LOCUS       sequence-389659-        9800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of sfGFP WT with Methanosarcina barkeri (Mb) Pyl-tRNA for
            the encoding of non-canonical amino acids at TAG codons in HEK293 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9800)
  AUTHORS   Avila-Crump S, Hemshorn ML, Jones CM, Mbengi L, Meyer K, Griffis JA,
            Jana S, Petrina GE, Pagar VV, Karplus PA, Petersson EJ, Perona JJ, 
            Mehl RA, Cooley RB
  TITLE     Generating Efficient Methanomethylophilus alvus Pyrrolysyl-tRNA 
            Synthetases for Structurally Diverse Non-Canonical Amino Acids.
  JOURNAL   ACS Chem Biol. 2022 Dec 16;17(12):3458-3469. doi: 
            10.1021/acschembio.2c00639. Epub 2022 Nov 16.
  PUBMED    36383641
REFERENCE   2  (bases 1 to 9800)
  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"; doi: 
            "10.1021/acschembio.2c00639"; journalName: "ACS Chem Biol"; date: 
            "2022-12-16- 16"; volume: "17"; issue: "12"; pages: "3458-3469"
COMMENT     Sequence Label: pAcBac1-Mb-sfGFP WT
FEATURES             Location/Qualifiers
     source          1..9800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  83..248
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        complement(341..435)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    341..359
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    369..387
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(512..760)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     polyA_signal    793..927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(817..836)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     871..890
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1073..2560,2561..2608))
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein"
                     /label=VSV-G
                     /note="Indiana strain"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHNDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAVIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNYICPTVHNSTTWHSDYKVKGLCDSNLISMDITFFSEDGELSSLGKEGTGFR
                     SNYFAYETGGKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPAFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDESLFFGDTGLSKNPIELVEGWF
                     SSWKSSIASFFFIIGLIIGLFLVLRVGIHLCIKLKHTKKRQIYTDIEMNRLGK"
     promoter        complement(2667..2758)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(2725..2742)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     enhancer        2853..3232
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3233..3436
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3387..3407
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3481..3500
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3481..3499
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3522..3531
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3528..3530,3531..3533,3534..4244)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     CDS             4251..4292
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4293..4310
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    4336..4924
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4389..4409)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4807..4818)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4973..4990)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4979..5203
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        5240..5488
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        5565..5659
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    5613..5631
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5641..5659
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        join(6149..6154,6155..6171,6172..6177)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             6366..6899
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  6966..7190
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(7324..7341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7495..8083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7575..7594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8254..9045,9046..9114))
                     /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     8877..8896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9115..9219)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9245..9699)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9382..9403)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9593..9612
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aaagtagccg aagatgacgg tttgtcacat ggagttggca ggatgtttga ttaaaaacat
