LOCUS       sequence-325298-        9180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3rd generation lentivirus sgRNA delivery backbone with Cre-removable
            mKate2 reporter and puromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9180)
  AUTHORS   Dubrot J, Lane-Reticker SK, Kessler EA, Ayer A, Mishra G, Wolfe CH, 
            Zimmer MD, Du PP, Mahapatra A, Ockerman KM, Davis TGR, Kohnle IC, 
            Pope HW, Allen PM, Olander KE, Iracheta-Vellve A, Doench JG, Haining
            WN, Yates KB, Manguso RT
  TITLE     In vivo screens using a selective CRISPR antigen removal lentiviral 
            vector system reveal immune dependencies in renal cell carcinoma.
  JOURNAL   Immunity. 2021 Jan 20. pii: S1074-7613(21)00001-7. doi: 
            10.1016/j.immuni.2021.01.001.
  PUBMED    33497609
REFERENCE   2  (bases 1 to 9180)
  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: "Immunity. 
            2021 Jan 20. pii: S1074-7613(21)00001-7. doi: 
            10.1016/j.immuni.2021.01.001."
COMMENT     Sequence Label: pSCAR_sgRNA_puro-mKate-lox5171
FEATURES             Location/Qualifiers
     source          1..9180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      711..1299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1200..1219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1453..1470
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1587..1608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1623..1628,1629..1646,1647..1653)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1661..1677
                     /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)."
     primer_bind     1666..1688
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1685..1701
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1685..1701
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1720..1740
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1722..1740
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1768..1994
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1995..2175
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2222..2347
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2840..3073
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3258..3302
                     /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"
     promoter        3483..3731
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3483..3503
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3654..3673
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3753..3828
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3890..4007
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(4066..4099)
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        4180..5358
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4411..5349
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5306..5326
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5371..5967
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5371..5390)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5867..5887
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             5989..6045
                     /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             6052..6750
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     protein_bind    complement(6751..6784)
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     misc_feature    6800..7388
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6853..6873)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7271..7282)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7460..7693
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7771..7892
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7808..7827)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7862..7881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7932..8067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8087..8106)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8088..8106)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8116..8133)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8116..8132)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8125..8147)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8274..8729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8361..8380)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8571..8592
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8755..8859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8860..8928,8929..9180,1..540)
                     /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     complement(9078..9097)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
       61 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
      121 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
      181 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
      241 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
      301 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
      361 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
      421 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
      481 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
      541 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
      601 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
      661 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
      721 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
      781 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
      841 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
      901 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
      961 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1021 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     1081 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     1141 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     1201 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     1261 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     1321 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     1381 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     1441 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     1501 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     1561 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     1621 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     1681 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     1741 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     1801 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     1861 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     1921 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     1981 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     2041 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     2101 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     2161 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
     2221 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
     2281 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
     2341 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
     2401 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     2461 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     2521 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     2581 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     2641 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     2701 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     2761 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     2821 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     2881 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     2941 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     3001 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     3061 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     3121 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     3181 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     3241 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     3301 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     3361 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     3421 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccggta
     3481 ccgagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
     3541 agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
     3601 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
     3661 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
     3721 gacgaaacac cggagacgct tttttcgtct cagtttgaga gctagaaata gcaagttcaa
     3781 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgaattc
     3841 aagcttggcg taactagatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa
     3901 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     3961 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     4021 gggacagcag agatccactt tggcgccggc tcgagggggc ccgggataac ttcgtatagt
     4081 acacattata cgaagttatt gcaaagatgg ataaagtttt aaacagagag gaatctttgc
     4141 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     4201 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgatcc
     4261 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     4321 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     4381 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
     4441 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
     4501 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     4561 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
     4621 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     4681 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     4741 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     4801 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     4861 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     4921 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     4981 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     5041 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     5101 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     5161 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     5221 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     5281 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     5341 ccatttcagg tgtcgtgacg tacggccacc atgaccgagt acaagcccac ggtgcgcctc
     5401 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5461 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     5521 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5581 gccgccgtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5641 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5701 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtt
     5761 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5821 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5881 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5941 tggtgcatga cccgcaagcc cggtgccgct agcctgcagg gatccggcgc aacaaacttc
     6001 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccggctag catggtgagc
     6061 gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt gaacaaccac
     6121 cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca gaccatgaga
     6181 atcaaggcgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc taccagcttc
     6241 atgtacggca gcaaaacctt catcaaccac acccagggca tccccgactt ctttaagcag
     6301 tccttccccg agggcttcac atgggagaga gtcaccacat acgaagacgg gggcgtgctg
     6361 accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt caagatcaga
     6421 ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg ctgggaggcc
     6481 tccaccgaga ccctgtaccc cgctgacggc ggcctggaag gcagagccga catggccctg
     6541 aagctcgtgg gcgggggcca cctgatctgc aacttgaaga ccacatacag atccaagaaa
     6601 cccgctaaga acctcaagat gcccggcgtc tactatgtgg acagaagact ggaaagaatc
     6661 aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc cagatactgc
     6721 gacctcccta gcaaactggg gcacagataa ataacttcgt atagtacaca ttatacgaag
     6781 ttatacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     6841 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     6901 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     6961 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     7021 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     7081 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     7141 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     7201 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     7261 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     7321 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     7381 ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     7441 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     7501 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     7561 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7621 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7681 aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt
     7741 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     7801 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     7861 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     7921 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     7981 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     8041 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     8101 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     8161 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     8221 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     8281 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     8341 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     8401 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     8461 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     8521 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     8581 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     8641 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     8701 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     8761 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     8821 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     8881 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     8941 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     9001 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     9061 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     9121 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
//
