LOCUS       sequence-169870-       10952 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of guide RNA (mup53_Ex4) with fluorescent GFP 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10952)
  AUTHORS   Aubrey BJ, Kelly GL, Kueh AJ, Brennan MS, O'Connor L, Milla L, 
            Wilcox S, Tai L, Strasser A, Herold MJ
  TITLE     An inducible lentiviral guide RNA platform enables the 
            identification of tumor-essential genes and tumor-promoting 
            mutations in vivo.
  JOURNAL   Cell Rep. 2015 Mar 3;10(8):1422-32. doi: 
            10.1016/j.celrep.2015.02.002. Epub 2015 Feb 26.
  PUBMED    25732831
REFERENCE   2  (bases 1 to 10952)
  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: 
            "10.1016/j.celrep.2015.02.002"; journalName: "Cell Rep"; date: 
            "2015-03-3- 3"; volume: "10"; issue: "8"; pages: "1422-32"
COMMENT     Sequence Label: FgH1tUTG_mup53_Ex4
FEATURES             Location/Qualifiers
     source          1..10952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..32
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(151..171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        156..485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      336..471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     398..417
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        533..580
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             599..973
                     /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    1103..1224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1140..1159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1194..1213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1273..1289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1273..1289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1286..1308)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1297..1313
                     /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(1321..1327,1328..1345,1346..1351))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1366..1387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1504..1521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1675..2263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1755..1774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2434..3225,3226..3294))
                     /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     3057..3076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3295..3399)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3474..3493)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3665..4044
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4046..4244
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4199..4219
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4262..4442
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4489..4614
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5107..5340
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5525..5569
                     /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    5867..5984
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     6175..6194
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    6239..6257
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_RNA        6283..6358
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6394..7605
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     7442..7461
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             7643..8263
                     /codon_start=1
                     /gene="synthetic gene"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     CDS             8273..8326
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(8327..8329,8330..8332,8333..9046)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(8327..8329,8330..8332,8333..9043)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8372..8393)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8633..8652)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8980..9001
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    9086..9674
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9139..9159)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9557..9568)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10184..10364
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10390..10407)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10396..10620
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(10666..10952,1..142)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10753..10772)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
       61 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      121 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
      181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      421 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      481 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
      541 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
      601 ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga
      661 gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt
      721 ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa
      781 caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt
      841 cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc
      901 gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga
      961 ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
     1021 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     1081 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     1141 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1201 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     1261 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1321 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1381 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1441 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1501 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1561 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1621 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1681 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1741 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1801 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1861 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1921 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1981 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2041 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2101 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2161 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2221 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2281 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2341 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2401 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2461 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2521 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2581 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2641 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2701 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2761 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2821 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2881 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2941 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3001 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3061 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3121 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3181 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3241 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3301 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3361 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3421 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     3481 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     3541 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     3601 aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg
     3661 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     3721 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     3781 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     3841 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3901 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3961 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     4021 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     4081 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     4141 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     4201 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac
     4261 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     4321 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     4381 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     4441 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     4501 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     4561 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     4621 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     4681 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     4741 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     4801 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     4861 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     4921 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     4981 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     5041 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     5101 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     5161 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     5221 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     5281 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     5341 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     5401 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     5461 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     5521 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     5581 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     5641 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     5701 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     5761 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc
     5821 actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     5881 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     5941 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     6001 acagcagaga tccagtttgg ttaattaacc atggaattcg aacgctgacg tcatcaaccc
     6061 gctccaagga atcgcgggcc cagtgtcact aggcgggaac acccagcgcg cgtgcgccct
     6121 ggcaggaaga tggctgtgag ggacagggga gtggcgccct gcaatatttg catgtcgcta
     6181 tgtgttctgg gaaatcacca taaacgtgaa atgtctttgg atttgggaat cttataagtc
     6241 cctatcagtg atagagattc ccggcaacta tggcttccac ctgttttaga gctagaaata
     6301 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     6361 tttttttaat taacccgtgt cggctccaga tctggcctcc gcgccgggtt ttggcgcctc
     6421 ccgcgggcgc ccccctcctc acggcgagcg ctgccacgtc agacgaaggg cgcagcgagc
     6481 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     6541 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     6601 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     6661 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     6721 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     6781 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     6841 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     6901 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     6961 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     7021 gtgggcggca agaacccaag gtcttgaggc cttcgctaat gcgggaaagc tcttattcgg
     7081 gtgagatggg ctggggcacc atctggggac cctgacgtga agtttgtcac tgactggaga
     7141 actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc
     7201 cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat
     7261 aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg
     7321 cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg
     7381 gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag
     7441 ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt
     7501 taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagactagt
     7561 aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacgaagc ttgggctgca
     7621 ggtcgactct agaggatcca ccatgtccag actggacaag agcaaagtca tcaactctgc
     7681 cttggagctc ctgaatgaag ttggcattga gggcttgacc accaggaagc tggcccagaa
     7741 gctgggtgtg gagcagccta ccctgtactg gcatgtgaag aacaagaggg ctctgcttga
     7801 tgccctggcc attgagatgt tggacaggca ccacacccac ttctgccctc tggaagggga
     7861 gtcctggcag gacttcctga ggaacaatgc caagagcttc agatgtgcct tgctctccca
     7921 ccgggatggt gccaaagttc acttgggcac caggcctaca gagaagcagt atgagaccct
     7981 ggagaaccag ctggcattcc tgtgccaaca aggcttctcc ctggaaaatg ccttgtatgc
     8041 cctctctgct gtgggccact tcaccttggg ctgtgtgctg gaggaccagg agcaccaagt
     8101 tgccaaggag gagagggaga cccccaccac tgactccatg ccaccactgc tgcggcaagc
     8161 tattgagttg tttgaccacc aaggggctga gcctgcattc ctttttggcc tggaactgat
     8221 catctgtggc ctggaaaagc agctgaaatg tgagtctggc tctgggtccg gtgagggcag
     8281 aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccaatgg tgagcaaggg
     8341 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     8401 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     8461 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     8521 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     8581 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     8641 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     8701 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     8761 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     8821 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     8881 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     8941 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     9001 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgatct
     9061 acaattcgat atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     9121 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     9181 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     9241 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     9301 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     9361 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     9421 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     9481 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     9541 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     9601 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     9661 ggccgcctcc ccgcatcgag acctagaaaa acatggagca atcacaagta gcaatacagc
     9721 agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg tgggttttcc
     9781 agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca
     9841 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatat
     9901 ccttgatctg tggatctacc acacacaagg ctacttccct gattggcaga actacacacc
     9961 agggccaggg atcagatatc cactgacctt tggatggtgc tacaagctag taccagttga
    10021 gcaagagaag gtagaagaag ccaatgaagg agagaacacc cgcttgttac accctgtgag
    10081 cctgcatggg atggatgacc cggagagaga agtattagag tggaggtttg acagccgcct
    10141 agcatttcat cacatggccc gagagctgca tccggactgt actgggtctc tctggttaga
    10201 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
    10261 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
    10321 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc
    10381 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
    10441 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
    10501 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
    10561 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
    10621 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
    10681 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
    10741 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
    10801 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
    10861 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
    10921 cggtttttcg ccctttgacg ttggagtcca cg
//
