LOCUS       sequence-337785-       10350 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of orthogonal Mb tRNA and tRNA synthetase for 
            incorporation of tetrazine3.0 in eukaryotic cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10350)
  AUTHORS   Jang HS, Jana S, Blizzard RJ, Meeuwsen JC, Mehl RA
  TITLE     Access to Faster Eukaryotic Cell Labeling with Encoded Tetrazine 
            Amino Acids.
  JOURNAL   J Am Chem Soc. 2020 Apr 22;142(16):7245-7249. doi: 
            10.1021/jacs.9b11520. Epub 2020 Apr 10.
  PUBMED    32251579
REFERENCE   2  (bases 1 to 10350)
  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/jacs.9b11520"; journalName: "J Am Chem Soc"; date: 
            "2020-04-22- 22"; volume: "142"; issue: "16"; pages: "7245-7249"
COMMENT     Sequence Label: pAcBac1-NES-Flag-R2-84-MbRS Tetrazine3.0
FEATURES             Location/Qualifiers
     source          1..10350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..557
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     399..420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        584..688
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(689..757,758..1549)
                     /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(907..926)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1720..2308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2209..2228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2462..2479
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        complement(4143..4237)
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    4143..4161
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4171..4189
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(4314..4562)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     polyA_signal    4599..4823
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4812..4829
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    4878..5466
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4984..4995
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5393..5413
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6749..6772)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(6776..6802)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     regulatory      6823..6832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(6851..6870)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6852..6870)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6915..7118)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6944..6964)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        7119..7498
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7593..7684
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     7609..7626
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(7743..7790,7791..9278)
                     /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"
     polyA_signal    9424..9558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9461..9480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9515..9534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        9591..9839
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        9916..10010
                     /label=H1-2O2 promoter
                     /note="doxycycline-inducible variant of the human H1 RNA 
                     promoter (Henriksen et al., 2007)"
     protein_bind    9964..9982
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    9992..10010
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     mobile_element  10103..10268
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      421 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      481 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      541 attaacgttt acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      661 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      721 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      781 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
      841 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
      901 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
      961 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1021 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1081 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1141 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1201 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1261 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1321 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1381 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1441 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1501 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1561 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1621 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1681 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1741 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1801 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1861 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1921 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1981 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2041 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2101 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2161 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2221 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2281 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2341 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2401 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2461 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     2521 tctgtgcggt atttcacacc gcagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acctgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg ccctggcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcct acgtacccgt agtggctatg gcagggcttg cgcttaatgc
     4021 gccgctacag ggcgcgtggg gataccccct agagccccag ctgaaaaaac ggggggtgcg
     4081 ggagtttcac ccgcctgcac ggatttagag tccgtgtgat ctattcgatc caccccccgg
     4141 tgtctctatc actgataggg aacttataag tctctatcac tgatagggat ttcacgttta
     4201 tggtgatttc ccagaacaca tagcgacatg caaatattaa aaaacggaaa ccccgggaat
     4261 ctaacccggc tgaacggatt tagagtccgt tcgatctaca tgatcaggtt tccggtgttt
     4321 cgtcctttcc acaagatata taaagccaag aaatcgaaat actttcaagt tacggtaagc
     4381 atatgatagt ccattttaaa acataatttt aaaactgcaa actacccaag aaattattac
     4441 tttctacgtc acgtattttg tactaatatc tttgtgttta cagtcaaatt aattctaatt
     4501 atctctctaa cagccttgta tcgtatatgc aaatatgaag gaatcatggg aaataggccc
     4561 tcttcctgcc cgagatctgg ttctttccgc ctcagaagcc atagagccca ccgcatcccc
     4621 agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
     4681 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
     4741 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
