pEF1a-PEmax-P2A-GFP

LOCUS       pEF1a-PEmax-P2A-GFP               11128 bp ds-DNA     circular SYN 08-JUN-2023
DEFINITION  Based on the pCMV-PEmax-P2A-GFP, we replaced the CMP promoter with 
            the EF1a promoter for expression in iPSC..
ACCESSION   .
VERSION     .
KEYWORDS    pEF1a-PEmax-P2A-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11128)
  TITLE     pEF1a-PEmax-P2A-GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11128)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 8, 2023 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..11128
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     3..22
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3..21
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             71..91
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             92..4192
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4250..4270
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6371..6391
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6401..6427
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6437..6493
                     /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             6494..7210
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6539..6560)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6800..6819)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7147..7168
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7219..7236
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7259..7276)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7265..7489
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7560..7576)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7560..7576)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7573..7595)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7584..7600
                     /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(7608..7638)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7653..7674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7791..7808)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7962..8550)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8042..8061)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8721..9581)
                     /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     9344..9363
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9582..9686)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9765..9784)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        9950..11128
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          10181..11119
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     11076..11096
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 gctaatacga ctcactatag ggagagccgc caccatgaaa cggacagccg acggaagcga
       61 gttcgagtca ccaaagaaga agcggaaagt cgacaagaag tacagcatcg gcctggacat
      121 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
      181 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
      241 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
      301 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
      361 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
      421 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
      481 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
      541 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
      601 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
      661 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat
      721 cctgtctgcc agactgagca agagcagaaa gctggaaaat ctgatcgccc agctgcccgg
      781 cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa
      841 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta
      901 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct
      961 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga
     1021 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga
     1081 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt
     1141 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga
     1201 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt
     1261 gaagctgaag agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc
     1321 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc
     1381 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta
     1441 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga
     1501 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt
     1561 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca
     1621 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac
     1681 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct
     1741 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa
     1801 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct
     1861 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga
     1921 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat
     1981 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct
     2041 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg
     2101 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag
     2161 aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc
     2221 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc
     2281 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat
     2341 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca
     2401 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct
     2461 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct
     2521 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa
     2581 ccggctgtcc gactacgatg tggacgctat cgtgcctcag agctttctga aggacgactc
     2641 catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc
     2701 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct
     2761 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact
     2821 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt
     2881 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg
     2941 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca
     3001 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc
     3061 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg
     3121 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa
     3181 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac
     3241 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga
     3301 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca
     3361 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct
     3421 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta
     3481 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa
     3541 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag
     3601 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa
     3661 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa
     3721 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa
     3781 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga
     3841 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga
     3901 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct
     3961 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca
     4021 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat
     4081 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag
     4141 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggtg actccggcgg
     4201 aagctctggt ggcagcaagc ggaccgccga cggctctgaa ttcgagagcc ctaagaagaa
     4261 aagaaaggtg agcggaggct ctagcggcgg aagcaccctg aacattgaag acgagtatag
     4321 actgcatgaa acaagcaagg aacccgacgt gtccctgggc tccacctggc tgtccgactt
