LOCUS       sequence-354503-        9298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral introduction of sgRNA constitute expression linked with 
            GFP marker into mammalian cell line..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9298)
  AUTHORS   Shi J, Wang E, Milazzo JP, Wang Z, Kinney JB, Vakoc CR
  TITLE     Discovery of cancer drug targets by CRISPR-Cas9 screening of protein
            domains.
  JOURNAL   Nat Biotechnol. 2015 May 11. doi: 10.1038/nbt.3235.
  PUBMED    25961408
REFERENCE   2  (bases 1 to 9298)
  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: "Nat 
            Biotechnol. 2015 May 11. doi: 10.1038/nbt.3235."
COMMENT     Sequence Label: LRG (Lenti_sgRNA_EFS_GFP)
FEATURES             Location/Qualifiers
     source          1..9298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     157..176
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     416..437
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(483..506)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      506..515
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(535..746)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    815..932
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(980..1055)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(2937..3185)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2995..3014)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(3165..3185)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(3363..3407)
                     /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    3592..3825
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    4318..4443
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             4490..4670
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(4671..4897)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(4925..4945)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4925..4943)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4964..4980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4964..4980)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4977..4999)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4988..5004
                     /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)."
     promoter        complement(join(5012..5018,5019..5036,5037..5042))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5057..5078
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5195..5212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5366..5954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5446..5465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6125..6916,6917..6985))
                     /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     6748..6767
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6986..7090)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7116..7571)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7253..7274)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7465..7484
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7698..7720
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7712..7729
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7713..7729
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7739..7758
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7739..7757
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      7778..7913
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    7953..8074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7964..7983)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     8018..8037
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             8152..8385
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    8457..9045
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             8563..8574
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     8972..8992
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(9061..9298,1..476,477..479,480..482))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(9064..9298,1..476,477..479,480..482))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9106..9127)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ccgttcttct gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc
       61 ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc
      121 accagggtgt cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg
      181 aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc
      241 tgcttcatgt ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg
      301 agggtgggcc agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg
      361 ccgtaggtgg catcgccctc gccctcgccg gacacgctga acttgtggcc gtttacgtcg
      421 ccgtccagct cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc
      481 atcttatcgt catcgtcttt gtaatccatg gtggcggatc ccgcgtcacg acacctgtgt
      541 tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac ttatatacgg
      601 ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt cccagtttac
      661 cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca acttctcggg
      721 gactgtgggc gatgtgcgct ctgcccactg acgggcaccg gagccactcg agccggcgcc
      781 aaagtggatc tctgctgtcc ctgtaataaa cccgaaaatt ttgaattttt gtaatttgtt
      841 tttgtaattc tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac
      901 tgtacccccc aatcccccct tttcttttaa aattgtggat gaatactgcc atttgtctca
      961 agatctagaa ttcaaaaaag caccgactcg gtgccacttt ttcaagttga taacggacta
     1021 gccttatttt aacttgctat ttctagctct aaaacagaga cgtacaaaaa agagcaagaa
     1081 gctaaaaaag atttaaaaat tatttttagc gcagttaatg gaacaggaac taaatttacc
     1141 ccaaaaatat tacgtgaatc aggatataac gttattgagg ttgaagagca tgcatttgaa
     1201 gatgaaacat ttaaaaatgt tgtaaatcca aatccagaat ttgatcctgc atgaaaaata
     1261 ccgcttgaat atggtattaa acatgatgca gatattatta ttatgaatga cccagatgct
     1321 gacagatttg gaatggcaat aaaacatgat ggtcattttg taagattaga tggaaatcaa
     1381 acaggaccaa ttttaattga ttgaaaatta tcaaatctaa aacgcttaaa tagcattcca
     1441 aaaaatccgg ctctatattc aagttttgta acaagtgatt tgggtgatag aatcgctcat
     1501 gaaaaatatg gagttaatat tgtaaaaact ttaactggat ttaaatgaat gggtagagaa
     1561 attgctaaag aagaagataa cggattaaat tttgtttttg cttatgaaga aagttatgga
     1621 tatgtaattg atgactcagc tagagataaa gatggaatac aagcttctat attaatagca
     1681 gaggctgctt gattttataa aaaacaaaat aaaacattag tagactattt agaagattta
     1741 tttaaagaaa tgggtgcata ttacactttc actttaaact tgaattttaa accagaagaa
     1801 aagaaattaa aaattgaacc attaatgaaa tcattgagag caacaccctt aactcaaatt
     1861 gctggactta aagttgttaa tgttgaagac tacatcgatg gaatgtataa tatgccagga
     1921 caagacttac taaaatttta tttagaagat aagtcatgat ttgctgttcg cccaagtgga
     1981 actgaaccta aactaaaaat ttattttata ggtgttggtg aatctgttca aaacgctaaa
     2041 gttaaagtag acgaaattat taaagaatta aaattaaaaa tgaatatata ggagaaaaaa
     2101 tgaaactaaa caaatatata gatcacacat tattaaaaca agatgctacg aaagctgaaa
     2161 ttaaacaatt atgtgatgaa gcaattgaat ttgattttgc aacagtttgt gttaattcat
     2221 attgaacaag ctattgtaaa gaattattaa aaggcacaaa tgtaggaata acaaatgttg
     2281 taggttttcc tctaggtgca tgcacaacag ctacaaaagc attcgaagtt tctgaagcaa
     2341 ttaaagatgg tgcaacagaa attgatatgg tattaaatat tggtgcatta aaagacaaaa
     2401 attatgaatt agttttagaa gacatgaaag ctgtaaaaaa agcagctgga tcacatgttg
     2461 ttaaatgtat tatggaaaat tgtttattaa caaaagaaga aatcatgaaa gcttgtgaaa
     2521 tagctgttga agctggatta gaatttgtta aaacatcaac aggattttca aaatcaggtg
     2581 caacatttga agatgttaaa ctaatgaagt cagttgttaa agacaatgct ttagttaaag
     2641 cagctggtgg agttagaaca tttgaagatg ctcaaaaaat gattgaagca ggagctgacc
