LOCUS       sequence-456127-       10746 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Small lentiviral backbone for CRISPR guide libraries, improved titre
            for hard to transduce cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10746)
  TITLE     pLentiCRISPR_mCherry
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10746)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pLentiCRISPR_mCherry
FEATURES             Location/Qualifiers
     source          1..10746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2371..2619
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2371..2391
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2542..2561
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4501..4576
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4638..4849
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4863..4872
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4878..5585
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(5020..5038)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5288..5307
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5601..6189
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5654..5674)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6072..6083)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6261..6494
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6520..6537)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6526..6750
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6796..7224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6883..6902)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7093..7114
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7233..7253)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7238..7567
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7418..7553
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7480..7499
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7615..7662
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7681..8055
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    8185..8306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8222..8241)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8276..8295
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8355..8371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8355..8371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8368..8390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8379..8395
                     /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(8403..8409,8410..8427,8428..8433))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8448..8469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8586..8603)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8757..9345)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8837..8856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9516..10307,10308..10376))
                     /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     10139..10158
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10377..10481)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10556..10575)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaaggtacc gagggcctat ttcccatgat tccttcatat
     2401 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     2461 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     2521 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
     2581 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gagacggttg taaatgagca
     2641 cacaaaatac acatgctaaa atattatatt ctatgacctt tataaaatca accaaaatct
     2701 tctttttaat aactttagta tcaataatta gaatttttat gttccttttt gcaaactttt
     2761 aataaaaatg agcaaaataa aaaaacgcta gttttagtaa ctcgcgttgt tttcttcacc
     2821 tttaataata gctactccac cacttgttcc taagcggtca gctcctgctt caatcatttt
     2881 ttgagcatct tcaaatgttc taactccacc agctgcttta actaaagcat tgtctttaac
     2941 aactgacttc attagtttaa catcttcaaa tgttgcacct gattttgaaa atcctgttga
     3001 tgttttaaca aattctaatc cagcttcaac agctatttca caagctttca tgatttcttc
     3061 ttttgttaat aaacaatttt ccataataca tttaacaaca tgtgatccag ctgctttttt
     3121 tacagctttc atgtcttcta aaactaattc ataatttttg tcttttaatg caccaatatt
     3181 taataccata tcaatttctg ttgcaccatc tttaattgct tcagaaactt cgaatgcttt
     3241 tgtagctgtt gtgcatgcac ctagaggaaa acctacaaca tttgttattc ctacatttgt
     3301 gccttttaat aattctttac aatagcttgt tcaatatgaa ttaacacaaa ctgttgcaaa
     3361 atcaaattca attgcttcat cacataattg tttaatttca gctttcgtag catcttgttt
     3421 taataatgtg tgatctatat atttgtttag tttcattttt tctcctatat attcattttt
     3481 aattttaatt ctttaataat ttcgtctact ttaactttag cgttttgaac agattcacca
     3541 acacctataa aataaatttt tagtttaggt tcagttccac ttgggcgaac agcaaatcat
     3601 gacttatctt ctaaataaaa ttttagtaag tcttgtcctg gcatattata cattccatcg
     3661 atgtagtctt caacattaac aactttaagt ccagcaattt gagttaaggg tgttgctctc
     3721 aatgatttca ttaatggttc aatttttaat ttcttttctt ctggtttaaa attcaagttt
     3781 aaagtgaaag tgtaatatgc acccatttct ttaaataaat cttctaaata gtctactaat
     3841 gttttatttt gttttttata aaatcaagca gcctctgcta ttaatataga agcttgtatt
     3901 ccatctttat ctctagctga gtcatcaatt acatatccat aactttcttc ataagcaaaa
     3961 acaaaattta atccgttatc ttcttcttta gcaatttctc tacccattca tttaaatcca
     4021 gttaaagttt ttacaatatt aactccatat ttttcatgag cgattctatc acccaaatca
     4081 cttgttacaa aacttgaata tagagccgga ttttttggaa tgctatttaa gcgttttaga
     4141 tttgataatt ttcaatcaat taaaattggt cctgtttgat ttccatctaa tcttacaaaa
     4201 tgaccatcat gttttattgc cattccaaat ctgtcagcat ctgggtcatt cataataata
     4261 atatctgcat catgtttaat accatattca agcggtattt ttcatgcagg atcaaattct
     4321 ggatttggat ttacaacatt tttaaatgtt tcatcttcaa atgcatgctc ttcaacctca
     4381 ataacgttat atcctgattc acgtaatatt tttggggtaa atttagttcc tgttccatta
     4441 actgcgctaa aaataatttt taaatctttt ttagcttctt gctctttttt gtacgtctct
     4501 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
     4561 ggcaccgagt cggtgctttt ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg
     4621 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     4681 aggggtcggc aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     4741 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     4801 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg accggtctag
     4861 atgccaccat ggttccggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     4921 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     4981 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     5041 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     5101 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     5161 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     5221 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     5281 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     5341 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     5401 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     5461 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     5521 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     5581 agtaaacgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     5641 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     5701 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     5761 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     5821 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     5881 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     5941 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     6001 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     6061 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6121 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     6181 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     6241 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     6301 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6361 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6421 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6481 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     6541 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     6601 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     6661 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     6721 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     6781 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6841 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6901 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6961 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     7021 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7081 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     7141 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     7201 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     7261 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     7321 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     7381 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     7441 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     7501 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     7561 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     7621 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     7681 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     7741 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     7801 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     7861 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     7921 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     7981 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     8041 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     8101 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     8161 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     8221 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     8281 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     8341 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     8401 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     8461 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     8521 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     8581 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     8641 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     8701 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     8761 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     8821 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8881 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8941 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     9001 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     9061 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     9121 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     9181 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     9241 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     9301 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     9361 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     9421 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     9481 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     9541 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     9601 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     9661 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     9721 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     9781 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     9841 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     9901 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     9961 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
    10021 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
    10081 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    10141 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    10201 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    10261 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    10321 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
    10381 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
    10441 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
    10501 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
    10561 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
    10621 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
    10681 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
    10741 cgcgtt
//
