LOCUS       sequence-226901-       10092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for Nkx2-5 and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10092)
  AUTHORS   Yu JSL, Palano G, Lim C, Moggio A, Drowley L, Plowright AT, 
            Bohlooly-Y M, Rosen BS, Hansson EM, Wang QD, Yusa K
  TITLE     CRISPR-KO screen identifies Dmap1 as a regulator of 
            chemically-induced reprogramming and differentiation of cardiac 
            progenitors.
  JOURNAL   Stem Cells. 2019 Apr 1. doi: 10.1002/stem.3012.
  PUBMED    30932271
REFERENCE   2  (bases 1 to 10092)
  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: "Stem Cells.
            2019 Apr 1. doi: 10.1002/stem.3012."
COMMENT     Sequence Label: pKLV2-U6(BsmBI)-EF1a-BFP-Nkx2-5
FEATURES             Location/Qualifiers
     source          1..10092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        960..1158
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1113..1133
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1176..1356
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1403..1528
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2027..2260
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2445..2489
                     /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    2787..2904
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2979..3227
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2979..2999
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3150..3169
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        3761..4942
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3992..4934
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     4891..4911
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      4955..4964
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4961..5662
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MGSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIK
                     VVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMAL
                     KLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARY
                     CDLPSKLGHKLN"
     CDS             5681..5734
                     /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"
     primer_bind     complement(6767..6787)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7185..7196)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7648..7665)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7654..7878
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7904..7924)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7904..7922)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7943..7959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7943..7959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7956..7978)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7967..7983
                     /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(7991..7997,7998..8015,8016..8021))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8036..8057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8174..8191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8345..8933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8425..8444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9104..9895,9896..9964))
                     /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     9727..9746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9965..10069)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgactagtac
      661 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      721 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      781 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      841 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      901 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      961 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1021 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1081 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1141 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     1201 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     1261 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     1321 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     1381 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     1441 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     1501 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1561 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1621 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1681 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1741 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1801 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1861 gcaagcggcc ggccgcgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1921 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1981 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     2041 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     2101 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     2161 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     2221 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     2281 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     2341 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2401 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2461 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2521 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2581 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2641 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2701 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2761 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2821 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2881 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2941 ttagtaccgg gccctacgcg tccaaggtcg ggcaggaaga gggcctattt cccatgattc
     3001 cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg
     3061 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     3121 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     3181 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgga gacgcatgac
     3241 tcgtctcagt ttaagagcta tgctggaaac agcatagcaa gtttaaataa ggctagtccg
     3301 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tacgcgtgag gggcgcggaa
     3361 gccggggcgg aggaggagcc aaccccgggc tgggctgaga cccgcagagg aagacgctct
     3421 agggatttgt cccggactag cgagatggca aggctgagga cgggaggctg attgagaggc
     3481 gaaggtacac cctaatctca atacaacctt tggagctaag ccagcaatgg tagagggaag
     3541 attctgcacg tcccttccag gcggcctccc cgtcaccacc ccccccaacc cgccccgacc
     3601 ggagctgaga gtaattcata caaaaggact cgcccctgcc ttggggaatc ccagggaccg
     3661 tcgttaaact cccactaacg tagaacccag agatcgctgc gttcccgccc cctcacccgc
