LOCUS       sequence-231639-        8323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8323)
  AUTHORS   Li J, Choi PS, Chaffer CL, Labella K, Hwang JH, Giacomelli AO, Kim 
            JW, Ilic N, Doench JG, Ly SH, Dai C, Hagel K, Hong AL, Gjoerup O, 
            Goel S, Ge JY, Root DE, Zhao JJ, Brooks AN, Weinberg RA, Hahn WC
  TITLE     An alternative splicing switch in FLNB promotes the mesenchymal cell
            state in human breast cancer.
  JOURNAL   Elife. 2018 Jul 30;7. pii: 37184. doi: 10.7554/eLife.37184.
  PUBMED    30059005
REFERENCE   2  (bases 1 to 8323)
  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: "Elife."; 
            date: "2018-07-30"; volume: "7. pii: 37184. doi"; pages: " 
            10.7554/eLife.37184"
COMMENT     Sequence Label: pXPR_sgFLNB-SK2
FEATURES             Location/Qualifiers
     source          1..8323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    402..519
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        658..1836
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          889..1827
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1784..1804
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1849..2448
                     /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(1849..1868)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2345..2365
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2464..3052
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2517..2537)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2935..2946)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3124..3357
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3435..3556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3472..3491)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3526..3545
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3596..3731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3751..3770)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3752..3770)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3780..3797)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3780..3796)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3789..3811)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3938..4393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4025..4044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4235..4256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4419..4523
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4524..4592,4593..5384)
                     /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(4742..4761)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5555..6143
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6044..6063
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6297..6314
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6431..6452
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6467..6472,6473..6490,6491..6497)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6505..6521
                     /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     6510..6532
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6529..6545
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6529..6545
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6564..6584
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6566..6584
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6612..6838
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6839..7019
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7066..7191
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7684..7917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8102..8146
                     /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"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ccactgacca ggccacagat gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttaagcttgg
      361 cgtaactaga tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg
      421 attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact
      481 aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc
      541 agagatccac tttggcgccg gctcgagggg gcccgggtgc aaagatggat aaagttttaa
      601 acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt gggaattggc
      661 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
      721 ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
      781 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
      841 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
      901 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
      961 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
     1021 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     1081 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     1141 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     1201 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     1261 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     1321 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
     1381 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     1441 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     1501 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     1561 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     1621 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     1681 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     1741 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     1801 gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta cggccaccat gaccgagtac
     1861 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     1921 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     1981 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     2041 tgggtcgcgg acgacggcgc cgccgtggcg gtctggacca cgccggagag cgtcgaagcg
     2101 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     2161 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     2221 ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     2281 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     2341 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     2401 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaac gcgttaagtc
     2461 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2521 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2581 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2641 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2701 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2761 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2821 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     2881 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     2941 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3001 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcgtcgactt
     3061 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     3121 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
     3181 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     3241 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     3301 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
     3361 cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     3421 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3481 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3541 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc
     3601 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3661 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3721 ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     3781 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     3841 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     3901 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     3961 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4021 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4081 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4141 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4201 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4261 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4321 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4381 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4441 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4501 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4561 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4621 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4681 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4741 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4801 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4861 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4921 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4981 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5041 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     5101 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5161 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5221 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5281 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5341 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5401 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5461 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5521 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5581 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5641 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5701 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5761 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5821 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5881 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5941 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6001 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6061 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6121 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     6181 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6241 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6301 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     6361 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     6421 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     6481 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     6541 atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc
     6601 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag
     6661 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag
     6721 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac
     6781 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg
     6841 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     6901 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     6961 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     7021 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
     7081 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
     7141 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
     7201 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
     7261 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
     7321 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
     7381 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
     7441 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
     7501 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
     7561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     7621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     7681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     7741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     7801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     7861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     7921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     7981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     8041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     8101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     8161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     8221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     8281 attatcgttt cagacccacc tcccaacccc gaggggaccc aga
//
