LOCUS       sequence-261407-       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"
     promoter        2..200
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             218..398
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    445..570
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1063..1296
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1481..1525
                     /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    1823..1940
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1991..2239
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1991..2011
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2162..2181
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2261..2336
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2398..2609
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2617..2626
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2629..2649
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2674..5829
                     /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             5830..5877
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5887..5943
                     /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    6572..7160
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6625..6645)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7043..7054)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7238..7471
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7497..7514)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7503..7727
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7773..8201
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7860..7879)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8070..8091
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8210..8230)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8215..8544
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8395..8530
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8457..8476
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8592..8639
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8658..9032
                     /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    9162..9283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9199..9218)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9253..9272
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9332..9348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9332..9348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9345..9367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9356..9372
                     /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(9380..9386,9387..9404,9405..9410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9425..9446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9563..9580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9734..10322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9814..9833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10493..11284,11285..11353))
                     /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     11116..11135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11354..11458)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11533..11552)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11724..12103
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
      241 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
      301 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
      361 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
      421 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
      481 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
      541 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
      601 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
      661 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
      721 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
      781 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
      841 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
      901 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
      961 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1021 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1081 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1141 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1201 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1261 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1321 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1381 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1441 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1501 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1561 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1621 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1681 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1741 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     1801 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     1861 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     1921 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa
     1981 ttaaggtacc gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     2041 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     2101 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     2161 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     2221 gtggaaagga cgaaacaccg gagacgcttt tttcgtctca gtttgagagc tagaaatagc
     2281 aagttcaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     2341 ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     2401 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     2461 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     2521 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     2581 ttcgcaacgg gtttgccgcc agaacacagg accggtgcca ccatggcacc caagaagaag
     2641 cgcaaagttg gcatccacgg cgtgcctgct gccaagcgca actacatcct gggcctggac
     2701 attggcatca cctcagttgg ctatggcatc atcgactacg agacccggga cgtcatcgac
     2761 gctggcgtcc gcctcttcaa ggaggccaac gtcgagaaca acgagggccg gcggtccaag
     2821 cgcggggccc gccggctcaa gcgccgccgc cgccaccgca tccagcgcgt gaaaaagttg
     2881 ctcttcgact acaacttgct caccgaccac tccgagctgt ctggcatcaa cccgtacgag
     2941 gcccgggtca agggcctctc gcaaaagttg tccgaggagg agttctctgc tgccctgctg
     3001 cacctggcca agcgccgcgg ggtccacaac gtcaacgaag ttgaggagga caccggcaac
     3061 gaactttcca ccaaggagca gatctcccgc aactccaagg ccctggagga gaagtacgtc
     3121 gcggagctcc aacttgagcg cctcaagaag gacggcgagg tgcgcggctc catcaaccgc
     3181 ttcaagacct ccgactacgt caaggaggcc aagcagctgc tcaaggtcca gaaggcctac
     3241 caccaacttg accagtcctt catcgacacc tacatcgacc tgctggagac ccgccgcacc
     3301 tactacgagg ggcctggcga gggctctccc ttcggctgga aggacatcaa ggagtggtac
     3361 gagatgctca tgggccactg cacctacttc ccggaggagc tgcgctcggt caagtacgcc
     3421 tacaacgccg acctctacaa cgcgctcaac gacctcaaca acttggtcat cacccgcgac
     3481 gagaacgaaa agttggagta ctacgaaaag ttccagatca tcgaaaacgt cttcaagcag
     3541 aagaagaagc caactttgaa gcagatcgcc aaggagatcc tggtcaacga ggaggacatc
     3601 aagggctacc gggtcacctc caccggcaag cccgagttca ccaacttaaa ggtctaccat
     3661 gacatcaagg acatcaccgc ccgcaaggag atcatcgaga acgcggagct gctggaccag
     3721 atcgccaaga tcctcaccat ctaccaaagt tccgaggaca tccaggagga gctgaccaac
     3781 ttaaactcgg agcttaccca ggaggagatc gagcagatct ccaacttaaa gggctacact
     3841 ggcacccaca acttgtccct caaggccatc aacttgatcc tggacgaact ttggcatacc
     3901 aacgacaacc agatcgccat cttcaaccga ctaaagttgg tgcccaagaa agttgacctg
     3961 tcccaacaga aggagatccc aactaccctg gtcgacgact tcatcctctc gcccgtggtc
     4021 aagcgcagct tcatccagtc catcaaggtc atcaacgcca tcatcaagaa gtacggcctg
     4081 cccaacgaca tcatcatcga gttggcacgc gagaagaact ccaaggatgc ccagaagatg
     4141 atcaacgaga tgcagaagcg caaccgccag accaacgagc gcatcgagga gatcatccgc
     4201 accaccggca aggagaacgc caagtacctg atcgagaaga taaagttgca tgacatgcag
     4261 gagggcaagt gcctctactc cctggaggcc atccccctcg aggacctgct caacaacccc
     4321 ttcaactacg aagttgacca catcatcccg cgctcggtgt ccttcgacaa cagcttcaac
     4381 aacaaggtcc tggtcaagca ggaggagaac tccaagaagg gcaaccggac ccccttccag
     4441 tacctgtcct cctcggactc caagatctcc tacgaaactt tcaagaagca catcctcaac
     4501 ttggcaaagg gcaagggccg catctccaag accaagaagg agtacctgct ggaggagcgc
