LOCUS       sequence-215173-        8849 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Piggybac transposon plasmid CRISPR gene deletion activatable 
            fluorescence. Constitutive iRFP670 under EF1A promoter, CMV promoter
            with two SV40 polyA followed by red-enhanced nanolantern (ReNL).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8849)
  AUTHORS   Rui Y, Wilson DR, Choi J, Varanasi M, Sanders K, Karlsson J, Lim M, 
            Green JJ
  TITLE     Carboxylated branched poly(beta-amino ester) nanoparticles enable 
            robust cytosolic protein delivery and CRISPR-Cas9 gene editing.
  JOURNAL   Sci Adv. 2019 Dec 6;5(12):eaay3255. doi: 10.1126/sciadv.aay3255. 
            eCollection 2019 Dec.
  PUBMED    31840076
REFERENCE   2  (bases 1 to 8849)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2019-12-6"; volume: "5(12):eaay3255. doi"; pages: " 
            10.1126/sciadv.aay3255. eCollection 2019 Dec"
COMMENT     Sequence Label: PB-iRFP-STOP-ReNL
FEATURES             Location/Qualifiers
     source          1..8849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..118
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    415..448
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        507..718
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             731..999
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1062..1082
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1119..1128
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1125..2060
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     polyA_signal    2086..2197
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        2361..2664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2665..2868
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2819..2839
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2890..2923
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3090..3224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3344..3478
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    3507..3540
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5552..5571)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5617..5672
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5671..5690)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     repeat_region   6340..6374
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6397..6413)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6397..6413)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6410..6432)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6421..6437
                     /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(6445..6451,6452..6469,6470..6475))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6490..6511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6570..6765
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6616..6751
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6771..6905
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6973..6990)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7144..7732)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7224..7243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7903..8694,8695..8763))
                     /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     8526..8545
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8764..8849,1..19))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctggtcga cttaacccta
       61 gaaagataat catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg
      121 ttttatcgat ctgtatatcg aggtttattt attaatttga atagatatta agttttatta
      181 tatttacact tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt
      241 attaaaaaaa aacaaaaact caaaatttct tctataaagt aacaaaactt ttaaacattc
      301 tctcttttac aaaaataaac ttattttgta ctttaaaaac agtcatgttg tattataaaa
      361 taagtaatta gcttaactta tacataatag aaacaaatta tacttattaa tcgcataact
      421 tcgtataatg tatgctatac gaagttatgc gcagtagtcc aactagtctc gtgcaaggat
      481 ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
      541 gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg
      601 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
      661 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct
      721 gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct gaggccgcca
      781 tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct gaactgcgtc
      841 cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg cgctcccttg
      901 gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg ctcaactcta
      961 cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac cggcgcctac
     1021 ctccttcccc tccggccgca agtgaaaaca tccctatggc tccaaagaag aagcgtaagg
     1081 tacggtaccc agctttctac aaagtggcca ccccggtcgc caccatggcg cgtaaggtcg
     1141 atctcacctc ctgcgatcgc gagccgatcc acatccccgg cagcattcag ccgtgcggct
     1201 gcctgctagc ctgcgacgcg caggcggtgc ggatcacgcg cattacggaa aatgccggcg
     1261 cgttctttgg acgcgaaact ccgcgggtcg gtgagctact cgccgattac ttcggcgaga
     1321 ccgaagccca tgcgctgcgc aacgcactgg cgcagtcctc cgatccaaag cgaccggcgc
     1381 tgatcttcgg ttggcgcgac ggcctgaccg gccgcacctt cgacatctca ctgcatcgcc
     1441 atgacggtac atcgatcatc gagttcgagc ctgcggcggc cgaacaggcc gacaatccgc
     1501 tgcggctgac gcggcagatc atcgcgcgca ccaaagaact gaagtcgctc gaagagatgg
     1561 ccgcacgggt gccgcgctat ctgcaggcga tgctcggcta tcaccgcgtg atgttgtacc
     1621 gcttcgcgga cgacggctcc gggatggtga tcggcgaggc gaagcgcagc gacctcgaga
     1681 gctttctcgg tcagcacttt ccggcgtcgc tggtcccgca gcaggcgcgg ctactgtact
     1741 tgaagaacgc gatccgcgtg gtctcggatt cgcgcggcat cagcagccgg atcgtgcccg
     1801 agcacgacgc ctccggcgcc gcgctcgatc tgtcgttcgc gcacctgcgc agcatctcgc
     1861 cctgccatct cgaatttctg cggaacatgg gcgtcagcgc ctcgatgtcg ctgtcgatca
     1921 tcattgacgg cacgctatgg ggattgatca tctgtcatca ttacgagccg cgtgccgtgc
     1981 cgatggcgca gcgcgtcgcg gccgaaatgt tcgccgactt cttatcgctg cacttcaccg
     2041 ccgcccacca ccaacgctaa gcgtgttgca cttaacgcgt accgactgtg ccttctagtt
     2101 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2161 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2221 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacactcctt
     2281 cccctccggc cgcaagtgaa aacatcccta tagtaatcaa ttacggggtc attagttcat
     2341 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     2401 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2461 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     2521 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     2581 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     2641 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2701 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2761 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2821 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
