LOCUS       sequence-181737-       12106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for SaCas9 knockout, lentiviral expression of S. aureus Cas9 
            (SauCas9) and gRNA scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12106)
  AUTHORS   Najm FJ, Strand C, Donovan KF, Hegde M, Sanson KR, Vaimberg EW, 
            Sullender ME, Hartenian E, Kalani Z, Fusi N, Listgarten J, Younger 
            ST, Bernstein BE, Root DE, Doench JG
  TITLE     Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens.
  JOURNAL   Nat Biotechnol. 2018 Feb;36(2):179-189. doi: 10.1038/nbt.4048. Epub 
            2017 Dec 18.
  PUBMED    29251726
REFERENCE   2  (bases 1 to 12106)
  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/nbt.4048"; 
            journalName: "Nat Biotechnol"; date: "2018-02"; volume: "36"; issue:
            "2"; pages: "179-189"
COMMENT     Sequence Label: pXPR_206
FEATURES             Location/Qualifiers
     source          1..12106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(112..132)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        117..446
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      297..432
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     359..378
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        494..541
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             560..934
                     /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    1064..1185
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1101..1120)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1155..1174
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1234..1250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1234..1250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1247..1269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1258..1274
                     /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(1282..1288,1289..1306,1307..1312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1327..1348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1465..1482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1636..2224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1716..1735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2395..3186,3187..3255))
                     /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     3018..3037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3256..3360)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3435..3454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3626..4005
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4007..4205
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4160..4180
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4223..4403
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4450..4575
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5068..5301
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5486..5530
                     /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    5828..5945
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5996..6244
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5996..6016
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6167..6186
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6269..6344
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     promoter        6406..6617
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6625..6634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6637..6657
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6682..9837
                     /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             9838..9885
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9895..9951
                     /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    10580..11168
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10633..10653)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11051..11062)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11246..11479
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11505..11522)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11511..11735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(11781..12106,1..103)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11868..11887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12078..12099
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
       61 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
      121 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      181 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
      241 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
      301 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
      361 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
      421 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
      481 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
      541 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
      601 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
      661 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
      721 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
      781 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
      841 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
      901 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
