 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     9001 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     9061 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     9121 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     9181 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     9241 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
     9301 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     9361 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     9421 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     9481 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     9541 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     9601 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg
     9661 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     9721 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     9781 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     9841 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     9901 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     9961 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
    10021 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
    10081 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
    10141 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
    10201 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
    10261 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
    10321 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
    10381 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
    10441 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
    10501 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
    10561 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
    10621 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
    10681 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
    10741 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
    10801 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
    10861 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
    10921 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
    10981 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
    11041 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
    11101 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
    11161 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
    11221 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
    11281 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
    11341 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
    11401 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
    11461 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
    11521 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
    11581 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
    11641 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
    11701 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
    11761 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
    11821 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
    11881 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    11941 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    12001 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    12061 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gac
//