     2881 cgtcagatca taacctcgta taatgtatgc tatacgaagt tattcgcgat gaataaatga
     2941 aagcttgcag atctgcgact ctagaggatc tgcgactcta gaggatcata atcagccata
     3001 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     3061 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     3121 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3181 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgcgac tctagaggat
     3241 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3301 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3361 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3421 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatccc
     3481 catcaagctg atccggaacc cttaatataa cctcgtataa tgtatgctat acgaagttat
     3541 tctagagcta gcgaattcat ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc
     3601 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     3661 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     3721 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     3781 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     3841 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     3901 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     3961 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     4021 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     4081 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     4141 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     4201 tacgagcgct ccgagggccg ccaccacctg ttcctggggc atggcaccgg cagcaccggc
     4261 agcggcagct ccggcaccgc ctcctccgag gacaacaaca tggccgtcat caaagagttc
     4321 atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc
     4381 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     4441 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg
     4501 tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc
     4561 aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc
     4621 tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc
     4681 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
     4741 ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc
     4801 cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc
     4861 tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg
     4921 gaacagtacg agcgctccga gggccgccac cacctgttcc ggctggaaga tttcgttggg
     4981 gactggcgac agacagccgg ctacaacctg gaccaagtcc ttgaacaggg aggtgtgtcc
     5041 agtttgtttc agaatctcgg ggtgtccgta actccgatcc aaaggattgt cctgagcggt
     5101 gaaaatgggc tgaagatcga catccatgtc atcatcccgt atgaaggtct gagcggcgac
     5161 caaatgggcc agatcgaaaa aatttttaag gtggtgtacc ctgtggatga tcatcacttt
     5221 aaggtgatcc tgcactatgg cacactggta atcgacgggg ttacgccgaa catgatcgac
     5281 tatttcggac ggccgtatga aggcatcgcc gtgttcgacg gcaaaaagat cactgtaaca
     5341 gggaccctgt ggaacggcaa caaaattatc gacgagcgcc tgatcaaccc cgacggctcc
     5401 ctgctgttcc gagtaaccat caacggagtg accggctggc ggctgtgcga acgcattctg
     5461 gcgtaagcgg ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     5521 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     5581 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     5641 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     5701 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc atatgctggc
     5761 tgccatgaac aaaggtggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     5821 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     5881 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     5941 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg aagatccctc
     6001 gacctgcagg ctccagccta cacaatcgct caagacgtgt aatgctttta ttatatatta
     6061 gtcacgatat ctataacaag aaaatatata tataataagt tatcacgtaa gtagaacatg
     6121 aaataacaat ataattatcg tatgagttaa atcttaaaag tcacgtaaaa gataatcatg
     6181 cgtcattttg actcacgcgg tcgttatagt tcaaaatcag tgacacttac cgcattgaca
     6241 agcacgcctc acgggagctc caagcggcga ctgagatgtc ctaaatgcac agcgacggat
     6301 tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga
     6361 ctatctttct agggttaaaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6421 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6481 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6541 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     6601 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     6661 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     6721 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     6781 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     6841 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     6901 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6961 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     7021 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     7081 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     7141 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     7201 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     7261 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     7321 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     7381 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     7441 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7501 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7561 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     7621 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7681 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7741 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7801 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     7861 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7921 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7981 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     8041 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     8101 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     8161 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     8221 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     8281 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     8341 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8401 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8461 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8521 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8581 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8641 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8701 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8761 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8821 atacatattt gaatgtattt agaaaaata
//