       61 aacaggaaga aaaatgcccc gctgtgggcg gacaaaatag ttgggaactg ggaggggtgg
      121 aaatggagtt tttaaggatt atttagggaa gagtgacaaa atagatggga actgggtgta
      181 gcgtcgtaag ctaatacgaa aattaaaaat gacaaaatag tttggaacta gatttcactt
      241 atctggttcg gatctcctag caaaaaacgg ggggtgcggg agtttcaccc gcctgcacgg
      301 atttagagtc cgtgtgatct attcgatcca ccccccggtg tctctatcac tgatagggaa
      361 cttataagtc tctatcactg atagggattt cacgtttatg gtgatttccc agaacacata
      421 gcgacatgca aatattaaaa aacggaaacc ccgggaatct aacccggctg aacggattta
      481 gagtccgttc gatctacatg atcaggtttc cggtgtttcg tcctttccac aagatatata
      541 aagccaagaa atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac
      601 ataattttaa aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta
      661 ctaatatctt tgtgtttaca gtcaaattaa ttctaattat ctctctaaca gccttgtatc
      721 gtatatgcaa atatgaagga atcatgggaa ataggccctc ttcctgcccg acctaggctc
      781 aagcagtgat cagatccaga catgataaga tacattgatg agtttggaca aaccacaact
      841 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
      901 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
      961 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     1021 gctgattatg atcctctagt acttctcgac aagcttggcc ggcctggcct tatcactttc
     1081 caagtcggtt catctctatg tctgtataaa tctgtctttt cttggtgtgc tttaatttaa
     1141 tgcaaagatg gataccaact cggagaacca agaatagtcc aatgattaac cctatgataa
     1201 agaaaaaaga ggcaatagag cttttccaac tactgaacca accttctaca agctcgattg
     1261 gatttttgga tagcccagta tcaccaaaaa ataaactctc atcatcagga agttgcgaag
     1321 cagcgtcttg aatgtgagga tgttcgaaca cctgagcctt tgagctaaga tgaagatcgg
     1381 agtccaacat accatgtcca atcatgtata aaggaaactt atatcctgaa ctggtcctca
     1441 gaactccatt gggtccaatt tccacgtctt catatggtgc ccagtcatcc cacagttccc
     1501 tttctgtggt agttccactg atcattccga ccattcttga gaggattgga gcagcaatat
     1561 cgactctgat gtatctggtc tcaaagtatt ttagggtacc attgattatg gtgaaagcag
     1621 gaccggttcc tgggttttta ggagcaagat agctgagatc cactggagag attggaagac
     1681 ccgctctgat tttgctccag gtttcttggc agagggaata atccaagatc ctctcaacgt
     1741 cctgaattag acttacatcc actgaggtct gagatggagc agagatactt gacccttctg
     1801 ggcattcagg gaatctggct gcagcaaaga gatccttatc agccatctcg aaccagacac
     1861 ctgatgggag tctgactccc caatgcttgc agtattgcat tttgcaggcc ttgcctccag
     1921 tttcataagc aaagtagtta cttctgaacc ctgtgccctc ctttcccagg gatgatagct
     1981 ctccgtcctc tgagaagaag gtgatgtcca tggaaatgag gttagaatca catagccctt
     2041 tgaccttata gtcagaatgc caggttgtag agttatggac agtggggcat atgtaattgc
     2101 tgcattttcc gttgatgaac tgtgaatcaa cccattctcc tgtgtattca tcaaccagca
     2161 catggtgagg agtcacctgg acaatcactg cttcggcatc cgtcacagtt gcatatccac
     2221 aactttgagg agggaagcct ggattcagcc aagttccttg tttcgtttgt tcaatgcttt
     2281 ccttgcattg ttctacagat ggagtgaagg atcggatgga atgtgttata tacttcggtc
     2341 cataccagcg gaaatcacaa gtagtgaccc atttggaagc atgacacatc caaccgtctg
     2401 cttgaatagc cttgtgactc ttgggcattt tgacttgtaa ggctgtgcct attaagtcat
     2461 tatgccaatt taaatctgag cttgacgggc aataatggta attagaagga acatttttcc
     2521 agtttccttt ttggttgtgt ggaaaaacta tggtgaactt gcaattcacc ccaatgaata
     2581 aaaaggctaa gtacaaaagg cacttcatgg tggcggtttt ttaggagttc gcgcccgatg
     2641 gtgggacggt atgaatcatc cggaatattt ataggttttt ttattacaaa actgttacga
     2701 aaacagtaaa atacttattt atttgcgaga tggttatcat tttaattatc tccatgatct
     2761 attaatattc cggagtatac ggaccgcggc cgcaaatacc tgcaggatcc gttttgcgct
     2821 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     2881 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     2941 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     3001 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     3061 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     3121 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     3181 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     3241 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     3301 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     3361 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     3421 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     3481 taatacgact cactataggg agacccaagc tggctagcgc cgccaccatg gtttctaaag
     3541 gtgaagaact ttttactggc gttgtgccga ttctggtgga actggatggt gatgtgaatg
     3601 gccataaatt tagcgttcgt ggcgaaggcg aaggtgatgc gaccaacggt aaactgaccc
     3661 tgaaatttat ttgcaccacc ggtaaactgc cggttccgtg gccgaccctg gtgaccaccc
     3721 tgacctatgg cgttcagtgc tttagccgct atccggatca tatgaaacgc catgatttct
     3781 ttaaaagcgc gatgccggaa ggctatgtgc aggaacgtac cattagcttc aaagatgatg
     3841 gcacctataa aacccgtgcg gaagttaaat ttgaaggcga taccctggtg aaccgcattg
     3901 aactgaaagg tattgatttt aaagaagatg gcaacattct gggtcataaa ctggaatata
