LOCUS       sequence-493810-        9061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of NLS-MARIO, a genetically-encoded fluorescent
            Mg2+ sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9061)
  AUTHORS   Maeshima K, Matsuda T, Shindo Y, Imamura H, Tamura S, Imai R, 
            Kawakami S, Nagashima R, Soga T, Noji H, Oka K, Nagai T
  TITLE     A Transient Rise in Free Mg(2+) Ions Released from ATP-Mg Hydrolysis
            Contributes to Mitotic Chromosome Condensation.
  JOURNAL   Curr Biol. 2018 Feb 5;28(3):444-451.e6. doi: 
            10.1016/j.cub.2017.12.035. Epub 2018 Jan 18.
  PUBMED    29358072
REFERENCE   2  (bases 1 to 9061)
  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: 
            "10.1016/j.cub.2017.12.035"; journalName: "Curr Biol"; date: 
            "2018-02-5- 5"; volume: "28"; issue: "3"; pages: "444-451.e6"
COMMENT     Sequence Label: pPB-PGKneo-EF1a-NLS-MARIO-SV40 pA
FEATURES             Location/Qualifiers
     source          1..9061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     308..331
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    complement(428..461)
                     /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(506..524)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     574..590
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     574..590
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    598..719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(609..628)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     663..682
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(856..1590)
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTAVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     CDS             complement(join(2359..2871,2872..2874,2875..2877))
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     CDS             complement(2899..2919)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(3015..3033)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3050..4228)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3059..3997
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(3082..3102)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     repeat_region   4525..4559
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(4736..4754)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4761..4778)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4761..4777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4770..4792)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4919..5374
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5006..5025)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5216..5237
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5400..5504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5505..5573,5574..6365)
                     /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(5723..5742)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6536..7124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7025..7044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7278..7295
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7412..7433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7448..7453,7454..7471,7472..7478)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7486..7502
                     /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     7491..7513
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7510..7526
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7510..7526
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7547..7565
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   7695..7757
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    complement(8004..8037)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    8056..8104
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(8111..8914)
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8222..8241)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8832..8851
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(8928..9061,1..366))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8976..8995)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 acccacccct tcccagcctc tgagcccaga aagcgaagga gcaaagctgc tattggccgc
       61 tgccccaaag gcctacccgc ttccattgct cagcggtgct gtccatctgc acgagactag
      121 tgagacgtgc tacttccatt tgtcacgtcc tgcacgacgc gagctgcggg gcggggggga
      181 acttcctgac taggggagga gtagaaggtg gcgcgaaggg gccaccaaag aacggagccg
      241 gttggcgcct accggtggat gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc
