LOCUS       sequence-91429-e        9030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli genome modification .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9030)
  AUTHORS   Yang J, Sun B, Huang H, Jiang Y, Diao L, Chen B, Xu C, Wang X, Liu 
            J, Jiang W, Yang S
  TITLE     High-Efficiency Scarless Genetic Modification in Escherichia coli by
            Using Lambda Red Recombination and I-SceI Cleavage.
  JOURNAL   Appl Environ Microbiol. 2014 Jul 1;80(13):3826-3834. Epub 2014 Apr 
            18.
  PUBMED    24747889
REFERENCE   2  (bases 1 to 9030)
  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: "Appl 
            Environ Microbiol."; date: "2014-07-1"; volume: "80(13)"; pages: 
            "3826-3834. Epub 2014 Apr 18"
COMMENT     Sequence Label: pREDIA
FEATURES             Location/Qualifiers
     source          1..9030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(11..889)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        916..1200
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1123..1142
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1227..1235
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1244..1660
                     /codon_start=1
                     /gene="gam"
                     /product="inhibitor of the host RecBCD nuclease in the 
                     lambda Red system"
                     /label=Gam
                     /note="protects the double-stranded DNA substrate for 
                     recombination"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             1666..2451
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     CDS             2448..3128
                     /codon_start=1
                     /gene="exo"
                     /product="5' to 3' double-stranded DNA exonuclease in the 
                     lambda Red system"
                     /label=Exo
                     /note="creates 3' single-stranded DNA tails"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     terminator      3129..3373
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     CDS             complement(3406..4254)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(4328..5164)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        5310..5428
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             complement(6197..7147)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      7195..7417
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(join(8039..8830,8831..8899))
                     /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     8662..8681
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8900..9004)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catcgattta ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
       61 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
      121 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
      181 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
      241 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
      301 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
      361 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
      421 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
      481 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
      541 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
      601 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
      661 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
      721 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
      781 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
      841 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
      901 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
      961 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
     1021 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
     1081 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
     1141 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
     1201 acccgttttt ttgggaattc gagctctaag gaggttataa aaaatggata ttaatactga
     1261 aactgagatc aagcaaaagc attcactaac cccctttcct gttttcctaa tcagcccggc
     1321 atttcgcggg cgatattttc acagctattt caggagttca gccatgaacg cttattacat
     1381 tcaggatcgt cttgaggctc agagctgggc gcgtcactac cagcagctcg cccgtgaaga
     1441 gaaagaggca gaactggcag acgacatgga aaaaggcctg ccccagcacc tgtttgaatc
     1501 gctatgcatc gatcatttgc aacgccacgg ggccagcaaa aaatccatta cccgtgcgtt
