LOCUS       sequence-216936-        9440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the drug-activated transcription factor rTetR-NS3-VP64-p65 
            along with IRES mCherry as a fluorescent marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9440)
  AUTHORS   Tague EP, Dotson HL, Tunney SN, Sloas DC, Ngo JT
  TITLE     Chemogenetic control of gene expression and cell signaling with 
            antiviral drugs.
  JOURNAL   Nat Methods. 2018 Jul;15(7):519-522. doi: 10.1038/s41592-018-0042-y.
            Epub 2018 Jul 2.
  PUBMED    29967495
REFERENCE   2  (bases 1 to 9440)
  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: 
            "10.1038/s41592-018-0042-y"; journalName: "Nat Methods"; date: 
            "2018-07"; volume: "15"; issue: "7"; pages: "519-522"
COMMENT     Sequence Label: rTetR-NS3-VP64-p65_IRES_mCherry
FEATURES             Location/Qualifiers
     source          1..9440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(476..1267,1268..1336))
                     /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     1099..1118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1337..1441)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1516..1535)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1707..2086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2087..2290
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2241..2261
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2335..2354
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2335..2353
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3859..3885
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3904..3924
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    3961..3985
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4015..4164
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             4189..4209
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4639..4995
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             5592..6302
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5737..5755)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6005..6024
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(6334..6351)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6340..6564
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6610..7038
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6697..6716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6907..6928
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7047..7067)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7052..7381
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7232..7367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7294..7313
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7430..8455
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             7430..8455
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    8585..8706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8622..8641)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8676..8695
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8755..8771)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8755..8771)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8768..8790)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8779..8795
                     /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(8803..8809,8810..8827,8828..8833))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8848..8869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8986..9003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(9157..9440,1..305))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9237..9256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
       61 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      121 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      181 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      241 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
      301 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
      361 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
      421 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
      481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      541 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      601 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
      661 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      721 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
      781 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      841 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      901 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      961 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1021 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1081 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1141 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1201 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1261 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1321 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1381 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1441 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     1501 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     1561 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     1621 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     1681 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     1741 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1801 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1861 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1921 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     1981 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2041 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2101 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2161 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2221 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2281 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     2341 gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttgc caccatgtct
     2401 agactggaca agagcaaagt cataaacgga gctctggaat tactcaatgg tgtcggtatc
     2461 gaaggcctga cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac
     2521 tggcacgtga agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg
     2581 catcataccc acttctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac
     2641 gccaagtcat accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc
     2701 acccgcccaa cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag
     2761 caaggcttct ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg
     2821 ggctgcgtat tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc
     2881 accgattcta tgcccccact tctgagacaa gcaattgagc tgttcgaccg gcagggagcc
     2941 gaacctgcct tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag
     3001 tgcgaaagcg gggcgtctgc aggcatggcc agcatgactg gtggacagca aatggggtcg
     3061 acggaggacg tggtgtgctg ccactcaatc tacggcaaga agaagggtga tatcgacacc
     3121 taccgataca taggctcttc cgggacaggc tgcgtggtca tagtgggcag gatcgtcttg
