LOCUS       sequence-166334-        8435 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Backbone for CRISPR/Cas9 screening with single cell RNA-seq. 
            Lentiviral plasmid for cloning of gRNAs and Unique Guide Index 
            (UGI), with an mCherry fluorescent marker. Does NOT include Cas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8435)
  AUTHORS   Jaitin DA, Weiner A, Yofe I, Lara-Astiaso D, Keren-Shaul H, David E,
            Salame TM, Tanay A, van Oudenaarden A, Amit I
  TITLE     Dissecting Immune Circuits by Linking CRISPR-Pooled Screens with 
            Single-Cell RNA-Seq.
  JOURNAL   Cell. 2016 Dec 15;167(7):1883-1896.e15. doi: 
            10.1016/j.cell.2016.11.039.
  PUBMED    27984734
REFERENCE   2  (bases 1 to 8435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2016.11"; journalName: "Cell"; date: "2016-12-15- 
            15"; volume: "167"; issue: "7"; pages: "1883-1896.e15"
COMMENT     Sequence Label: CRISPseq-mCherry-backbone
FEATURES             Location/Qualifiers
     source          1..8435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..97
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    214..235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(250..255,256..273,274..280)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    288..304
                     /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     293..315
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     312..328
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     312..328
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     347..367
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        349..367
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        395..621
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             622..802
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    849..974
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1467..1700
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1885..1929
                     /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        2107..2355
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2107..2127
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2278..2297
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2372..2447
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2503..2620
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2759..3936
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2990..3927
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3884..3904
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3943..3952
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3949..4659
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4094..4112)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4362..4381
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4680..5268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4733..4753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5151..5162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5342..5575
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5653..5774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5690..5709)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5744..5763
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5814..5949
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5969..5988)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5970..5988)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5998..6015)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5998..6014)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6007..6029)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6156..6611
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6243..6262)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6453..6474
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6637..6741
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6742..6810,6811..7602)
                     /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(6960..6979)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7773..8361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8262..8281
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
       61 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
      121 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      181 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
      241 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      301 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
      361 ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag
      421 tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt
      481 gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg
      541 ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag
      601 tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc
      661 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
      721 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
      781 tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac
      841 cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg
      901 ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg
      961 ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt
     1021 taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc
     1081 tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac
     1141 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     1201 cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt
     1261 tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg
     1321 atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat
     1381 aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg
     1441 cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca
     1501 ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct
     1561 ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg
     1621 caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac
     1681 ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact
     1741 gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg
     1801 acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt
     1861 gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg
     1921 gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa attattcata
     1981 atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat
     2041 agagttaggc agggatattc accattatcg tttcagaccc acctcccaac cccgagggga
     2101 cccagagagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     2161 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     2221 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     2281 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     2341 aaaggacgaa acaccggaga cggacgtctc tgttttagag ctagaaatag caagttaaaa
     2401 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttaagcttg
     2461 gcgtaactag atcttgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2521 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2581 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2641 cagagatcca ctttggcgcc ggctcgaggg ggcccgggtg caaagatgga taaagtttta
     2701 aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaattgg
     2761 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     2821 aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     2881 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     2941 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt
     3001 gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac
     3061 ttccactggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
     3121 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     3181 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     3241 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     3301 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     3361 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     3421 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
     3481 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     3541 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     3601 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     3661 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     3721 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     3781 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     3841 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     3901 gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta cggccaccat ggtgagcaag
     3961 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     4021 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     4081 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     4141 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     4201 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     4261 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     4321 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     4381 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     4441 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     4501 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     4561 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     4621 cgccactcca ccggcggcat ggacgagctg tacaagtaaa cgcgttaagt cgagtcgaca
     4681 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4741 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4801 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4861 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4921 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4981 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5041 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     5101 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5161 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5221 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgacggatcc
     5281 ggtgatggga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     5341 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     5401 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5461 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5521 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     5581 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     5641 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5701 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5761 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     5821 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5881 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     5941 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     6001 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     6061 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     6121 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     6181 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     6241 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6301 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6361 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6421 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6481 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6541 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6601 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6661 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6721 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6781 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6841 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6901 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6961 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     7021 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     7081 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     7141 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7201 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7261 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7321 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7381 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7441 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7501 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7561 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7621 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7681 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7741 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7801 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7861 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7921 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7981 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     8041 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     8101 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     8161 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     8221 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8281 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8341 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8401 tgctggcctt ttgctcacat gttctttcct gcgtt
//