     1561 tgatgacgat gttgagtttc aggagcgcat ggcagaacac atccggtaca tggttgaaac
     1621 cattgctcac caccaggttg atattgattc agaggtataa aacgaatgag tactgcactc
     1681 gcaacgctgg ctgggaagct ggctgaacgt gtcggcatgg attctgtcga cccacaggaa
     1741 ctgatcacca ctcttcgcca gacggcattt aaaggtgatg ccagcgatgc gcagttcatc
     1801 gcattactga tcgttgccaa ccagtacggc cttaatccgt ggacgaaaga aatttacgcc
     1861 tttcctgata agcagaatgg catcgttccg gtggtgggcg ttgatggctg gtcccgcatc
     1921 atcaatgaaa accagcagtt tgatggcatg gactttgagc aggacaatga atcctgtaca
     1981 tgccggattt accgcaagga ccgtaatcat ccgatctgcg ttaccgaatg gatggatgaa
     2041 tgccgccgcg aaccattcaa aactcgcgaa ggcagagaaa tcacggggcc gtggcagtcg
     2101 catcccaaac ggatgttacg tcataaagcc atgattcagt gtgcccgtct ggccttcgga
     2161 tttgctggta tctatgacaa ggatgaagcc gagcgcattg tcgaaaatac tgcatacact
     2221 gcagaacgtc agccggaacg cgacatcact ccggttaacg atgaaaccat gcaggagatt
     2281 aacactctgc tgatcgccct ggataaaaca tgggatgacg acttattgcc gctctgttcc
     2341 cagatatttc gccgcgacat tcgtgcatcg tcagaactga cacaggccga agcagtaaaa
     2401 gctcttggat tcctgaaaca gaaagccgca gagcagaagg tggcagcatg acaccggaca
     2461 ttatcctgca gcgtaccggg atcgatgtga gagctgtcga acagggggat gatgcgtggc
     2521 acaaattacg gctcggcgtc atcaccgctt cagaagttca caacgtgata gcaaaacccc
     2581 gctccggaaa gaagtggcct gacatgaaaa tgtcctactt ccacaccctg cttgctgagg
     2641 tttgcaccgg tgtggctccg gaagttaacg ctaaagcact ggcctgggga aaacagtacg
     2701 agaacgacgc cagaaccctg tttgaattca cttccggcgt gaatgttact gaatccccga
     2761 tcatctatcg cgacgaaagt atgcgtaccg cctgctctcc cgatggttta tgcagtgacg
     2821 gcaacggcct tgaactgaaa tgcccgttta cctcccggga tttcatgaag ttccggctcg
     2881 gtggtttcga ggccataaag tcagcttaca tggcccaggt gcagtacagc atgtgggtga
     2941 cgcgaaaaaa tgcctggtac tttgccaact atgacccgcg tatgaagcgt gaaggcctgc
     3001 attatgtcgt gattgagcgg gatgaaaagt acatggcgag ttttgacgag atcgtgccgg
     3061 agttcatcga aaaaatggac gaggcactgg ctgaaattgg ttttgtattt ggggagcaat
     3121 ggcgatgacg catcctcacg ataatatccg ggtaggcgca atcactttcg tctactccgt
     3181 tacaaagcga ggctgggtat ttcccggcct ttctgttatc cgaaatccac tgaaagcaca
     3241 gcggctggct gaggagataa ataataaacg aggggctgta tgcacaaagc atcttctgtt
     3301 gagttaagaa cgagtatcga gatggcacat agccttgctc aaattggaat caggtttgtg
     3361 ccaataccag tagaaacaga cgaagaatcc atggggcatg gcgaattaat ctttctgcga
     3421 attgagatga cgccactggc tgggcgtcat cccggtttcc cgggtaaaca ccaccgaaaa
     3481 atagttacta tcttcaaagc cacattcggt cgaaatatca ctgattaaca ggcggctatg
     3541 ctggagaaga tattgcgcat gacacactct gacctgtcgc agatattgat tgatggtcat
     3601 tccagtctgc tggcgaaatt gctgacgcaa aacgcgctca ctgcacgatg cctcatcaca
     3661 aaatttatcc agcgcaaagg gacttttcag gctagccgcc agccgggtaa tcagcttatc
     3721 cagcaacgtt tcgctggatg ttggcggcaa cgaatcactg gtgtaacgat ggcgattcag
     3781 caacatcacc aactgcccga acagcaactc agccatttcg ttagcaaacg gcacatgctg
     3841 actactttca tgctcaagct gaccgataac ctgccgcgcc tgcgccatcc ccatgctacc
     3901 taagcgccag tgtggttgcc ctgcgctggc gttaaatccc ggaatcgccc cctgccagtc
     3961 aagattcagc ttcagacgct ccgggcaata aataatattc tgcaaaacca gatcgttaac
     4021 ggaagcgtag gagtgtttat cgtcagcatg aatgtaaaag agatcgccac gggtaatgcg
     4081 ataagggcga tcgttgagta catgcaggcc attaccgcgc cagacaatca ccagctcaca
     4141 aaaatcatgt gtatgttcag caaagacatc ttgcggataa cggtcagcca cagcgactgc
     4201 ctgctggtcg ctggcaaaaa aatcatcttt gagaagtttt aactgatgcg ccaccgtggc
     4261 tacctcggcc agagaacgaa gttgattatt cgcaatatgg cgtacaaata cgttgagaag
     4321 attcgcgtta ttgcagaaag ccatcccgtc cctggcgaat atcacgcggt gaccagttaa
     4381 actctcggcg aaaaagcgtc gaaaagtggt tactgtcgct gaatccacag cgataggcga
     4441 tgtcagtaac gctggcctcg ctgtggcgta gcagatgtcg ggctttcatc agtcgcaggc
     4501 ggttcaggta tcgctgaggc gtcagtcccg tttgctgctt aagctgccga tgtagcgtac
     4561 gcagtgaaag agaaaattga tccgccacgg catcccaatt cacctcatcg gcaaaatggt
     4621 cctccagcca ggccagaagc aagttgagac gtgatgcgct gttttccagg ttctcctgca
     4681 aactgctttt acgcagcaag agcagtaatt gcataaacaa gatctcgcga ctggcggtcg
     4741 agggtaaatc attttcccct tcctgctgtt ccatctgtgc aaccagctgt cgcacctgct
     4801 gcaatacgct gtggttaacg cgccagtgag acggatactg cccatccagc tcttgtggca
     4861 gcaactgatt cagcccggcg agaaactgaa atcgatccgg cgagcgatac agcacattgg
     4921 tcagacacag attatcggta tgttcataca gatgccgatc atgatcgcgt acgaaacaga
     4981 ccgtgccacc ggtgatggta tagggctgcc cattaaacac atgaataccc gtgccatgtt
     5041 cgacaatcac aatttcatga aaatcatgat gatgttcagg aaaatccgcc tgcgggagcc
