LOCUS       sequence-350670-        8323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing PolB-gRNA-1 and a puromycin resistance 
            cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8323)
  AUTHORS   Fang Q, Andrews J, Sharma N, Wilk A, Clark J, Slyskova J, Koczor CA,
            Lans H, Prakash A, Sobol RW
  TITLE     Stability and sub-cellular localization of DNA polymerase beta is 
            regulated by interactions with NQO1 and XRCC1 in response to 
            oxidative stress.
  JOURNAL   Nucleic Acids Res. 2019 Jul 9;47(12):6269-6286. doi: 
            10.1093/nar/gkz293.
  PUBMED    31287140
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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2019-07-9- 9"; volume: "47"; issue: 
            "12"; pages: "6269-6286"
COMMENT     Sequence Label: pLentiGuide-PolB1-puro
FEATURES             Location/Qualifiers
     source          1..8323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        152..1330
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          383..1321
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1278..1298
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1343..1942
                     /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(1343..1362)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1839..1859
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    1958..2546
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2011..2031)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2429..2440)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2618..2851
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2929..3050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2966..2985)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3020..3039
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3090..3225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3245..3264)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3246..3264)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3274..3291)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3274..3290)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3283..3305)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3432..3887
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3519..3538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3729..3750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3913..4017
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4018..4086,4087..4878)
                     /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(4236..4255)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5049..5637
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5538..5557
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5791..5808
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5925..5946
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5961..5966,5967..5984,5985..5991)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5999..6015
                     /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     6004..6026
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6023..6039
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6023..6039
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6058..6078
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6060..6078
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6106..6332
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6333..6513
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6560..6685
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7178..7411
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7596..7640
                     /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        7818..8066
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7818..7838
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7989..8008
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8088..8163
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    join(8219..8323,1..13)
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga
       61 gggggcccgg gtgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg
      121 gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
      181 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta
      241 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
      301 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
      361 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
      421 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct
      481 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
      541 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat
      601 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
      661 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
      721 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
      781 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
      841 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
      901 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
      961 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     1021 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     1081 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     1141 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     1201 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     1261 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     1321 ggtgtcgtga cgtacggcca ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     1381 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     1441 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     1501 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgccgt
     1561 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     1621 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     1681 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga
     1741 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     1801 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     1861 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     1921 gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat
     1981 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     2041 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     2101 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     2161 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     2221 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     2281 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     2341 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     2401 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     2461 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     2521 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     2581 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     2641 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     2701 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     2761 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     2821 cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt
     2881 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     2941 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3001 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     3061 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3121 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3181 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     3241 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     3301 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     3361 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     3421 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3481 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3541 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3601 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3661 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3721 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3781 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3841 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     3901 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3961 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4021 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4081 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4141 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4201 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4261 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4321 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4381 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4441 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4501 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4561 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4621 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4681 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4741 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4801 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4861 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4921 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4981 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5041 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5101 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5161 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5221 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5281 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5341 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5401 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5461 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5521 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5581 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5641 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5701 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5761 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5821 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     5881 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     5941 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6001 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     6061 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     6121 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     6181 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     6241 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     6301 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
     6361 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6421 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6481 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     6541 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     6601 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     6661 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     6721 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     6781 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     6841 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     6901 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     6961 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     7021 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     7081 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     7141 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     7201 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     7261 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     7321 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     7381 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     7441 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     7501 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     7561 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     7621 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     7681 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     7741 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     7801 ccccgagggg acccagagag ggcctatttc ccatgattcc ttcatatttg catatacgat
     7861 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac
     7921 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     7981 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     8041 atatcttgtg gaaaggacga aacaccggag caaacggaag gcgccgcgtt ttagagctag
     8101 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     8161 tgctttttta agcttggcgt aactagatct tgagacaaat ggcagtattc atccacaatt
     8221 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     8281 caacagacat acaaactaaa gaattacaaa aacaaattac aaa
//