     3961 atttcaacag ccataatgtg tatattaccg ccgataaaca gaaaaatggc atcaaagcga
     4021 actttaaaat ccgtcacaac gtggaagatg gtagcgtgca gctggcggat cattatcagc
     4081 agaatacccc gattggtgat ggcccggtgc tgctgccgga taatcattat ctgagcaccc
     4141 agagcgttct gagcaaagat ccgaatgaaa aacgtgatca tatggtgctg ctggaatttg
     4201 ttaccgccgc gggcattacc cacggtatgg atgaacttta taaaggttct ggaaagccga
     4261 ttccaaatcc cctgttggga ctggattcta ctcatcacca ccatcaccac taagaattca
     4321 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4381 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4441 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     4501 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     4561 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4621 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4681 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     4741 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     4801 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4861 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4921 ccgcgtcgac tttaactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     4981 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5041 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     5101 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     5161 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     5221 accagatctc gggcaggaag agggcctatt tcccatgatt ccttcatatt tgcatatacg
     5281 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
     5341 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     5401 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     5461 atatatcttg tggaaaggac gaaacaccgg aaacctgatc atgtagatcg aacggactct
     5521 aaatccgttc agccgggtta gattcccggg gtttccgttt tttaatattt gcatgtcgct
     5581 atgtgttctg ggaaatcacc ataaacgtga aatccctatc agtgatagag acttataagt
     5641 tccctatcag tgatagagac accggggggt ggatcgaata gatcacacgg actctaaatc
     5701 cgtgcaggcg ggtgaaactc ccgcaccccc cgttttttca gctggggctc tagggggtat
     5761 ccccacgcgc cctgtagcgg cgcattaagc gcaagccctg ccatagccac tacgggtacg
     5821 tagccaacca ctagaactat agctagagtc ctgggcgaac aaacgatgct cgccttccag
     5881 aaaaccgagg atgcgaacca cttcatccgg ggtcagcacc accggcaagc gccgcgacgg
     5941 ccgaggtcta ccgatctcct gaagcaaggg cagatccgtg cacagcacct tgccgtagaa
     6001 gaacagcaag gccgccaatg cctgacgatg cgtggagacc gaaaccttgc gctcgttcgc
     6061 cagccaggac agaaatgcct cgacttcgct gctgcccaag gttgccgggt gacgcacacc
     6121 gtggaaacgg atgaaggcac gaacccagtt gacataagcc tgttcggttc gtaaactgta
     6181 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     6241 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgtac agtctatgcc
     6301 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     6361 caacgatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     6421 taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat
     6481 ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc acctactccc
     6541 aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc
     6601 ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt
     6661 ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccggc gagcaccgga
     6721 ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg
     6781 cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa
     6841 agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac
     6901 aattcgttca agccgagatc ggcttcccgg ccgcggagtt gttcggtaaa ttgtcacaac
     6961 gccgcgaata tagtctttac catgcccttg gccacgcccc tctttaatac gacgggcaat
     7021 ttgcacttca gaaaatgaag agtttgcttt agccataaca aaagtccagt atgctttttc
     7081 acagcataac tggactgatt tcagtttaca actattctgt ctagtttaag actttattgt
     7141 catagtttag atctattttg ttcagtttaa gactttattg tccgcccaca cccgcttacg
     7201 cagggcatcc atttattact caaccgtaac cgattttgcc aggttacgcg gctggtctat
     7261 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
     7321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7921 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     8761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9241 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     9301 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     9361 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaag aacgtggact
     9421 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     9481 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     9541 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     9601 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     9661 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     9721 aaataagcgt tgatattcag tcaattacaa acattaataa cgaagagatg acagaaaaat
     9781 tttcattctg tgacagagaa
//
