LOCUS       sequence-414904-        9106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of single SpCas9 and AsCas12a gRNAs for 
            generating combinatorial CHyMErA 3Cs libraries.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9106)
  AUTHORS   Cetin R, Wegner M, Luwisch L, Saud S, Achmedov T, Susser S, 
            Vera-Guapi A, Muller K, Matthess Y, Quandt E, Schaubeck S, Beisel 
            CL, Kaulich M
  TITLE     Optimized metrics for orthogonal combinatorial CRISPR screens.
  JOURNAL   Sci Rep. 2023 May 6;13(1):7405. doi: 10.1038/s41598-023-34597-8.
  PUBMED    37149686
REFERENCE   2  (bases 1 to 9106)
  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: "Sci Rep."; 
            date: "2023-05-6"; volume: "13(1):7405. doi"; pages: " 
            10.1038/s41598-023-34597-8"
COMMENT     Sequence Label: 
            pLenti_CHyMErA_U6(SpCas9_I-SceI)-(AsCas12_PacI)_PGK-puro
FEATURES             Location/Qualifiers
     source          1..9106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1679..1912
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2097..2141
                     /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    2439..2556
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2614..2862
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2614..2634
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2785..2804
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2885..2960
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3044..3543
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3079..3102)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3476..3495
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3583..4182
                     /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(3583..3602)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4079..4099
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    4198..4786
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4251..4271)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4669..4680)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4858..5091
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5117..5134)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5123..5347
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5393..5821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5480..5499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5690..5711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5830..5850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5835..6164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6015..6150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6077..6096
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6212..6259
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6278..6652
                     /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    6782..6903
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6819..6838)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6873..6892
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6952..6968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6952..6968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6965..6987)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6976..6992
                     /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(7000..7006,7007..7024,7025..7030))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7045..7066
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7183..7200)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7354..7942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7434..7453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8113..8904,8905..8973))
                     /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     8736..8755
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8974..9078)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
     1561 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     1621 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     1681 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     1741 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     1801 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     1861 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     1921 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     1981 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     2041 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     2101 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     2161 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     2221 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     2281 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     2341 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg
     2401 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     2461 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2521 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2581 gatccagttt ggttagtacc gggccctggt accgagggcc tatttcccat gattccttca
     2641 tatttgcata tacgatacaa ggctgttaga gagataatta gaattaattt gactgtaaac
     2701 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
     2761 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
     2821 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggtatagg gataacaggg
     2881 taatgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
     2941 aagtggcacc gagtcggtgc taatttctac tcttgtagat tcgaattaat taatgttaat
     3001 taattttttg ctagcaagct tggcgtaagg atccaattct accgggtagg ggaggcgctt
     3061 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca
     3121 agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc
     3181 tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc
     3241 ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg
     3301 cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag
     3361 cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg
     3421 ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc
     3481 tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt
     3541 tcgacctgca tccatctaga tctcgagcag ctgaagctta ccatgaccga gtacaagccc
     3601 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3661 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3721 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3781 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3841 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3901 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3961 accgtcggag tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     4021 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     4081 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4141 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaacgcgtta agtcgacaat
     4201 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4261 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4321 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4381 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4441 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4501 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4561 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     4621 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4681 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4741 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcgtcg actttaagac
     4801 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     4861 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct
     4921 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     4981 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     5041 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agggccagtt
     5101 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     5161 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     5221 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     5281 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     5341 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     5401 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5461 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5521 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5581 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     5641 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5701 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     5761 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5821 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5881 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5941 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     6001 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6061 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     6121 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     6181 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     6241 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt
     6301 tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg
     6361 gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct
     6421 gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt
     6481 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     6541 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     6601 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct
     6661 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     6721 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     6781 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     6841 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     6901 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     6961 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     7021 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     7081 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7141 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7201 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7261 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7321 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7381 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7441 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7501 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7561 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7621 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7681 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7741 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7801 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7861 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7921 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7981 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     8041 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     8101 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8161 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8221 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8281 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8341 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8401 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8461 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8521 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8581 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8641 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8701 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8761 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8821 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8881 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8941 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     9001 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     9061 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac
//