      961 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     1021 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     1081 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1141 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     1201 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     1261 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     1321 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     1381 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1441 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1501 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1561 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1621 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1681 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1741 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1801 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1861 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1921 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1981 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2041 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2101 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2161 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2221 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2281 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2341 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2401 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2461 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2521 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2581 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2641 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2701 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2761 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2821 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2881 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     2941 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3001 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3061 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3121 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3181 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3241 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3301 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3361 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     3421 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     3481 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     3541 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     3601 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     3661 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     3721 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     3781 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     3841 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     3901 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     3961 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     4021 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     4081 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     4141 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     4201 agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag cctgggagct
     4261 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     4321 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     4381 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     4441 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     4501 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     4561 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     4621 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     4681 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     4741 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     4801 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     4861 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     4921 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     4981 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     5041 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     5101 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     5161 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     5221 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     5281 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     5341 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     5401 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     5461 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     5521 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     5581 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     5641 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     5701 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     5761 tcgattagtg aacggatcgg cactgcgtgc gccaattctg cagacaaatg gcagtattca
     5821 tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     5881 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     5941 attttcgggt ttattacagg gacagcagag atccagtttg gttaattaag gtaccgaggg
     6001 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
     6061 tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
     6121 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct
     6181 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa
     6241 caccggagac gaacctagca tcgtctctgt ttaagtactc tggaaacaga atctacttaa
     6301 acaaggcaaa atgccgtgtt tatctcgtca acttgttggc gagatttttt gaattcgcta
     6361 gctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
     6421 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa
     6481 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     6541 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     6601 ttgccgccag aacacaggac cggtgccacc atggcaccca agaagaagcg caaagttggc
     6661 atccacggcg tgcctgctgc caagcgcaac tacatcctgg gcctggacat tggcatcacc
     6721 tcagttggct atggcatcat cgactacgag acccgggacg tcatcgacgc tggcgtccgc
     6781 ctcttcaagg aggccaacgt cgagaacaac gagggccggc ggtccaagcg cggggcccgc
     6841 cggctcaagc gccgccgccg ccaccgcatc cagcgcgtga aaaagttgct cttcgactac
     6901 aacttgctca ccgaccactc cgagctgtct ggcatcaacc cgtacgaggc ccgggtcaag
     6961 ggcctctcgc aaaagttgtc cgaggaggag ttctctgctg ccctgctgca cctggccaag
     7021 cgccgcgggg tccacaacgt caacgaagtt gaggaggaca ccggcaacga actttccacc
     7081 aaggagcaga tctcccgcaa ctccaaggcc ctggaggaga agtacgtcgc ggagctccaa
     7141 cttgagcgcc tcaagaagga cggcgaggtg cgcggctcca tcaaccgctt caagacctcc
