LOCUS       sequence-379893-        8388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of sgRNA targeting mHbb-bh1promoter to 
            activate mouse Hbb-bh1 transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8388)
  AUTHORS   Omachi K, Miner JH
  TITLE     Comparative analysis of dCas9-VP64 variants and multiplexed guide 
            RNAs mediating CRISPR activation.
  JOURNAL   PLoS One. 2022 Jun 28;17(6):e0270008. doi: 
            10.1371/journal.pone.0270008. eCollection 2022.
  PUBMED    35763517
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"; journalName: "PLoS One.";
            date: "2022-06-28"; volume: "17(6):e0270008. doi"; pages: " 
            10.1371/journal.pone.0270008. eCollection 2022"
COMMENT     Sequence Label: LentiU6-mHbb-bh1_ gRNA_1-MS2-Puro
FEATURES             Location/Qualifiers
     source          1..8388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        162..410
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     162..182
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     333..352
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        449..467
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        519..537
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    628..745
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        884..2062
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1115..2053
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2010..2030
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2075..2674
                     /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(2075..2094)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2571..2591
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2690..3278
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2743..2763)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3161..3172)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3350..3583
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3661..3782
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3698..3717)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3752..3771
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3822..3957
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3977..3996)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3978..3996)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4006..4023)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4006..4022)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4015..4037)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4164..4619
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4251..4270)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4461..4482
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4645..4749
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4750..4818,4819..5610)
                     /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(4968..4987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5781..6369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6270..6289
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6523..6540
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6657..6678
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6693..6698,6699..6716,6717..6723)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6731..6747
                     /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     6736..6758
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6755..6771
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6755..6771
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6790..6810
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6792..6810
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6838..7064
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7065..7245
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7292..7417
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7910..8143
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8328..8372
                     /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 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
       61 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
      121 tatcgtttca gacccacctc ccaaccccga ggggacccag agagggccta tttcccatga
      181 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga
      241 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
      301 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
      361 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc gagagagtct
      421 gggcaagaca ggttttagag ctaggccaac atgaggatca cccatgtctg cagggcctag
      481 caagttaaaa taaggctagt ccgttatcaa cttggccaac atgaggatca cccatgtctg
      541 cagggccaag tggcaccgag tcggtgcttt ttttggatcc gcttggcgta actagatctt
      601 gagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag
      661 tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag aattacaaaa
      721 acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag atccactttg
      781 gcgccggctc gagggggccc gggtgcaaag atggataaag ttttaaacag agaggaatct
      841 ttgcagctaa tggaccttct aggtcttgaa aggagtggga attggctccg gtgcccgtca
      901 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
      961 atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     1021 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     1081 tctttttcgc aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg
     1141 gcctggcctc tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca
     1201 gtacgtgatt cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg
     1261 cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg
     1321 ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc
     1381 catttaaaat ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa
     1441 tgcgggccaa gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc
     1501 ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat
     1561 cggacggggg tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg
     1621 tatcgccccg ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag
     1681 atggccgctt cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga
     1741 gcgggcgggt gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc
     1801 atgtgactcc acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg
     1861 gagtacgtcg tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga
     1921 gtgggtggag actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc
     1981 cctttttgag tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt
     2041 tcttccattt caggtgtcgt gacgtacggc caccatgacc gagtacaagc ccacggtgcg
     2101 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     2161 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     2221 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     2281 cggcgccgcc gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     2341 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     2401 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     2461 agtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     2521 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     2581 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     2641 cacctggtgc atgacccgca agcccggtgc ctgaacgcgt taagtcgaca atcaacctct
     2701 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2761 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2821 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2881 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2941 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3001 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3061 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     3121 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3181 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3241 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
     3301 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     3361 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     3421 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     3481 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     3541 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca
     3601 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     3661 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3721 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3781 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     3841 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3901 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3961 ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     4021 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4081 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4141 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4201 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4261 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4321 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4381 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4441 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4501 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4561 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     4621 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4681 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4741 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4801 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4861 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4921 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4981 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5041 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5101 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5161 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5221 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5281 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5341 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5401 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5461 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5521 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5581 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5641 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5701 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5761 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5821 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5881 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5941 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6001 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6061 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6121 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6181 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6241 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6301 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6361 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6421 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6481 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6541 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6601 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6661 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6721 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6781 gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt
     6841 agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct
     6901 tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg
     6961 ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc
     7021 attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag
     7081 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     7141 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     7201 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     7261 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     7321 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     7381 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     7441 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     7501 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     7561 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     7621 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     7681 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     7741 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     7801 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     7861 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     7921 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
     7981 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     8041 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     8101 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     8161 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     8221 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
     8281 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
     8341 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaaca
//
