LOCUS       sequence-210261-        9403 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral CRISPR-Cas9 Reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9403)
  AUTHORS   Labuhn M, Adams FF, Ng M, Knoess S, Schambach A, Charpentier EM, 
            Schwarzer A, Mateo JL, Klusmann JH, Heckl D
  TITLE     Refined sgRNA efficacy prediction improves large- and small-scale 
            CRISPR-Cas9 applications.
  JOURNAL   Nucleic Acids Res. 2018 Feb 16;46(3):1375-1385. doi: 
            10.1093/nar/gkx1268.
  PUBMED    29267886
REFERENCE   2  (bases 1 to 9403)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: "2018-02-16- 16"; volume: "46"; issue: 
            "3"; pages: "1375-1385"
COMMENT     Sequence Label: SIN40C.SFFV.sfGFP-RepEntry.iPAC
FEATURES             Location/Qualifiers
     source          1..9403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..118
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    235..256
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(271..276,277..294,295..301)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    309..325
                     /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     314..336
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     333..349
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     333..349
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     368..388
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        370..388
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        433..812
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        869..1060
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(892..912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1079..1305
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1306..1486
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1533..1658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2151..2384
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2569..2613
                     /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"
     misc_feature    2826..2943
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3001..3407
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3448..3457
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3520..4230
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             4261..4422
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     misc_feature    join(4449..5023,5024..5026,5027..5035)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4616..4633)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4843..4862
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5045..5644
                     /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(5045..5064)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5541..5561
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5671..6259
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5724..5744)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6142..6153)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6331..6564
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6642..6763
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6679..6698)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6733..6752
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6803..6938
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6958..6977)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6959..6977)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6987..7004)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6987..7003)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6996..7018)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7145..7600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7232..7251)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7442..7463
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7626..7730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7731..7799,7800..8591)
                     /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(7949..7968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8762..9350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9251..9270
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
       61 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      121 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      181 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      241 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      301 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      361 caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag cggtaccgtc
      421 gacctgcagg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
      481 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
      541 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
      601 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
      661 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
      721 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
      781 acatctacgt attagtcatc gctattacca tggcgcgcca tgcagggcct gaaataacct
      841 ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt
      901 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
      961 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     1021 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc ttaagaggcg cgccttaatg
     1081 tagtcttatg caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc
     1141 ttacaaggag agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt
     1201 gccttattag gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg
     1261 cattgcagag atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta
     1321 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     1381 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     1441 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     1501 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     1561 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     1621 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     1681 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     1741 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     1801 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     1861 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     1921 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     1981 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     2041 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     2101 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     2161 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     2221 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     2281 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     2341 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     2401 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     2461 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     2521 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     2581 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     2641 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     2701 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     2761 acccacctcc caaccccgag gggacaattc tagatcttga gacaaatggc agtattcatc
     2821 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     2881 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     2941 tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga gctagctgca
     3001 gtaacgccat tttgcaaggc atggaaaaat accaaaccaa gaatagagaa gttcagatca
     3061 agggcgggta catgaaaata gctaacgttg ggccaaacag gatatctgcg gtgagcagtt
     3121 tcggccccgg cccggggcca agaacagatg gtcaccgcag tttcggcccc ggcccgaggc
     3181 caagaacaga tggtccccag atatggccca accctcagca gtttcttaag acccatcaga
     3241 tgtttccagg ctcccccaag gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc
     3301 agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa aagagctcac
     3361 aacccctcac tcggcgcgcc agtcctccga cagactgagt cggccggtcg aatcaagctt
     3421 atcgataccg tcgacggatc caccggagcc accatgggaa ccggttatac gcgtggctct
     3481 ggatctggcg gcggaggaag cggaggcgga ggatctggat ctaagggcga ggaactgttc
     3541 accggcgtgg tgcccatcct ggtggaactg gatggcgacg tgaacggcca caagttctct
     3601 gtgcggggag agggcgaagg cgacgccaca aatggcaagc tgaccctgaa gttcatctgc
     3661 accaccggca agctgcccgt gccttggcct accctcgtga ccacactgac ctacggcgtg
     3721 cagtgcttca gcagataccc cgaccatatg aagcggcacg acttcttcaa gagcgccatg
     3781 cccgagggct acgtgcagga acggaccatc agcttcaagg acgacggcac ctacaagacc
     3841 agagccgaag tgaagttcga gggcgacacc ctcgtgaacc ggatcgagct gaagggcatc
     3901 gatttcaaag aggacggcaa catcctgggc cacaagctgg agtacaactt caacagccac
     3961 aatgtgtata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgg
     4021 cacaacgtgg aagatggcag cgtgcagctg gccgaccact accagcagaa cacccccatc
     4081 ggagatggcc ccgtgctgct gcccgacaac cactacctga gcacccagag cgtgctgagc
     4141 aaggacccca acgagaagcg ggaccacatg gtgctgctgg aattcgtgac cgccgctggc
     4201 atcacccacg gcatggacga gctgtataag ggctccggcg gaggctcccg gaccgaagag
     4261 tataagctga ttctgaacgg caagacactg aagggcgaga caaccaccga ggccgtggat
     4321 gccgccacag ccgagaaggt gttcaagcag tacgccaacg acaacggcgt ggacggcgag
     4381 tggacctacg acgatgccac caagaccttc accgtgaccg agtgaatgca tgagtaactg
     4441 aggatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     4501 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     4561 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     4621 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     4681 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     4741 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     4801 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     4861 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     4921 cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     4981 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccctggc
     5041 caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     5101 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     5161 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     5221 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     5281 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     5341 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5401 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5461 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5521 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     5581 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5641 ctgatgtaca agtgatcgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa
     5701 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5761 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5821 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5881 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5941 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     6001 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     6061 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     6121 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     6181 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     6241 ctttgggccg cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc
     6301 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     6361 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6421 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6481 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6541 agtcagtgtg gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt
     6601 atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt
     6661 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6721 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc
     6781 tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     6841 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     6901 gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc
     6961 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     7021 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     7081 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     7141 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     7201 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     7261 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     7321 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     7381 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     7441 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     7501 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7561 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     7621 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7681 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7741 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7801 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     7861 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     7921 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     7981 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     8041 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     8101 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     8161 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     8221 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     8281 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     8341 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     8401 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     8461 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     8521 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     8581 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     8641 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     8701 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     8761 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     8821 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     8881 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     8941 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     9001 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     9061 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     9121 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     9181 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     9241 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     9301 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     9361 acgcggcctt tttacggttc ctggcctttt gctggccttt tgc
//
