LOCUS       pRC0162-V2_pSB700-Puromycin        8941 bp ds-DNA     circular SYN 31-JAN-2022
DEFINITION  Lentiviral vector for expressing U6 sgRNA and CAGGS Cerulean
            fluorescent protein.
ACCESSION
VERSION     .
KEYWORDS    pSB700.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct.
REFERENCE   1  (bases 1 to 9061)
  AUTHORS   Chavez, A, Scheiman, J, Vora, S, Pruitt, BW, Tuttle, M, P, R, Iyer,
            E, Lin, S, Kiani, S, Guzman, CD, Wiegand, DJ, Ter-Ovanesyan, D,
            Braff, JL, Davidsohn, N, Housden, BE, Perrimon, N, Weiss, R, Aach,
            J, Collins, JJ, Church and GM
  TITLE     Highly efficient Cas9-mediated transcriptional programming.
  JOURNAL   Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312.
  PUBMED    25730490
REFERENCE   2  (bases 1 to 9061)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 20, 2017 from SnapGene Console 4.0.0a8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     source          1..8941
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /label="HIV-1 Psi"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to
                     thecytoplasm."
                     /label="RRE"
     misc_feature    2028..2143
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     PCR_primer      2209..2228
                     /created_by="charir2"
                     /label="IA"
     promoter        2246..2486
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /label="U6 promoter"
     primer_bind     2246..2266
                     /note="Human U6 promoter, forward primer"
                     /label="hU6-F"
     primer_bind     2417..2436
                     /note="Human U6 promoter, forward primer"
                     /label="LKO.1 5'"
     misc_RNA        2516..2591
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /label="g RNA scaffold"
     PCR_primer      complement(2599..2620)
                     /created_by="charir2"
                     /modified_by="charir2"
                     /label="R"
     enhancer        2665..2968
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2970..3244
                     /label="chicken beta-actin promoter"
     intron          3246..4263
                     /note="chimera between introns from chicken beta-actin and
                      rabbit beta-globin"
                     /label="chimeric intron"
     primer_bind     4187..4209
                     /note="Chimeric intron in CAG promoter, forward primer"
                     /label="pCAGGS-5"
     primer_bind     4271..4290
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
                     /label="pCAG-F"
     source          <4297..>4299
                     /dnas_title="1-0328-16_PB-Dox-DHFR-N-dCas9Sp-KRAB-P2A-mChe
                     rry"
     misc_feature    4297..>4299
                     /note="Puro"
     CDS             4300..4896
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLE
                     TSAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
                     /label="PuroR"
     source          <4300..>4896
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     4793..4813
                     /note="Puromycin resistance gene, forward primer"
                     /label="Puro-F"
     misc_feature    4909..5497
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /label="WPRE"
     primer_bind     complement(4962..4982)
                     /note="WPRE, reverse primer"
                     /label="WPRE-R"
     CDS             complement(5380..5391)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
                     /label="Factor Xa site"
     LTR             5704..6337
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6466..6482)
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /label="M13 rev"
     primer_bind     complement(6466..6482)
                     /note="In lacZ gene. Also called M13-rev"
                     /label="M13 Reverse"
     primer_bind     complement(6479..6501)
                     /note="In lacZ gene"
                     /label="M13/pUC Reverse"
     protein_bind    6490..6506
                     /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)."
                     /label="lac operator"
     promoter        complement(6514..6544)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6559..6580
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                      of cAMP."
                     /label="CAP binding site"
     primer_bind     complement(6697..6714)
                     /note="L4440 vector, forward primer"
                     /label="L4440"
     rep_origin      complement(6868..7456)
                     /direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori"
     primer_bind     complement(6948..6967)
                     /note="pBR322 origin, forward primer"
                     /label="pBR322ori-F"
     CDS             complement(7627..8487)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="AmpR"
     primer_bind     8250..8269
                     /note="Ampicillin resistance gene, reverse primer"
                     /label="Amp-R"
     promoter        complement(8488..8592)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8640..8774
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(8677..8696)
                     /note="SV40 polyA, reverse primer"
                     /label="SV40pA-R"
     primer_bind     8731..8750
                     /note="SV40 polyA terminator, reverse primer"
                     /label="EBV-rev"
ORIGIN      
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cattagtgaa cggatctcga cggtatcgat cacgagacta
     2221 gcctcgagcg gccgccccct tcaccgaggg cctatttccc atgattcctt catatttgca
     2281 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
     2341 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
     2401 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
     2461 ggctttatat atcttgtgga aaggacgaaa caccggagac gattaatgcg tctcggtttt
     2521 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     2581 cgagtcggtg ctttttttga attctcgacc tcgagacaaa tggcagtatt cgtcattagt
     2641 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     2701 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     2761 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     2821 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     2881 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     2941 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     3001 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     3061 tgcagcgatg ggggcggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg
     3121 gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa
     3181 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     3241 ggcggggagt cgctgcgacg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
     3301 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
     3361 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
     3421 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
     3481 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
     3541 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
     3601 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
     3661 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
     3721 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
     3781 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
     3841 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
     3901 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
     3961 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
     4021 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     4081 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     4141 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     4201 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
     4261 cagctcctgg gcaacgtgct ggttattgtg gccaccatga ccgagtacaa gcccacggtg
     4321 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     4381 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     4441 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     4501 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     4561 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     4621 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     4681 ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     4741 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     4801 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     4861 cgcacctggt gcatgacccg caagcccggt gcctgaagcg tctggaacaa tcaacctctg
     4921 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     4981 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5041 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     5101 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5161 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5221 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5281 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     5341 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     5401 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     5461 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag
     5521 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     5581 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     5641 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg
     5701 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     5761 acacacaagg ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc
     5821 cactgacctt tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg
     5881 ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc
     5941 cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc
     6001 gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt
     6061 ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca
     6121 gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc
     6181 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     6241 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     6301 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     6361 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc
     6421 tagcgtttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     6481 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     6541 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6601 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6661 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6721 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6781 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6841 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6901 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6961 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7021 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7081 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7141 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7201 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7261 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7321 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7381 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7441 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7501 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7561 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7621 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7681 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7741 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7801 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7861 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7921 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7981 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8041 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8101 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8161 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8221 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8281 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8341 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8401 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8461 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8521 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8581 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca
     8641 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     8701 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     8761 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
     8821 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct
     8881 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     8941 t
//