     3721 ccgctctcgt catcactgag gtggagaaga gcatgcgtga ggctccggtg cccgtcagtg
     3781 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
     3841 cggtgcctag agaaagtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     3901 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     3961 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
     4021 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacgc ccctggctgc
     4081 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
     4141 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
     4201 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
     4261 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
     4321 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
     4381 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     4441 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
     4501 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     4561 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
     4621 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     4681 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     4741 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     4801 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     4861 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     4921 ttcttccatt tcaggtgtcg tggaagcttg gcgcgccacc atgggcagcg agctgattaa
     4981 ggagaacatg cacatgaagc tgtacatgga gggcaccgtg gacaaccatc acttcaagtg
     5041 cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa tcaaggtggt
     5101 cgagggcggc cctctcccct tcgccttcga catcctggct actagcttcc tctacggcag
     5161 caagaccttc atcaaccaca cccagggcat ccccgacttc ttcaagcagt ccttccctga
     5221 gggcttcaca tgggagagag tcaccacata cgaggacggg ggcgtgctga ccgctaccca
     5281 ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag gggtgaactt
     5341 cacatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct tcaccgagac
     5401 cctgtacccc gctgacggcg gcctggaagg cagaaacgac atggccctga agctcgtggg
     5461 cgggagccat ctgatcgcaa acatcaagac cacatataga tccaagaaac ccgctaagaa
     5521 cctcaagatg cctggcgtct actatgtgga ctacagactg gaaagaatca aggaggccaa
     5581 caacgagacc tacgtcgagc agcacgaggt ggcagtggcc agatactgcg acctccctag
     5641 caaactgggg cacaagctta atggaggggg cagcggcggg gaaggaaggg gctccctgct
     5701 cacctgcggc gacgtggaag agaaccccgg acctgtcgac gaattcatgt tccccagccc
     5761 tgcgctcaca cccacgcctt tctcagtcaa agacatcctg aacctggagc agcagcagcg
     5821 tagcctggcg tctggggacc tgtctgcgcg cctcgaggcc accctggccc ctgcctcctg
     5881 catgctggcc gccttcaagc ccgaggccta ctctggcccc gaggcggcag cgtccggcct
     5941 ggcagagctg cgcgcggaga tgggccccgc gccttcgccc cccaagtgct ctcctgcttt
     6001 cccagccgcc cccacatttt acccgggagc ctacggtgac cctgacccag ccaaagaccc
     6061 tcgggcggat aaaaaagagc tgtgcgcgct gcagaaggca gtggagctgg acaaagccga
     6121 gacggatggc gccgagagac cacgcgcacg gcggcgacgg aagccacgcg tgctcttctc
     6181 gcaggcgcag gtctacgagc tggagcggcg cttcaagcaa cagcggtacc tgtcggcgcc
     6241 agagcgcgac cagctggcca gcgtgctgaa gctcacgtcc acgcaggtca agatctggtt
     6301 ccagaaccgt cgctacaagt gcaagcgaca gcggcaggac cagactctgg agcttctggg
     6361 gccgccgccg ccgcccgcgc gcaggatcgc ggtgcccgtg ctggtgcgcg acgggaagcc
     6421 ctgcctgggg gaccccgcgg cctacgctcc cgcctacggc gtgggtctca atgcctatgg
     6481 ctacaacgcc tacccctacc ccagctacgg cggcgcggcc tgcagtcccg gctacagctg
     6541 cgccgcctac cccgctgcgc cccccgccgc gcagcccccc gccgcctccg ccaacagcaa
     6601 cttcgtgaac tttggcgtcg gggacttgaa caccgtgcag agtcccggga tgccgcaggg
     6661 caattcgggc gtctccacgc tgcacggcat ccgagcctgg taggctagcg gccgctcaac
     6721 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     6781 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     6841 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6901 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6961 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     7021 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     7081 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     7141 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     7201 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     7261 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcaggtacct ttaagaccaa
     7321 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     7381 ggctaattca ctcccaaaga agtcaagatc tgctttttgc ctgtactggg tctctctggt
     7441 tagaccagag tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     7501 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     7561 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     7621 ctctagcagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     7681 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7741 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7801 gtggggtggg gcaggacagc aagggggagg attgggaagt caatagcagg catgctgggg
     7861 atgcggtggg ctctatgggc ggccgttaat taagcttttg ttccctttag tgagggttaa
     7921 ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7981 caattccaca caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag
     8041 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     8101 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8161 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8221 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8281 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     8341 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     8581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8761 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     8821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8881 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8941 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     9001 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     9061 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     9121 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     9181 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     9241 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     9301 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     9361 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     9421 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     9481 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     9541 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     9601 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     9661 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     9721 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     9781 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     9841 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9901 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9961 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    10021 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    10081 gaaaagtgcc ac
//