     4801 tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg gtttaaacgg gccctctaga
     4861 ctcgagttaa agtcgacgcg gggaggcggc ccaaagggag atccgactcg tctgagggcg
     4921 aaggcgaaga cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc
     4981 tggattgagg gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac
     5041 acaggcgagc agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt
     5101 gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt
     5161 ggcaataggg agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc
     5221 ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac
     5281 gggccacaac tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa
     5341 agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt
     5401 aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt aatccagagg
     5461 ttgattgtcg acttaacgcg ttgaattctc atcacaggtt ggtgctgatg ccgttgtagt
     5521 agctctcgct tctgctggcc cgcttgatgt tcttgaagcc gtgcatcact ttcagcagcc
     5581 gttccaggcc gaagccggct ccgatccagg gcttgtcgat gccccactcc cggtccaggg
     5641 acacgggtcc caccacggcg ctgctcagtt ccaggtcgcc gtgcatgatg tccagggtgt
     5701 cgccgtacac catgcagctg tcgcccacga tctcgaagtc gatttccagg taatccagga
     5761 actctttgat cagggcttcc aggttctccc gggtgcagcc gctgcccatc tgggcaaagc
     5821 ccaccattgt aaactcttcc aggtgctctt tgccgtcgct ctctttccgg tagcagggtc
     5881 ccacttcgaa aatcttgatg gggccaggca ggattctgtc cagtttccgc aggtagttgt
     5941 agccggtggg ggccagcata ggccgcaggc acaggttctt gtccacccgg aaaatctgct
     6001 tggacagctc ggtgtcgttg ttgatgccca tccgctccac gtactcggcg gggatcagga
     6061 tggggctctt gatttccagg aagccccggt ccacgaagaa cttggtgatg tcccgttcca
     6121 gcttgcccag gtagtcctcc cggtcgttgg tgtacagccg ctggaagtcg ttcttccgcc
     6181 gggtcaccag ctcgggttcc agctcccgga agggcttggc catgttcagg ctgatcttgt
     6241 cctcggggga cagcagggcc tccactctgt ccagctggga ccgtgtcagg ctgggagcag
     6301 gagcagaggc gggcacggag ctgttggggg tgctcttggc ggggctgggc acgctgcggc
     6361 tggtgttggt gctggccttg gcggacacgc tgttttccag gggcttgggg gctctggaca
     6421 cgctcttggg catggctttc ttcactttgg gggcggacac cacccgcact ttcacgctgt
     6481 tcttgctctc ggtgcttctg gtcagaaagt tgttgatgtc ctcgtcggac acccggcacc
     6541 gcttgcaggt ctttctgtac ttgtggtgcc ggaaggctct ggcggtccgg cagcttctgc
     6601 tgttgttcac caccaggtgg tcgccgcagg ccatctcgat gtagattttg cttcttgaca
     6661 cctcgtggtg cttgatcttg tgcagggtgc cggtccggct catccacagg ccggtggcgc
     6721 tgatcagcac gtccaggggt ttcttgtcct tgtcgtcgtc gtccttgtag tcgtcgagtg
     6781 tgaggcgctc cagtggaggc aactgcaaag gaaccggaca cgccatggtg gcgctagcca
     6841 gcttgggtct ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct
     6901 ctagttagcc agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt
     6961 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
     7021 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
     7081 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
     7141 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
     7201 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
     7261 tgtactgcca agtgggcagt ttaccgtaaa tagtccaccc attgacgtca atggaaagtc
     7321 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
     7381 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
     7441 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa
     7501 cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac ggatcctgca ggtatttgcg
     7561 gccgcggtcc gtatactccg gaatattaat agatcatgga gataattaaa atgataacca
     7621 tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa aaaaacctat
     7681 aaatattccg gattattcat accgtcccac catcgggcgc gaactcctaa aaaaccgcca
     7741 ccatgaagtg ccttttgtac ttagcctttt tattcattgg ggtgaattgc aagttcacca
     7801 tagtttttcc acacaaccaa aaaggaaact ggaaaaatgt tccttctaat taccattatt
     7861 gcccgtcaag ctcagattta aattggcata atgacttaat aggcacagcc ttacaagtca
     7921 aaatgcccaa gagtcacaag gctattcaag cagacggttg gatgtgtcat gcttccaaat
     7981 gggtcactac ttgtgatttc cgctggtatg gaccgaagta tataacacat tccatccgat
     8041 ccttcactcc atctgtagaa caatgcaagg aaagcattga acaaacgaaa caaggaactt
     8101 ggctgaatcc aggcttccct cctcaaagtt gtggatatgc aactgtgacg gatgccgaag
     8161 cagtgattgt ccaggtgact cctcaccatg tgctggttga tgaatacaca ggagaatggg
     8221 ttgattcaca gttcatcaac ggaaaatgca gcaattacat atgccccact gtccataact
     8281 ctacaacctg gcattctgac tataaggtca aagggctatg tgattctaac ctcatttcca
     8341 tggacatcac cttcttctca gaggacggag agctatcatc cctgggaaag gagggcacag
     8401 ggttcagaag taactacttt gcttatgaaa ctggaggcaa ggcctgcaaa atgcaatact
     8461 gcaagcattg gggagtcaga ctcccatcag gtgtctggtt cgagatggct gataaggatc
     8521 tctttgctgc agccagattc cctgaatgcc cagaagggtc aagtatctct gctccatctc
     8581 agacctcagt ggatgtaagt ctaattcagg acgttgagag gatcttggat tattccctct
     8641 gccaagaaac ctggagcaaa atcagagcgg gtcttccaat ctctccagtg gatctcagct
     8701 atcttgctcc taaaaaccca ggaaccggtc ctgctttcac cataatcaat ggtaccctaa
     8761 aatactttga gaccagatac atcagagtcg atattgctgc tccaatcctc tcaagaatgg
     8821 tcggaatgat cagtggaact accacagaaa gggaactgtg ggatgactgg gcaccatatg
     8881 aagacgtgga aattggaccc aatggagttc tgaggaccag ttcaggatat aagtttcctt
     8941 tatacatgat tggacatggt atgttggact ccgatcttca tcttagctca aaggctcagg
     9001 tgttcgaaca tcctcacatt caagacgctg cttcgcaact tcctgatgat gagagtttat
     9061 tttttggtga tactgggcta tccaaaaatc caatcgagct tgtagaaggt tggttcagta
     9121 gttggaaaag ctctattgcc tcttttttct ttatcatagg gttaatcatt ggactattct
     9181 tggttctccg agttggtatc catctttgca ttaaattaaa gcacaccaag aaaagacaga
     9241 tttatacaga catagagatg aaccgacttg gaaagtgata aggccaggcc ggccaagctt
     9301 gtcgagaagt actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     9361 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     9421 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     9481 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     9541 tcttatcatg tctggatctg atcactgctt gagcctaggt cgggcaggaa gagggcctat
     9601 ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag ataattagaa
     9661 ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat
     9721 ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg
     9781 taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga cgaaacaccg
     9841 gaaacctgat catgtagatc gaacggactc taaatccgtt cagccgggtt agattcccgg
     9901 ggtttccgtt ttttaatatt tgcatgtcgc tatgtgttct gggaaatcac cataaacgtg
     9961 aaatccctat cagtgataga gacttataag ttccctatca gtgatagaga caccgggggg
    10021 tggatcgaat agatcacacg gactctaaat ccgtgcaggc gggtgaaact cccgcacccc
    10081 ccgttttttg ctaggagatc cgaaccagat aagtgaaatc tagttccaaa ctattttgtc
    10141 atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat tttgtcactc
    10201 ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa ctattttgtc
    10261 cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc tgccaactcc
    10321 atgtgacaaa ccgtcatctt cggctacttt
//
