LOCUS       sequence-486276-        8421 bp    DNA     circular SYN 16-SEP-2025
DEFINITION  Backbone for the cloning of a dual sgRNA expression plasmid (hU6 and
            H1 promoters) using BsmbI. Selectable with a G418 resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8421)
  AUTHORS   Stadager J, Bernardini C, Hartmann L, May H, Wiepcke J, Kuban M, 
            Najafova Z, Johnsen SA, Legewie S, Traube FR, Jude J, Rathert P
  TITLE     CRISPR GENome and epigenome engineering improves loss-of-function 
            genetic-screening approaches.
  JOURNAL   Cell Rep Methods. 2025 Jun 16;5(6):101078. doi: 
            10.1016/j.crmeth.2025.101078. Epub 2025 Jun 10.
  PUBMED    40499551
REFERENCE   2  (bases 1 to 8421)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep 
            Methods."; date: "2025-06-16"; volume: "5(6):101078. doi"; pages: " 
            10.1016/j.crmeth.2025.101078. Epub 2025 Jun 10"
COMMENT     Sequence Label: sgRNA-Dual_filler(hU6_H1)-EF1a-Thy1.1-P2A-Neo
FEATURES             Location/Qualifiers
     source          1..8421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             118..298
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    345..470
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    963..1196
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1381..1425
                     /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        1603..1851
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1603..1623
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1774..1793
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        1994..2208
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     2129..2148
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_feature    2390..2507
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2576..2787
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2807..3292
                     /codon_start=1
                     /product="mouse GPI-anchored cell surface glycoprotein of 
                     the immunoglobulin superfamily"
                     /label=Thy1.1
                     /note="This allele of Thy1 is also known as CD90.1."
                     /translation="MNPAISVALLLSVLQVSRGQKVTSLTACLVNQNLRLDCRHENNTK
                     DNSIQHEFSLTREKRKHVLSGTLGIPEHTYRSRVTLSNQPYIKVLTLANFTTKDEGDYF
                     CELRVSGANPMSSNKSISVYRDKLVKCGGISLLVQNTSWMLLLLLSLSLLQALDFISL"
     CDS             3302..3358
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3353..4153
                     /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="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3413..3432)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4023..4042
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    4164..4752
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4217..4237)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4635..4646)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4824..5057
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5135..5256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5172..5191)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5226..5245
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5296..5431
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5451..5470)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5452..5470)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5480..5497)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5480..5496)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5489..5511)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5638..6093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5725..5744)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5935..5956
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6119..6223
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6224..6292,6293..7084)
                     /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(6442..6461)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7255..7843
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7744..7763
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7997..8014
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8131..8152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8167..8172,8173..8190,8191..8197)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8205..8221
                     /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     8210..8232
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8229..8245
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8229..8245
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8264..8284
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8266..8284
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(8312..8421,1..117)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc
       61 actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg
      121 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      181 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      241 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      301 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      361 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      421 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      481 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
      541 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
      601 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
      661 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
      721 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
      781 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
      841 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
      901 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
      961 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1021 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1081 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1141 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1201 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1261 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1321 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1381 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1441 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1501 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     1561 ttatcgtttc agacccacct cccaaccccg aggggaccca gagagggcct atttcccatg
     1621 attccttcat atttgcatat acgatacaag gctgttagag agataattag aattaatttg
     1681 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
     1741 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
     1801 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cagagacggt
     1861 tgtaaatgag cacacaaaat acacatgcgt ctccgagagc taagcagaaa gctgcatagc
     1921 aagttcaaat aaggctagtc cgtacacaac ttgaaaaagt ggcagccgag tcggctgctt
     1981 ttttttcagc tccgaacgct gacgtcatca acccgctcca aggaatcgcg ggcccagtgt
     2041 cactaggcgg gaacacccag cgcgcgtgcg ccctggcagg aagatggctg tgagggacag
     2101 gggagtggcg ccctgcaata tttgcatgtc gctatgtgtt ctgggaaatc accataaacg
     2161 tgaaatgtct ttggatttgg gaatcttata agttctgtat gagaccactc tttcccagag
     2221 acgttcaatt cccggagaga cactcgaccg tctccgttta agagctatgc tggaaacagc
     2281 atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     2341 ctttttttga attctagatc ttgagacaaa tggcagtatt catccacaat tttaaaagaa
     2401 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     2461 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     2521 gggacagcag agatccactt tggcgccggc tcgagtggct ccggtgcccg tcagtgggca
     2581 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
     2641 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
     2701 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
     2761 cgcaacgggt ttgccgccag aacacaggtg tcgtgacgcg gccaccatga acccagccat
     2821 cagcgtcgct ctcctgctct cagtcttgca ggtgtcccga gggcagaagg tgaccagcct
     2881 gacagcctgc ctggtgaacc aaaaccttcg cctggactgc cgccatgaga ataacaccaa
     2941 ggataactcc atccagcatg agttcagcct gacccgagag aagaggaagc acgtgctctc
     3001 aggcaccctc gggatacccg agcacacgta ccgctcccgc gtcaccctct ccaaccagcc
     3061 ctatatcaag gtccttaccc tagccaactt caccaccaag gatgagggcg actacttttg
     3121 tgagcttcga gtctcgggcg cgaatcccat gagctccaac aaaagtatca gtgtgtatag
     3181 agacaaactg gtcaagtgtg gcggcataag cctgctggtt cagaacacat cctggatgct
     3241 gctgctgctg ctttccctct ccctcctcca agccctggac ttcatttctc tgggcagcgg
     3301 cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg gaggagaacc ccggccccat
     3361 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3421 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3481 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3541 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3601 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3661 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3721 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3781 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3841 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     3901 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3961 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4021 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4081 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4141 cgagttcttc tgattaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     4201 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4261 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4321 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4381 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4441 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4501 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     4561 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     4621 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4681 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4741 ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca
     4801 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     4861 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     4921 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     4981 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     5041 gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata
     5101 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     5161 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5221 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     5281 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5341 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     5401 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     5461 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     5521 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     5581 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     5641 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     5701 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     5761 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     5821 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     5881 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     5941 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     6001 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     6061 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     6121 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6181 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6241 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6301 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6361 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6421 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6481 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6541 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6601 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6661 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6721 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6781 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6841 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6901 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6961 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     7021 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7081 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     7141 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     7201 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     7261 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7321 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7381 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7441 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7501 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7561 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7621 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7681 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7741 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7801 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7861 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7921 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7981 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     8041 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     8101 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     8161 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     8221 atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta
     8281 aagggaacaa aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct
     8341 tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca
     8401 tgccgattgg tggaagtaag g
//
