LOCUS       sequence-309439-        8895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid for insertion of eGFP and the constitutive acpP 
            promoter into the S. Typhimurium chromosome. GFP is in the forward 
            orientation relative to the kanamycin resistance selectable marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8895)
  AUTHORS   Holden ER, Wickham GJ, Webber MA, Thomson NM, Trampari E
  TITLE     Donor plasmids for phenotypically neutral chromosomal gene 
            insertions in Enterobacteriaceae.
  JOURNAL   Microbiology (Reading). 2020 Nov 23. doi: 10.1099/mic.0.000994.
  PUBMED    33226934
REFERENCE   2  (bases 1 to 8895)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Microbiology (Reading). 2020 Nov 23. doi: 10.1099/mic.0.000994."
COMMENT     Sequence Label: pDOC-K-glmS-GFPfor
FEATURES             Location/Qualifiers
     source          1..8895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1720..1739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1973..1990
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2107..2128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2143..2148,2149..2166,2167..2173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2181..2197
                     /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)."
     primer_bind     2186..2208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2205..2221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2205..2221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(2724..2771)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2981..3775
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3035..3054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3645..3664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    complement(3966..3999)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             4123..4839
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(4153..4181)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4776..4796
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5319..5336)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5319..5335)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5328..5350)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5515..5532)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5685..5771
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5863..5890
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     oriT            6337..6446
                     /note="incP origin of transfer"
     promoter        6820..7265
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(7266..7352,7353..8687)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     primer_bind     complement(8726..8745)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8845..8867
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta gccaccactt caagaactct gtagcaccgc ctacatacct
     1441 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1501 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1561 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1621 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1681 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     1741 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     1801 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     1861 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     1921 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     1981 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2041 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2101 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2161 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2221 ccatgattac gccaagctct agggataaca gggtaatcga tgaattcgct cgaaggcgcg
     2281 ctgaagctga aagagatctc ttacattcac gcggaagcct atgctgcagg cgagctgaaa
     2341 cacgggccgt tggcgctgat tgacgccgat atgccggtga ttgtggtggc gccgaacaac
     2401 gaactgctgg agaaactgaa atccaacatc gaagaagttc gcgcacgtgg cggtcagttg
     2461 tacgtcttcg ccgatcagga tgcgggtttc gtcagcaacg acaacatgca catcatcgag
     2521 atgccgcatg tggaagaggt cattgcgcct atcttctaca ccgttccgct gcaactgctg
     2581 gcgtatcacg tggcgctgat taaaggcacc gacgttgacc agccgcgtaa cctggcgaaa
     2641 tcggttaccg tagagtaagt acaaatgtag taccaggcag aaggccgggt acctaggacc
     2701 ggtcaattgg ctggagctgc ttcgaagttc ctatactttc tagagaatag gaacttcgga
     2761 ataggaactt caagatcccc cacgctgccg caagcactca gggcgcaagg gctgctaaag
     2821 gaagcggaac acgtagaact taagggaatt gccagctggg gcgccctctg gtaaggttgg
     2881 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     2941 atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3001 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3061 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3121 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3181 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3241 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3301 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3361 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3421 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3481 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3541 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3601 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3661 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgctct acggtatcgc
     3721 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3781 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3841 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3901 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccag cttcaaaagc
     3961 gctctgaagt tcctatactt tctagagaat aggaacttcg gaataggaac taaggaggat
     4021 attcatatct cgagcatatg cccgttgcaa atttttcaac attttataca ctacgaaaac
     4081 catcgcgaaa gcgagttttg gatttaagaa ggagatatac atatgagtaa aggagaagaa
