LOCUS       sequence-217983-        8972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral U6 driven sgRNA cloning vector where guide sequences are 
            inserted between BfuAI sites, improved cassette cloning efficiency.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8972)
  TITLE     Improved U6-sgRNA vector for expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8972)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pLKO.1-puro U6 sgRNA BfuAI large stuffer
FEATURES             Location/Qualifiers
     source          1..8972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..111
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    228..249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     mobile_element  274..1041
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     promoter        join(1042..1046,1047..1064,1065..1071)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1079..1095
                     /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     1084..1106
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1103..1119
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1103..1119
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1138..1158
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1140..1158
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1186..1412
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1413..1593
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1640..1765
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2258..2491
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2676..2720
                     /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        3018..3266
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3018..3038
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3189..3208
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             complement(3397..4191)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3508..3527)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4118..4137
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     misc_RNA        4350..4425
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    4474..4591
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4640..5150
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     5069..5087
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             5172..5771
                     /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(5172..5191)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5668..5688
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5899..6132
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6204..6325
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6241..6260)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6295..6314
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6365..6500
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6520..6539)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6521..6539)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6549..6566)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6549..6565)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6558..6580)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6707..7162
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6794..6813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7004..7025
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7188..7292
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7293..7361,7362..8153)
                     /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(7511..7530)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8324..8912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8813..8832
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttggtaatg actccaactt attgatagtg
      301 ttttatgttc agataatgcc cgatgacttt gtcatgcagc tccaccgatt ttgagaacga
      361 cagcgacttc cgtcccagcc gtgccaggtg ctgcctcaga ttcaggttat gccgctcaat
      421 tcgctgcgta tatcgcttgc tgattacgtg cagctttccc ttcaggcggg attcatacag
      481 cggccagcca tccgtcatcc atatcaccac gtcaaagggt gacagcaggc tcataagacg
      541 ccccagcgtc gccatagtgc gttcaccgaa tacgtgcgca acaaccgtct tccggagact
      601 gtcatacgcg taaaacagcc agcgctggcg cgatttagcc ccgacatagc cccactgttc
      661 gtccatttcc gcgcagacga tgacgtcact gcccggctgt atgcgcgagg ttaccgactg
      721 cggcctgagt tttttaagtg acgtaaaatc gtgttgaggc caacgcccat aatgcgggct
      781 gttgcccggc atccaacgcc attcatggcc atatcaatga ttttctggtg cgtaccgggt
      841 tgagaagcgg tgtaagtgaa ctgcagttgc catgttttac ggcagtgaga gcagagatag
      901 cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc cgtcagtagc tgaacaggag
      961 ggacagctga tagaaacaga agccactgga gcacctcaaa aacaccatca tacactaaat
     1021 cagtaagttg gcagcatcac cttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     1081 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     1141 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     1201 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     1261 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     1321 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     1381 ttgtatttaa gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag
     1441 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     1501 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     1561 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     1621 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     1681 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     1741 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     1801 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     1861 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     1921 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1981 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     2041 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     2101 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     2161 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
     2221 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
     2281 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
     2341 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
     2401 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
     2461 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
     2521 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
     2581 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
     2641 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
     2701 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
     2761 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt
     2821 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa
     2881 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag
     2941 acagatccat tcgattagtg aacggatctc gacggtatcg atcacgagac tagcctcgag
     3001 cggccgcccc cttcaccgag ggcctatttc ccatgattcc ttcatatttg catatacgat
     3061 acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag atattagtac
     3121 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     3181 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     3241 atatcttgtg gaaaggacga aacaccggtg tgcaggtgag tgatccaaac gcccggcggc
     3301 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     3361 ggagtccaag cgagctctcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     3421 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     3481 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     3541 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     3601 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     3661 gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     3721 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     3781 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     3841 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     3901 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     3961 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt
     4021 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     4081 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     4141 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     4201 tcctcatcct gtctcttgat cagatcttga tcccctgcgc catcagatcc ttggcggcaa
     4261 gaaagccatc cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg
     4321 caattccgac ggatcaagaa cctgctgacg ttttagagct agaaatagca agttaaaata
     4381 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttgaattctc
     4441 gacctcgaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     4501 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     4561 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     4621 actttggccg cggctcgagg gggttggggt tgcgcctttt ccaaggcagc cctgggtttg
     4681 cgcagggacg cggctgctct gggcgtggtt ccgggaaacg cagcggcgcc gaccctgggt
     4741 ctcgcacatt cttcacgtcc gttcgcagcg tcacccggat cttcgccgct acccttgtgg
     4801 gccccccggc gacgcttcct gctccgcccc taagtcggga aggttccttg cggttcgcgg
     4861 cgtgccggac gtgacaaacg gaagccgcac gtctcactag taccctcgca gacggacagc
     4921 gccagggagc aatggcagcg cgccgaccgc gatgggctgt ggccaatagc ggctgctcag
     4981 cagggcgcgc cgagagcagc ggccgggaag gggcggtgcg ggaggcgggg tgtggggcgg
     5041 tagtgtgggc cctgttcctg cccgcgcggt gttccgcatt ctgcaagcct ccggagcgca
     5101 cgtcggcagt cggctccctc gttgaccgaa tcaccgacct ctctccccag ggggatccac
     5161 cggagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     5221 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     5281 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     5341 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     5401 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     5461 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     5521 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     5581 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     5641 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     5701 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     5761 ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc
     5821 atgcatcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     5881 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     5941 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6001 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6061 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6121 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6181 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     6241 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6301 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     6361 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6421 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6481 aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac
     6541 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     6601 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     6661 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     6721 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6781 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6841 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6901 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6961 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     7021 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     7081 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7141 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     7201 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     7261 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     7321 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     7381 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     7441 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     7501 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     7561 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7621 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7681 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7741 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7801 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7861 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7921 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7981 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     8041 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     8101 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     8161 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     8221 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     8281 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     8341 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8401 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8461 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8521 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8581 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8641 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8701 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8761 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8821 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     8881 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     8941 tcctggcctt ttgctggcct tttgctcaca tg
//
