LOCUS       sequence-487495-        8298 bp    DNA     circular SYN 30-SEP-2025
DEFINITION  Backbone for the cloning of a dual sgRNA expression plasmid (hU6 and
            minimal H1/7SK hybrid promoters) using BsmbI. Selectable with a G418
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8298)
  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 8298)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 30, 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_H1/7SK
            hybrid)-EF1as_Thy...
FEATURES             Location/Qualifiers
     source          1..8298
                     /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"
     primer_bind     2006..2025
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_feature    2267..2384
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2453..2664
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2684..3169
                     /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             3179..3235
                     /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             3230..4030
                     /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(3290..3309)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3900..3919
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    4041..4629
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4094..4114)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4512..4523)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4701..4934
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5012..5133
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5049..5068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5103..5122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5173..5308
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5328..5347)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5329..5347)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5357..5374)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5357..5373)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5366..5388)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5515..5970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5602..5621)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5812..5833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5996..6100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6101..6169,6170..6961)
                     /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(6319..6338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7132..7720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7621..7640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7874..7891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8008..8029
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8044..8049,8050..8067,8068..8074)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8082..8098
                     /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     8087..8109
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8106..8122
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8106..8122
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8141..8161
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8143..8161
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(8189..8298,1..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 tccatattta gcatgtcgct atgtgttctg ggaaacttga cctaagtgta
     2041 aagttgagat ttccttcagg tttatatagt tctgtatgag accactcttt cccagagacg
     2101 ttcaattccc ggagagacac tcgaccgtct ctgtttaaga gctatgctgg aaacagcata
     2161 gcaagtttaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     2221 tttttgaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
     2281 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2341 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2401 acagcagaga tccactttgg cgccggctcg agtggctccg gtgcccgtca gtgggcagag
     2461 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
     2521 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     2581 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     2641 aacgggtttg ccgccagaac acaggtgtcg tgacgcggcc accatgaacc cagccatcag
     2701 cgtcgctctc ctgctctcag tcttgcaggt gtcccgaggg cagaaggtga ccagcctgac
     2761 agcctgcctg gtgaaccaaa accttcgcct ggactgccgc catgagaata acaccaagga
     2821 taactccatc cagcatgagt tcagcctgac ccgagagaag aggaagcacg tgctctcagg
     2881 caccctcggg atacccgagc acacgtaccg ctcccgcgtc accctctcca accagcccta
     2941 tatcaaggtc cttaccctag ccaacttcac caccaaggat gagggcgact acttttgtga
     3001 gcttcgagtc tcgggcgcga atcccatgag ctccaacaaa agtatcagtg tgtatagaga
     3061 caaactggtc aagtgtggcg gcataagcct gctggttcag aacacatcct ggatgctgct
     3121 gctgctgctt tccctctccc tcctccaagc cctggacttc atttctctgg gcagcggcgc
     3181 caccaacttc agcctgctga agcaggccgg cgacgtggag gagaaccccg gccccatgat
     3241 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3301 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3361 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3421 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3481 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3541 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3601 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3661 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3721 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3781 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3841 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3901 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3961 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4021 gttcttctga ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     4081 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     4141 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     4201 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     4261 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     4321 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     4381 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     4441 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     4501 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     4561 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     4621 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     4681 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     4741 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     4801 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     4861 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     4921 gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt atttataact
     4981 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     5041 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5101 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     5161 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5221 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5281 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     5341 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     5401 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5461 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     5521 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5581 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5641 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5701 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5761 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5821 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5881 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5941 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     6001 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6061 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6121 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6181 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6241 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6301 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6361 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6421 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6481 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6541 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6601 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6661 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     6721 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     6781 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     6841 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6901 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6961 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7021 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7081 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     7741 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     7801 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     7861 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     7921 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     7981 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     8041 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     8101 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     8161 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
     8221 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
     8281 cgattggtgg aagtaagg
//
