LOCUS       sequence-305801-        9431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-Cas9 Knock down.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9431)
  AUTHORS   Arnan C, Ullrich S, Pulido-Quetglas C, Nurtdinov R, Esteban A, 
            Blanco-Fernandez J, Aparicio-Prat E, Johnson R, Perez-Lluch S, Guigo
            R
  TITLE     Paired guide RNA CRISPR-Cas9 screening for protein-coding genes and 
            lncRNAs involved in transdifferentiation of human B-cells to 
            macrophages.
  JOURNAL   BMC Genomics. 2022 May 26;23(1):402. doi: 
            10.1186/s12864-022-08612-7.
  PUBMED    35619054
REFERENCE   2  (bases 1 to 9431)
  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: "BMC 
            Genomics."; date: "2022-05-26"; volume: "23(1):402. doi"; pages: " 
            10.1186/s12864-022-08612-7"
COMMENT     Sequence Label: pDECKO_mCherry_NFE2
FEATURES             Location/Qualifiers
     source          1..9431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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        1486..1734
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1486..1506
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1657..1676
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        1756..1831
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1838..2052
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     1973..1992
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     misc_RNA        2081..2156
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2212..2329
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2468..3646
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2699..3637
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3594..3614
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3659..4255
                     /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(3659..3678)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4155..4175
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4265..4318
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      4325..4334
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4331..5041
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4476..4494)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4744..4763
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    5057..5645
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5110..5130)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5528..5539)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5717..5950
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6028..6149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6065..6084)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6119..6138
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6189..6324
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6344..6363)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6345..6363)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6373..6390)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6373..6389)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6382..6404)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6531..6986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6618..6637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6828..6849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7012..7116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7117..7185,7186..7977)
                     /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(7335..7354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8148..8736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8637..8656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8890..8907
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9024..9045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9060..9065,9066..9083,9084..9090)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9098..9114
                     /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     9103..9125
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9122..9138
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9122..9138
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9157..9177
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9159..9177
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9205..9431
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccagagaggg cctatttccc
     1501 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tagaattaat
     1561 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     1621 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     1681 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccggaagt
     1741 gggaagccag aatctgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     1801 tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa cgctgacgtc atcaacccgc
     1861 tccaaggaat cgcgggccca gtgtcactag gcgggaacac ccagcgcgcg tgcgccctgg
     1921 caggaagatg gctgtgaggg acaggggagt ggcgccctgc aatatttgca tgtcgctatg
     1981 tgttctggga aatcaccata aacgtgaaat gtctttggat ttgggagtct tataagttct
     2041 gtatgagacc actctttccc ggaggtccaa ggtatggagc gttttagagc tagaaatagc
     2101 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     2161 ttaagcttgg cgtaactaga tcttgagaca aatggcagta ttcatccaca attttaaaag
     2221 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     2281 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     2341 cagggacagc agagatccac tttggcgccg gctcgagggg gcccgggtgc aaagatggat
     2401 aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt
     2461 gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     2521 gttgggggga ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg
     2581 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     2641 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt
     2701 aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc
     2761 ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag
     2821 tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga
     2881 ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct
     2941 cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt
     3001 tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt
     3061 tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg
     3121 cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct
     3181 ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc
     3241 ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa
     3301 atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc
     3361 ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca
     3421 cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta
     3481 tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt
     3541 gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc
     3601 tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta cggccaccat
     3661 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     3721 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     3781 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     3841 cggcaaggtg tgggtcgcgg acgacggcgc cgccgtggcg gtctggacca cgccggagag
     3901 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     3961 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     4021 cgcgtggttc ctggccaccg tcggagtctc gcccgaccac cagggcaagg gtctgggcag
     4081 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     4141 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     4201 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgccggatc
     4261 gggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccacc
     4321 ggtcgccacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
     4381 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     4441 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     4501 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     4561 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     4621 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     4681 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     4741 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     4801 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     4861 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     4921 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     4981 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta
     5041 aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5101 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5161 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     5221 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     5281 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     5341 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     5401 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     5461 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     5521 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     5581 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     5641 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     5701 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     5761 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     5821 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     5881 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     5941 atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca
     6001 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     6061 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6121 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     6181 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6241 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6301 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     6361 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     6421 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     6481 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     6541 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6601 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6661 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6721 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6781 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6841 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     6901 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6961 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     7021 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7081 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7141 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7201 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7261 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7321 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7381 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7441 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7501 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7561 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7621 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7681 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7741 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7801 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     7861 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     7921 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7981 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8041 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8101 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8161 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8221 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8281 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8341 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8401 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8461 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8521 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8581 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8641 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8701 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8761 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     8821 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     8881 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     8941 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     9001 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     9061 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     9121 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     9181 caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca
     9241 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
     9301 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
     9361 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
     9421 gatacataaa c
//