     4141 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
     4201 ttttctgtca gtggagaggg tgaaggtgat gctacatacg gaaagcttac ccttaaattt
     4261 atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac tttgacctat
     4321 ggtgttcaat gcttttcccg ttatccggat catatgaaac ggcatgactt tttcaagagt
     4381 gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga cgggaactac
     4441 aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaaa
     4501 ggtattgatt ttaaagaaga tggaaacatt ctcggacaca aactcgagta caactataac
     4561 tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa
     4621 attcgccaca acattgaaga tggatccgtt caactagcag accattatca acaaaatact
     4681 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgcc
     4741 ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct
     4801 gctgggatta cacatggcat ggatgagctc tacaaataag gggcggccgc aagctttcga
     4861 cagacggcct ttttttgtgc gctgtgacag gcgctgttct ttctctatgg tggctgaaga
     4921 gcgactaaga ggtgctacgc ctgcctgcgc tacgggcacc cggtcacatt ctcttcttta
     4981 aggccctggc tcaatccagg tggatcggaa atcaaaccac cccaggttat tcaggtggat
     5041 ccccctggcc cggtcggggc ttttgagtcg catccagtgg tgaaaaagcg gctgtagcca
     5101 gcctgaacgg gtggtttccc tgaccagttg ttccgcgctg atggtttcgg catgccactg
     5161 ggtttcccat tgggccagca gcgcatcatc cccaccactg atggcgtggc gcagtaaagg
     5221 ggagcccagt agccatgtac cgacgttcca ctcttcggga atggggaaat tgaccggcta
     5281 gcgtcgacta gtagggataa cagggtaatg agcttggcac tggccgtcgt tttacaacgt
     5341 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5401 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5461 ctgaatggcg aatggcgagc ttggctgttt tggcggatga gagaagattt tcagcctgat
     5521 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
     5581 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
     5641 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     5701 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     5761 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     5821 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
     5881 atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg
     5941 catgcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatc
     6001 gacatccgcc ctcaccgcca ggaacgcaac cgcagcctca tcacgccggc gcttcttggc
     6061 cgcgcgggat tcaacccact cggccagctc gtcggtgtag ctctttggca tcgtctctcg
     6121 cctgtcccct cagttcagta atttcctgca tttgcctgtt tccagtcggt agatattcca
     6181 caaaacagca gggaagcagc gcttttccgc tgcataaccc tgcttcgggg tcattatagc
     6241 gattttttcg gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg
     6301 tgtagacttt ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc
     6361 acccgcgagc gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg
     6421 gaatcctgct ctgcgaggct ggccggctac cgccggcgta acagatgagg gcaagcggat
     6481 ggctgatgaa accaagccaa ccaggaaggg cagcccacct atcaaggtgt actgccttcc
     6541 agacgaacga agagcgattg aggaaaaggc ggcggcggcc ggcatgagcc tgtcggccta
     6601 cctgctggcc gtcggccagg gctacaaaat cacgggcgtc gtggactatg agcacgtccg
     6661 cgagctggcc cgcatcaatg gcgacctggg ccgcctgggc ggcctgctga aactctggct
     6721 caccgacgac ccgcgcacgg cgcggttcgg tgatgccacg atcctcgccc tgctggcgaa
     6781 gatcgactct agctagagga tcgatccttt ttaacccatc acatatacct gccgttcact
     6841 attatttagt gaaatgagat attatgatat tttctgaatt gtgattaaaa aggcaacttt
     6901 atgcccatgc aacagaaact ataaaaaata cagagaatga aaagaaacag atagattttt
     6961 tagttcttta ggcccgtagt ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga
     7021 aaatagacca gttgcaatcc aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc
     7081 gcgggtttgt tactgataaa gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac
     7141 tttggcgtca ccccttacat attttaggtc tttttttatt gtgcgtaact aacttgccat
     7201 cttcaaacag gagggctgga agaagcagac cgctaacaca gtacataaaa aaggagacat
     7261 gaacgatgaa catcaaaaag tttgcaaaac aagcaacagt attaaccttt actaccgcac
     7321 tgctggcagg aggcgcaact caagcgtttg cgaaagaaac gaaccaaaag ccatataagg
     7381 aaacatacgg catttcccat attacacgcc atgatatgct gcaaatccct gaacagcaaa
     7441 aaaatgaaaa atatcaagtt cctgagttcg attcgtccac aattaaaaat atctcttctg
     7501 caaaaggcct ggacgtttgg gacagctggc cattacaaaa cgctgacggc actgtcgcaa
     7561 actatcacgg ctaccacatc gtctttgcat tagccggaga tcctaaaaat gcggatgaca
     7621 catcgattta catgttctat caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg
     7681 ctggccgcgt ctttaaagac agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc
     7741 aaacacaaga atggtcaggt tcagccacat ttacatctga cggaaaaatc cgtttattct
     7801 acactgattt ctccggtaaa cattacggca aacaaacact gacaactgca caagttaacg
     7861 tatcagcatc agacagctct ttgaacatca acggtgtaga ggattataaa tcaatctttg
     7921 acggtgacgg aaaaacgtat caaaatgtac agcagttcat cgatgaaggc aactacagct
     7981 caggcgacaa ccatacgctg agagatcctc actacgtaga agataaaggc cacaaatact
     8041 tagtatttga agcaaacact ggaactgaag atggctacca aggcgaagaa tctttattta
     8101 acaaagcata ctatggcaaa agcacatcat tcttccgtca agaaagtcaa aaacttctgc
     8161 aaagcgataa aaaacgcacg gctgagttag caaacggcgc tctcggtatg attgagctaa
     8221 acgatgatta cacactgaaa aaagtgatga aaccgctgat tgcatctaac acagtaacag
     8281 atgaaattga acgcgcgaac gtctttaaaa tgaacggcaa atggtatctg ttcactgact
     8341 cccgcggatc aaaaatgacg attgacggca ttacgtctaa cgatatttac atgcttggtt
     8401 atgtttctaa ttctttaact ggcccataca agccgctgaa caaaactggc cttgtgttaa
     8461 aaatggatct tgatcctaac gatgtaacct ttacttactc acacttcgct gtacctcaag
     8521 cgaaaggaaa caatgtcgtg attacaagct atatgacaaa cagaggattc tacgcagaca
     8581 aacaatcaac gtttgcgcct agcttcctgc tgaacatcaa aggcaagaaa acatctgttg
     8641 tcaaagacag catccttgaa caaggacaat taacagttaa caaataaaaa cgcaaaagaa
     8701 aatgccgatt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     8761 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     8821 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     8881 caccgaaacg cgcga
//