     2701 gcttaggaac aagtggtgga gtagctatta ttaaaggtga agaaaacaac gcgagttact
     2761 aaaactagcg tttttttatt ttgctcattt ttattaaaag tttgcaaaaa ggaacataaa
     2821 aattctaatt attgatacta aagttattaa aaagaagatt ttggttgatt ttataaaggt
     2881 catagaatat aatattttag catgtgtatt ttgtgtgctc atttacaacc gtctccggtg
     2941 tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca agttacggta
     3001 agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc aagaaattat
     3061 tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa attaattcta
     3121 attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat gggaaatagg
     3181 ccctctctgg gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc
     3241 tgcctaactc tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct
     3301 cctactatca ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc
     3361 cacaaacttg cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt
     3421 tgcgattctt caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac
     3481 tccatccagg tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc
     3541 caaggcacag cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt
     3601 tgatccttta ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag
     3661 actgtgagtt gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc
     3721 tgcactatac cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc
     3781 atagtgcttc ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt
     3841 ttttctctct gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt
     3901 tttactactt tatatttata taattcactt ctccaattgt ccctcatatc tcctcctcca
     3961 ggtctgaaga tcagcggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc
     4021 tatcttgtct aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag
     4081 ggttgctact gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg
     4141 tagctgtccc agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc
     4201 gaatcgttct agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc
     4261 ccctggcctt aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac
     4321 gctctcgcac ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac
     4381 cagtcgccgc ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga
     4441 gagctcctct ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt
     4501 ttccacactg actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg
     4561 gcacacacta cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct
     4621 agttagccag agagctccca ggctcagatc tggtctaacc agagagaccc gtttatgtat
     4681 cgagctaggc acttaaatac aatatctctg caatgcggca attcagtggt tcgtccaatc
     4741 catgtcagac ccgtctgttg ccttcctaat aaggcacgat cgtaccacct tacttccacc
     4801 aatcggcatg cacggtgctt tttctctcct tgtaaggcat gttgctaact catcgttacc
     4861 atgttgcaag actacaagag tattgcataa gactacatta agcttgcagc tccagctttt
     4921 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     4981 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5041 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5101 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5161 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5221 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5281 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5341 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5401 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5461 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5521 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5581 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5641 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5701 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5761 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5821 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5881 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5941 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6001 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6061 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6121 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6181 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6241 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6301 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6361 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6421 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6481 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6541 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6601 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6661 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6721 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6781 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6841 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6901 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6961 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7021 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7081 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     7141 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     7201 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     7261 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     7321 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     7381 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     7441 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     7501 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     7561 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     7621 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     7681 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     7741 atacgactca ctatagggcg aattgggtac gtccctaggc ctccaaaaaa gcctcctcac
     7801 tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt
     7861 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gatgggcgga
     7921 gttaggggcg ggatagctag agccagacat gataagatac attgatgagt ttggacaaac
     7981 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     8041 atttgtaacc attataagct gcaataaaca agttcctctc actctctgat attcatttct
     8101 ttgcaagtta taaatactga ataataagat gacatgaact actatacgta ctgctagaga
     8161 ttttccacac tgactaaaag ggtctgaggg atctctagtt accagagtca cacaacagac
     8221 gggcacacac tacttgaagc actcaaggca agctttattg aggcttaagc agtgggttcc
     8281 ctagttagcc agagagctcc caggctcaga tctggtctaa ccagagagac ccagtacaag
     8341 caaaaagcag atcttgtctt cgttgggagt gaattagccc ttccagtccc cccttttctt
     8401 ttaaaaagtg gctaagatct acagctgcct tgtaagtcat tggtcttaaa gtcgacgcgg
     8461 ggaggcggcc caaagggaga tccgactcgt ctgagggcga aggcgaagac gcggaagagg
     8521 ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct ggattgaggg ccgaagggac
     8581 gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca caggcgagca gccaaggaaa
     8641 ggacgatgat ttccccgaca acaccacgga attgtcagtg cccaacagcc gagcccctgt
     8701 ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg gcaataggga gggggaaagc
     8761 gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc caaccagtgg gggttgcgtc
     8821 agcaaacaca gtgcacacca cgccacgttg cctgacaacg ggccacaact cctcataaag
     8881 agacagcaac caggatttat acaaggagga gaaaatgaaa gccatacggg aagcaatagc
     8941 atgatacaaa ggcattaaag cagcgtatcc acatagcgta aaaggagcaa catagttaag
     9001 aataccagtc aatctttcac aaattttgta atccagaggt tgattgtcga cttaacgcgt
     9061 ctacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     9121 gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta
     9181 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc
     9241 ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca ccttgatg
//