     7201 gactacgtca aggaggccaa gcagctgctc aaggtccaga aggcctacca ccaacttgac
     7261 cagtccttca tcgacaccta catcgacctg ctggagaccc gccgcaccta ctacgagggg
     7321 cctggcgagg gctctccctt cggctggaag gacatcaagg agtggtacga gatgctcatg
     7381 ggccactgca cctacttccc ggaggagctg cgctcggtca agtacgccta caacgccgac
     7441 ctctacaacg cgctcaacga cctcaacaac ttggtcatca cccgcgacga gaacgaaaag
     7501 ttggagtact acgaaaagtt ccagatcatc gaaaacgtct tcaagcagaa gaagaagcca
     7561 actttgaagc agatcgccaa ggagatcctg gtcaacgagg aggacatcaa gggctaccgg
     7621 gtcacctcca ccggcaagcc cgagttcacc aacttaaagg tctaccatga catcaaggac
     7681 atcaccgccc gcaaggagat catcgagaac gcggagctgc tggaccagat cgccaagatc
     7741 ctcaccatct accaaagttc cgaggacatc caggaggagc tgaccaactt aaactcggag
     7801 cttacccagg aggagatcga gcagatctcc aacttaaagg gctacactgg cacccacaac
     7861 ttgtccctca aggccatcaa cttgatcctg gacgaacttt ggcataccaa cgacaaccag
     7921 atcgccatct tcaaccgact aaagttggtg cccaagaaag ttgacctgtc ccaacagaag
     7981 gagatcccaa ctaccctggt cgacgacttc atcctctcgc ccgtggtcaa gcgcagcttc
     8041 atccagtcca tcaaggtcat caacgccatc atcaagaagt acggcctgcc caacgacatc
     8101 atcatcgagt tggcacgcga gaagaactcc aaggatgccc agaagatgat caacgagatg
     8161 cagaagcgca accgccagac caacgagcgc atcgaggaga tcatccgcac caccggcaag
     8221 gagaacgcca agtacctgat cgagaagata aagttgcatg acatgcagga gggcaagtgc
     8281 ctctactccc tggaggccat ccccctcgag gacctgctca acaacccctt caactacgaa
     8341 gttgaccaca tcatcccgcg ctcggtgtcc ttcgacaaca gcttcaacaa caaggtcctg
     8401 gtcaagcagg aggagaactc caagaagggc aaccggaccc ccttccagta cctgtcctcc
     8461 tcggactcca agatctccta cgaaactttc aagaagcaca tcctcaactt ggcaaagggc
     8521 aagggccgca tctccaagac caagaaggag tacctgctgg aggagcgcga catcaaccgc
     8581 ttctcggtgc agaaggactt catcaaccgc aacctggtgg acacccgcta cgcaacccgt
     8641 ggcctcatga acctgctgcg cagctacttc cgggtcaaca acttggacgt caaagttaag
     8701 tcgatcaacg gtggcttcac cagcttcctg cgccgcaagt ggaagttcaa gaaggagcgc
     8761 aacaagggct acaagcacca tgcggaggac gccctcatca tcgccaacgc cgacttcatc
     8821 ttcaaggagt ggaaaaagtt ggacaaggcc aagaaggtca tggagaacca gatgttcgag
     8881 gagaagcagg ccgagtccat gccggagatc gagacagagc aggagtacaa ggagatcttc
     8941 atcacgccgc accagatcaa gcacatcaag gacttcaagg actacaagta ctcccaccgc
     9001 gtggacaaga agcccaaccg cgagctcatc aacgacaccc tctactccac ccgcaaggac
     9061 gacaagggca acaccctcat cgtcaacaac ttaaacggcc tctacgacaa ggacaacgac
     9121 aagttgaaaa agttgatcaa caagagcccc gaaaagttgc tcatgtacca ccatgacccg
     9181 cagacgtacc aaaagttgaa gttgatcatg gagcagtacg gtgacgagaa gaacccgctc
     9241 tacaagtact acgaggagac cggcaactac ctcaccaagt actccaagaa ggacaacggc
     9301 ccggtcatca agaagatcaa gtactacggc aacaagttga atgcccacct ggacatcacc
     9361 gacgactacc ccaactcccg caacaaagtt gtaaagttgt cgctaaagcc gtaccgcttc
     9421 gacgtctacc tggacaacgg cgtctacaag ttcgtgaccg tcaagaacct ggacgtcatc
     9481 aagaaggaga actactacga ggtcaactcc aagtgctacg aggaggccaa aaagttgaag
     9541 aagatctcca accaggccga gttcatcgcc agcttctaca acaacgacct catcaagatc
     9601 aacggcgagc tctaccgcgt catcggggtc aacaacgacc tgctgaaccg catcgaggtc
     9661 aacatgatcg acatcaccta ccgcgagtac ctcgagaaca tgaacgacaa gcgcccgcct
     9721 cgcatcatca agaccatcgc ctccaagacc cagtccatca agaagtactc caccgacatc
     9781 cttggcaact tgtacgaggt caagagcaag aagcaccctc agatcatcaa gaagggcaag
     9841 cgccctgctg ccaccaagaa ggctggccag gccaagaaga agaagggatc cggggccacc
     9901 aacttttccc tgctgaagca ggctggcgac gtcgaggaga accctggccc tggctccgga
     9961 accgagtaca agccaactgt ccgcttggca acccgcgacg atgtgcccag ggccgtccgc
    10021 acgctggctg ctgccttcgc cgactaccca gccacccgcc acaccgtgga cccggaccgc
    10081 cacatcgagc gcgtcaccga gctgcaggag ctcttcctca cccgagttgg ccttgacatt
    10141 ggcaaggtct gggtggcgga cgacggtgct gccgtggccg tctggaccac ccccgagtcg
    10201 gtcgaggctg gcgccgtctt cgccgagatc gggccgcgca tggccgagct ttcgggctcc
    10261 cggttggctg cccaacagca gatggagggg ctgttggcac cccaccgccc caaggagccg
    10321 gcctggttcc tggccacagt tggtgtctct cccgaccacc agggcaaggg cctgggcagc
    10381 gccgtggtgc tgcctggggt cgaggctgcc gagcgcgctg gcgtccctgc cttcctggag
    10441 acctctgccc ctcgcaactt gcccttctac gagcgcctgg gcttcaccgt caccgcggac
    10501 gtcgaggtgc ccgaggggcc gcgcacctgg tgcatgaccc gcaagcctgg ggcatgaatc
    10561 tagaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
    10621 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
    10681 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
    10741 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
    10801 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
    10861 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
    10921 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
    10981 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
    11041 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
    11101 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
    11161 ctccccgcag cgctgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc
    11221 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat
    11281 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
    11341 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
    11401 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
    11461 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
    11521 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
    11581 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
    11641 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
    11701 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
    11761 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
    11821 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
    11881 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
    11941 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
    12001 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
    12061 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaaca
//
