LOCUS       sequence-139517-        8388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  optimized lenti sgRNA cloning backbone with MS2 loops at tetraloop 
            and stemloop 2 and EF1a-puro resistance marker. Contains BsmBI sites
            for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8388)
  AUTHORS   Joung J, Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham
            MD, Sanjana NE, Zhang F
  TITLE     Genome-scale CRISPR-Cas9 knockout and transcriptional activation 
            screening.
  JOURNAL   Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. 
            Epub 2017 Mar 23.
  PUBMED    28333914
REFERENCE   2  (bases 1 to 8388)
  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: 
            "10.1038/nprot.2017.016"; journalName: "Nat Protoc"; date: 
            "2017-04"; volume: "12"; issue: "4"; pages: "828-863"
COMMENT     Sequence Label: lenti sgRNA(MS2)_puro optimized backbone
FEATURES             Location/Qualifiers
     source          1..8388
                     /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        287..305
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        357..375
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    467..584
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        723..1901
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          954..1892
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1849..1869
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1914..2513
                     /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(1914..1933)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2410..2430
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2529..3117
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2582..2602)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3000..3011)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3189..3422
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3500..3621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3537..3556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3591..3610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3661..3796
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3816..3835)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3817..3835)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3845..3862)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3845..3861)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3854..3876)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4003..4458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4090..4109)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4300..4321
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4484..4588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4589..4657,4658..5449)
                     /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(4807..4826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5620..6208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6109..6128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6362..6379
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6496..6517
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6532..6537,6538..6555,6556..6562)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6570..6586
                     /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     6575..6597
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6594..6610
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6594..6610
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6629..6649
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6631..6649
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6677..6903
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6904..7084
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7131..7256
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7749..7982
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8167..8211
                     /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 cgaaacaccc gagacgggat accgtctctg ttttagagct aggccaacat gaggatcacc
      301 catgtctgca gggcctagca agttaaaata aggctagtcc gttatcaact tggccaacat
      361 gaggatcacc catgtctgca gggccaagtg gcaccgagtc ggtgcttttt tggatccaag
      421 cttggcgtaa ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
      481 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
      541 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
      601 acagcagaga tccactttgg cgccggctcg agggggcccg ggtgcaaaga tggataaagt
      661 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      721 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      781 ggggaggggt cggcaattga tccggtgcct agagaaggtg gcgcggggta aactgggaaa
      841 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      901 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      961 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     1021 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     1081 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     1141 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     1201 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     1261 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     1321 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     1381 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     1441 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     1501 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     1561 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1621 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1681 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1741 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1801 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1861 cagtggttca aagttttttt cttccatttc aggtgtcgtg acgtacggcc accatgaccg
     1921 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     1981 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     2041 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     2101 aggtgtgggt cgcggacgac ggcgccgccg tggcggtctg gaccacgccg gagagcgtcg
     2161 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     2221 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     2281 ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     2341 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     2401 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     2461 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaacgcgtt
     2521 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2581 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2641 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2701 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2761 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2821 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2881 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     2941 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3001 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3061 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
     3121 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     3181 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
     3241 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     3301 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     3361 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     3421 cagtacgtat agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg
     3481 aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa
     3541 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3601 caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc
     3661 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3721 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3781 ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc
     3841 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     3901 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     3961 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     4021 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4081 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4141 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4201 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4261 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4321 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4381 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4441 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4501 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4561 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4621 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4681 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4741 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4801 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4861 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4921 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4981 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5041 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5101 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5161 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5221 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5281 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5341 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5401 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5461 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5521 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5581 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5641 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5701 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5761 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5821 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5881 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5941 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6001 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6061 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6121 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6181 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6241 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6301 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6361 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6421 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6481 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6541 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6601 cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct
     6661 ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg
     6721 atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa
     6781 gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga
     6841 cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata
     6901 aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6961 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     7021 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     7081 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
     7141 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
     7201 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
     7261 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     7321 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     7381 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     7441 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     7501 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     7561 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     7621 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     7681 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     7741 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     7801 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     7861 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     7921 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     7981 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     8041 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     8101 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     8161 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     8221 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     8281 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     8341 tcaccattat cgtttcagac ccacctccca accccgaggg gacccaga
//