     5101 ggggttctat cgccacggac gcgttaccag acggaaaaaa atccacacta tgtaatacgg
     5161 tcatactggc ctcctgatgt cgtcaacacg gcgaaatagt aatcacgagg tcaggttctt
     5221 accttaaatt ttcgacggaa aaccacgtaa aaaacgtcga tttttcaaga tacagcgtga
     5281 attttcagga aatgcggtga gcatcacatc accacaattc agcaaattgt gaacatcatc
     5341 acgttcatct ttccctggtt gccaatggcc cattttcctg tcagtaacga gaaggtcgcg
     5401 aattcaggcg ctttttagac tggtcgtaat gaaattcagc aggatcacat atgcatcaaa
     5461 aaaaccaggt aatgaacctg ggtccgaact ctaaactgct gaaagaatac aaatcccagc
     5521 tgatcgaact gaacatcgaa cagttcgaag caggtatcgg tctgatcctg ggtgatgctt
     5581 acatccgttc tcgtgatgaa ggtaaaacct actgtatgca gttcgagtgg aaaaacaaag
     5641 catacatgga ccacgtatgt ctgctgtacg atcagtgggt actgtctttc aaacaccaag
     5701 ctttcaacaa actggctaac ctgttcatcg ttaacaacaa aaaaaccatc ccgaacaacc
     5761 tggttgaaaa ctacctgacc ccgatgtctc tggcatactg gttcatggat gatggtggta
     5821 aatgggatta caacaaaaac tctaccaaca aatcgatcgt actgaacacc cagtctttca
     5881 ctttcgaaga agtagaatac ctggttaagg gtctgcgtaa caaattccaa ctgaactgtt
     5941 acgtaaaaat caacaaaaac aaaccgatca tctacatcga ttctatgtct tacctgatct
     6001 tctacaacct gatcaaaccg tacctgatcc cgcagatgat gtacaaactg ccgaacacta
     6061 tctcctccga aactttcctg aaataataac catgggtatg gacagttttc cctttgatat
     6121 gtaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac
     6181 aagagccata agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc
     6241 gttgtttttg cgtgagccat gagaacgaac cattgagatc atacttactt tgcatgtcac
     6301 tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg
     6361 tttttcttag tccgttacgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac
     6421 cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt
     6481 caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat
     6541 tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa
     6601 actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta
     6661 tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca
     6721 tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta
     6781 actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga
     6841 acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt
     6901 ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc
     6961 tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc
     7021 ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt
     7081 catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt
     7141 cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta
     7201 gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga
     7261 cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt
     7321 gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa
     7381 agataaaaag aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt
     7441 attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta
     7501 aaggcttaag tagcaccctc gcaagctcgg ttgcggccgc aatcgggcaa atcgctgaat
     7561 attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt
     7621 cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg
     7681 gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg
     7741 ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc
     7801 ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta
     7861 atgcacccag taaggcagcg gtatcatcaa cggggtctga cgctcagtgg aacgaaaact
     7921 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7981 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8041 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8101 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8161 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8221 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8281 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8341 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8401 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8461 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8521 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8581 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8641 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8701 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8761 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     8821 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     8881 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     8941 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     9001 cgcgcacatt tccccgaaaa gtgccacctg
//