     3181 tccggatccg gcactagtgc gcccatcacg gcgtacgccc agcagacgag aggcctccta
     3241 gggtgtataa tcaccagcct gactggccgg gacaaaaacc aagtggaggg tgaggtccag
     3301 atcgtgtcaa ctgctaccca aaccttcctg gcaacgtgca tcaatggggt atgctgggca
     3361 gtctaccacg gggccggaac gaggaccatc gcatcaccca agggtcctgt catccagatg
     3421 tataccaatg tggaccaaga ccttgtgggc tggcccgctc ctcaaggttc ccgctcattg
     3481 acaccctgta cctgcggctc ctcggacctt tacctggtca cgaggcacgc cgatgtcatt
     3541 cccgtgcgcc ggcgaggtga tagcaggggt agcctgcttt cgccccggcc catttcctac
     3601 ttgaaaggct cctcgggggg tccgctgttg tgccccgcgg gacacgccgt gggcctattc
     3661 agggccgcgg tgtgcacccg tggagtggct aaagcggtgg actttatccc tgtggagaac
     3721 ctagagacaa ccatgagatc cccggtgttc acggacaact cctctccacc agcagtcacc
     3781 ctgacgcacc caatcaccaa aatcgatagg gaggttctct accaggagtt cgatgagatg
     3841 gaagagtgct ctcagcacta tccctacgat gtgcccgatt acgctggcgc gtcgtctgca
     3901 tgccccaaga agaagaggaa ggtgtcgcca gggatccgtc gacttgacgc gttgatatca
     3961 acaagtttgt acaaaaaagc aggctacaaa gaggccagcg gttccggacg ggctgacgca
     4021 ttggacgatt ttgatctgga tatgctggga agtgacgccc tcgatgattt tgaccttgac
     4081 atgcttggtt cggatgccct tgatgacttt gacctcgaca tgctcggcag tgacgccctt
     4141 gatgatttcg acctggacat gctgattaac tctagaagtt ccggatctcc gaaaaagaaa
     4201 cgcaaagttg gtagccagta cctgcccgac accgacgacc ggcaccggat cgaggaaaag
     4261 cggaagcgga cctacgagac attcaagagc atcatgaaga agtccccctt cagcggcccc
     4321 accgacccta gacctccacc tagaagaatc gccgtgccca gcagatccag cgccagcgtg
     4381 ccaaaacctg ccccccagcc ttaccccttc accagcagcc tgagcaccat caactacgac
     4441 gagttcccta ccatggtgtt ccccagcggc cagatctctc aggcctctgc tctggctcca
     4501 gcccctcctc aggtgctgcc tcaggctcct gctcctgcac cagctccagc catggtgtct
     4561 gcactggctc aggcaccagc acccgtgcct gtgctggctc ctggacctcc acaggctgtg
     4621 gctccaccag cccctaaacc tacacaggcc ggcgagggca cactgtctga agctctgctg
     4681 cagctgcagt tcgacgacga ggatctggga gccctgctgg gaaacagcac cgatcctgcc
     4741 gtgttcaccg acctggccag cgtggacaac agcgagttcc agcagctgct gaaccagggc
     4801 atccctgtgg cccctcacac caccgagccc atgctgatgg aataccccga ggccatcacc
     4861 cggctcgtga caggcgctca gaggcctcct gatccagctc ctgcccctct gggagcacca
     4921 ggcctgccta atggactgct gtctggcgac gaggacttca gctctatcgc cgatatggat
     4981 ttctcagcct tgctgtgatc tagagggccc ggtattgcgg tacccttgta cgcctgtttt
     5041 atactccctt cccgtaactt agacgcacaa aaccaagttc aatagaaggg ggtacaaacc
     5101 agtaccacca cgaacaagca cttctgtttc cccggtgatg tcgtatagac tgcttgcgtg
     5161 gttgaaagcg acggatccgt tatccgctta tgtacttcga gaagcccagt accacctcgg
     5221 aatcttcgat gcgttgcgct cagcactcaa ccccagagtg tagcttaggc tgatgagtct
     5281 ggacatccct caccggtgac ggtggtccag gctgcgttgg cggcctacct atggctaacg
     5341 ccatgggacg ctagttgtga acaaggtgtg aagagcctat tgagctacat aagaatcctc
     5401 cggcccctga atgcggctaa tcccaacctc ggagcaggtg gtcacaaacc agtgattggc
     5461 ctgtcgtaac gcgcaagtcc gtggcggaac cgactacttt gggtgtccgt gtttcctttt
     5521 attttattgt ggctgcttat ggtgacaatc acagattgtt atcataaagc gaattggata
     5581 ggatccgtac gatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     5641 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     5701 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     5761 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     5821 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     5881 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     5941 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     6001 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     6061 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     6121 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     6181 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     6241 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     6301 aatctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     6361 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     6421 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     6481 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     6541 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     6601 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     6661 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     6721 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     6781 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     6841 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     6901 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     6961 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     7021 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     7081 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     7141 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     7201 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     7261 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     7321 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     7381 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtga tgaaaaagcc
     7441 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     7501 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     7561 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     7621 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     7681 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     7741 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     7801 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     7861 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     7921 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     7981 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     8041 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     8101 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     8161 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     8221 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     8281 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     8341 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     8401 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     8461 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     8521 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     8581 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     8641 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     8701 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     8761 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     8821 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     8881 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     8941 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     9001 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     9061 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     9121 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     9181 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     9241 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     9301 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     9361 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     9421 ccccgttcag cccgaccgct
//