     4561 gacatcaacc gcttctcggt gcagaaggac ttcatcaacc gcaacctggt ggacacccgc
     4621 tacgcaaccc gtggcctcat gaacctgctg cgcagctact tccgggtcaa caacttggac
     4681 gtcaaagtta agtcgatcaa cggtggcttc accagcttcc tgcgccgcaa gtggaagttc
     4741 aagaaggagc gcaacaaggg ctacaagcac catgcggagg acgccctcat catcgccaac
     4801 gccgacttca tcttcaagga gtggaaaaag ttggacaagg ccaagaaggt catggagaac
     4861 cagatgttcg aggagaagca ggccgagtcc atgccggaga tcgagacaga gcaggagtac
     4921 aaggagatct tcatcacgcc gcaccagatc aagcacatca aggacttcaa ggactacaag
     4981 tactcccacc gcgtggacaa gaagcccaac cgcgagctca tcaacgacac cctctactcc
     5041 acccgcaagg acgacaaggg caacaccctc atcgtcaaca acttaaacgg cctctacgac
     5101 aaggacaacg acaagttgaa aaagttgatc aacaagagcc ccgaaaagtt gctcatgtac
     5161 caccatgacc cgcagacgta ccaaaagttg aagttgatca tggagcagta cggtgacgag
     5221 aagaacccgc tctacaagta ctacgaggag accggcaact acctcaccaa gtactccaag
     5281 aaggacaacg gcccggtcat caagaagatc aagtactacg gcaacaagtt gaatgcccac
     5341 ctggacatca ccgacgacta ccccaactcc cgcaacaaag ttgtaaagtt gtcgctaaag
     5401 ccgtaccgct tcgacgtcta cctggacaac ggcgtctaca agttcgtgac cgtcaagaac
     5461 ctggacgtca tcaagaagga gaactactac gaggtcaact ccaagtgcta cgaggaggcc
     5521 aaaaagttga agaagatctc caaccaggcc gagttcatcg ccagcttcta caacaacgac
     5581 ctcatcaaga tcaacggcga gctctaccgc gtcatcgggg tcaacaacga cctgctgaac
     5641 cgcatcgagg tcaacatgat cgacatcacc taccgcgagt acctcgagaa catgaacgac
     5701 aagcgcccgc ctcgcatcat caagaccatc gcctccaaga cccagtccat caagaagtac
     5761 tccaccgaca tccttggcaa cttgtacgag gtcaagagca agaagcaccc tcagatcatc
     5821 aagaagggca agcgccctgc tgccaccaag aaggctggcc aggccaagaa gaagaaggga
     5881 tccggggcca ccaacttttc cctgctgaag caggctggcg acgtcgagga gaaccctggc
     5941 cctggctccg gaaccgagta caagccaact gtccgcttgg caacccgcga cgatgtgccc
     6001 agggccgtcc gcacgctggc tgctgccttc gccgactacc cagccacccg ccacaccgtg
     6061 gacccggacc gccacatcga gcgcgtcacc gagctgcagg agctcttcct cacccgagtt
     6121 ggccttgaca ttggcaaggt ctgggtggcg gacgacggtg ctgccgtggc cgtctggacc
     6181 acccccgagt cggtcgaggc tggcgccgtc ttcgccgaga tcgggccgcg catggccgag
     6241 ctttcgggct cccggttggc tgcccaacag cagatggagg ggctgttggc accccaccgc
     6301 cccaaggagc cggcctggtt cctggccaca gttggtgtct ctcccgacca ccagggcaag
     6361 ggcctgggca gcgccgtggt gctgcctggg gtcgaggctg ccgagcgcgc tggcgtccct
     6421 gccttcctgg agacctctgc ccctcgcaac ttgcccttct acgagcgcct gggcttcacc
     6481 gtcaccgcgg acgtcgaggt gcccgagggg ccgcgcacct ggtgcatgac ccgcaagcct
     6541 ggggcatgaa tctagaacgc gttaagtcga caatcaacct ctggattaca aaatttgtga
     6601 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     6661 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     6721 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     6781 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     6841 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     6901 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     6961 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     7021 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     7081 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     7141 cctttgggcc gcctccccgc agcgctgtcg actttaagac caatgactta caaggcagct
     7201 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     7261 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     7321 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     7381 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     7441 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
     7501 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     7561 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     7621 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     7681 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     7741 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     7801 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     7861 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     7921 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     7981 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     8041 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     8101 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     8161 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     8221 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     8281 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     8341 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     8401 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     8461 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     8521 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     8581 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     8641 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     8701 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     8761 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     8821 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     8881 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     8941 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     9001 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
     9061 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     9121 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     9181 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     9241 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     9301 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     9361 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     9421 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     9481 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     9541 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     9601 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     9661 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     9721 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     9781 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     9841 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     9901 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     9961 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    10021 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    10081 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    10141 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    10201 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    10261 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    10321 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    10381 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    10441 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    10501 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    10561 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    10621 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    10681 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    10741 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    10801 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    10861 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    10921 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    10981 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    11041 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    11101 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    11161 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    11221 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    11281 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    11341 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    11401 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    11461 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
    11521 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
    11581 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
    11641 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
    11701 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
    11761 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
    11821 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
    11881 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
    11941 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
    12001 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
    12061 tacttggcag tacatctacg tattagtcat cgctattacc atg
//