     4381 tccccaggcc tgggccgaga caggaggaat gggcctggcc gtgcggcagg cacccctgat
     4441 catccctctg aaggccacct ctacacccgt gagcatcaag cagtacccta tgtctcagga
     4501 ggccagactg ggcatcaagc ctcacatcca gaggctgctg gaccagggca tcctggtgcc
     4561 atgccagagc ccctggaaca caccactgct gcccgtgaag aagccaggca ccaatgacta
     4621 tagacccgtg caggatctga gagaggtgaa caagagggtg gaggatatcc accccaccgt
     4681 gcccaaccct tacaatctgc tgtccggcct gcccccttct caccagtggt atacagtgct
     4741 ggacctgaag gatgccttct tttgtctgag actgcaccct accagccagc cactgttcgc
     4801 ctttgagtgg agggaccctg agatgggcat ctctggccag ctgacctgga cacgcctgcc
     4861 tcagggcttc aagaatagcc caacactgtt taacgaggcc ctgcaccgcg acctggcaga
     4921 tttccggatc cagcacccag atctgatcct gctgcagtac gtggacgatc tgctgctggc
     4981 cgccaccagc gagctggatt gccagcaggg aacacgcgcc ctgctgcaga ccctgggaaa
     5041 cctgggatat agggcatccg ccaagaaggc ccagatctgt cagaagcagg tgaagtacct
     5101 gggctatctg ctgaaggagg gccagagatg gctgacagag gccaggaagg agacagtgat
     5161 gggccagcca acacccaaga ccccaagaca gctgagggag ttcctgggca aagcaggatt
     5221 ttgcaggctg ttcatcccag gattcgcaga gatggcagca cctctgtacc cactgaccaa
     5281 gccgggcacc ctgtttaatt ggggccctga ccagcagaag gcctatcagg agatcaagca
     5341 ggccctgctg acagcaccag ccctgggcct gccagacctg accaagcctt tcgagctgtt
     5401 tgtggatgag aagcagggct acgccaaggg cgtgctgacc cagaagctgg gaccatggag
     5461 acggcccgtg gcctatctgt ccaagaagct ggacccagtg gcagcaggat ggccaccatg
     5521 cctgaggatg gtggcagcaa tcgccgtgct gacaaaggat gccggcaagc tgaccatggg
     5581 acagccactg gtcatcctgg caccacacgc agtggaggcc ctggtgaagc agcctccaga
     5641 tcgctggctg tctaacgccc ggatgacaca ctaccaggcc ctgctgctgg acaccgatcg
     5701 cgtgcagttt ggccctgtgg tggccctgaa tccagccacc ctgctgcctc tgccagagga
     5761 gggcctgcag cacaactgtc tggacatcct ggcagaggca cacggaacaa ggccagacct
     5821 gaccgatcag cccctgcctg acgccgatca cacatggtat accgatggaa gctccctgct
     5881 gcaggagggc cagaggaagg caggagcagc agtgaccaca gagacagaag tgatctgggc
     5941 caaggccctg ccagcaggca catccgccca gcgggccgag ctgatcgccc tgacccaggc
     6001 cctgaagatg gccgagggca agaagctgaa cgtgtacaca gactccagat atgccttcgc
     6061 caccgcacac atccacggag agatctacag gcgccggggc tggctgacct ctgagggcaa
     6121 ggagatcaag aacaaggatg agatcctggc cctgctgaag gccctgtttc tgcccaagcg
     6181 gctgagcatc atccactgtc ctggacacca gaagggacac tccgccgagg caaggggcaa
     6241 tcggatggcc gaccaggccg ccagaaaggc tgctattact gaaactcccg acacttccac
     6301 tctgctgatt gaaaactcct ccccttctgg cggctcaaaa agaaccgccg acggcagcga
     6361 attcgagtct cccaagaaga agaggaaagt cggctctggc cctgccgcta agagagtgaa
     6421 gctggacgga tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga
     6481 gaatcctgga ccgatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     6541 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     6601 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     6661 gccctggccc accctcgtga ccaccctgac ctatggagtg cagtgcttca gccgctaccc
     6721 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     6781 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     6841 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     6901 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     6961 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     7021 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     7081 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     7141 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     7201 gctgtacaag taaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
     7261 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     7321 accctggaag gtgccactcc cactgtcctt tcctaataaa atgagaaaat tgcatcgcat
     7381 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     7441 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     7501 gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg
     7561 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     7621 ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg
     7681 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     7741 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7801 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7861 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7921 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7981 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8041 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8101 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8161 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8221 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8281 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8341 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8401 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8461 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8521 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8581 gacactcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8641 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8701 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8761 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8821 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8881 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8941 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9001 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9061 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9121 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9181 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9241 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9301 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     9361 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     9421 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     9481 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     9541 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     9601 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     9661 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     9721 ggatcgggag atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat
     9781 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     9841 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     9901 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatag gctccggtgc
     9961 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
    10021 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
    10081 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
    10141 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
    10201 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
    10261 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
    10321 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
    10381 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
    10441 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
    10501 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
    10561 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
    10621 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
    10681 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
    10741 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
    10801 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
    10861 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
    10921 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
    10981 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
    11041 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
    11101 tttttttctt ccatttcagg tgtcgtga
//