      301 aagtgcccag cggggctgct aaagcgcatg ctccagactg ccttgggaaa agcgcctccc
      361 ctacccggta gaattgacct gcaggggccc ctaagcttgg ctggacgtaa actcctcttc
      421 agacctaata acttcgtata gcatacatta tacgaagtta tattaagggt tattgaatat
      481 gatcggaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
      541 atgataataa tggtttctta gacgtgcggc cgctctagaa ctagtggatc ccccgggtaa
      601 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      661 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
      721 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
      781 aaagcaagta aaacctctac aaatgtggta tggctgatta tgatcagtta tctagatccg
      841 gtggatcaat tcttactcga tgttgtggcg gatcttgaag ttggccttga tgccgttctt
      901 ctgcttgtcg gcggtgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
      961 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     1021 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
     1081 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     1141 gtggtcgggg tagcgggcga agcactgcag gccgtagccc agggtggtca cgagggtggg
     1201 ccagggcacg ggcagcttgc cggtggtgca gatcagcttc agggtcagct tgccgtaggt
     1261 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
     1321 ctcgaccagg atgggcacca ccgcggtgaa cagctcctcg cccttgctca ccatgccacc
     1381 gctgccaccc ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
     1441 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actggtagct
     1501 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
     1561 gtggtcggcg agctgcacgc cgccgtccat gagctcagcc gcctgcatca ggaagttcac
     1621 cttctggaac agggattcgt tatgcggcag cagggattcg atatctcgca ggatttcacg
     1681 cgcctgctcc agttgcccac ccggtaaacg cgctttacgc accaggaagt tgagcgcgcg
     1741 ctgggtatcc atcagacaca gggcaacggc ccagccgtca tcttccagtt ccgccagagt
     1801 ggagagcgcc tcgtcgtact catcgccctg atgcccttcc ataatcaccc ggctcaactg
     1861 ctccaggtcg ctatagatat tttcaatttc atctgccaac tgttcgattt tggtttcgaa
     1921 cagatccagc agcaactcgt aggcgttacc gtctaccatc gactggctac gggcacgcat
     1981 acgatacaga cgaaaagcgg gcagttcacg ctcacgcaga gtaaacagac gaccatcacg
     2041 gatggtaaat gccacagtgg agttaccggc gtgatcttcc gcatcttcaa agaagaagaa
     2101 ggagtgatca tgcaggccgt cgtcgtcttc aaagaaacgt gccgatgctt cgatgtcttc
     2161 cagttccggg cgggttgcca ggctcagcat gcgggcggcg gtcacgaact ccagcaggac
     2221 catgtgatcg cgcttctcgt tggggtcttt gctcagggcg gactgggtgc tcaggtagtg
     2281 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
     2341 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagtgggcct tgatgccgtt
     2401 cttctgcttg tcggcggtga tatagacgtt gtggctgatg tagttgtact ccagcttgtg
     2461 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     2521 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
     2581 gatggtacgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     2641 catgtggtcg gggtagcggc tgaagcactg cacgccccag gtcagggtgg tcacgagggt
     2701 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     2761 ggtggcatcg ccctcgccct cgccggacac gctgaacctg tggccgttta cgtcgccgtc
     2821 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggg
     2881 atcgatcctg gggtcttcta cctttctctt cttttttgga ggccccccca tgctagctag
     2941 atctgcagaa ttccaccaca ctggactagt ggatccgagc tcggtaccaa gcttacctag
     3001 ccagcttggg tctccctata gtgagtcgta ttagtaccaa gctaattcct cacgacacct
     3061 gaaatggaag aaaaaaactt tgaaccactg tctgaggctt gagaatgaac caagatccaa
     3121 actcaaaaag ggcaaattcc aaggagaatt acatcaagtg ccaagctggc ctaacttcag
     3181 tctccaccca ctcagtgtgg ggaaactcca tcgcataaaa cccctccccc caacctaaag
     3241 acgacgtact ccaaaagctc gagaactaat cgaggtgcct ggacggcgcc cggtactccg
     3301 tggagtcaca tgaagcgacg gctgaggacg gaaaggccct tttcctttgt gtgggtgact
     3361 cacccgcccg ctctcccgag cgccgcgtcc tccattttga gctccctgca gcagggccgg
     3421 gaagcggcca tctttccgct cacgcaactg gtgccgaccg ggccagcctt gccgcccagg
     3481 gcggggcgat acacggcggc gcgaggccag gcaccagagc aggccggcca gcttgagact
     3541 acccccgtcc gattctcggt ggccgcgctc gcaggccccg cctcgccgaa catgtgcgct
     3601 gggacgcacg ggccccgtcg ccgcccgcgg ccccaaaaac cgaaatacca gtgtgcagat
     3661 cttggcccgc atttacaaga ctatcttgcc agaaaaaaag cgtcgcagca ggtcatcaaa
     3721 aattttaaat ggctagagac ttatcgaaag cagcgagaca ggcgcgaagg tgccaccaga
     3781 ttcgcacgcg gcggccccag cgcccaggcc aggcctcaac tcaagcacga ggcgaagggg
     3841 ctccttaagc gcaaggcctc gaactctccc acccacttcc aacccgaagc tcgggatcaa
     3901 gaatcacgta ctgcagccag gtggaagtaa ttcaaggcac gcaagggcca taacccgtaa
     3961 agaggccagg cccgcgggaa ccacacacgg cacttacctg tgttctggcg gcaaacccgt
     4021 tgcgaaaaag aacgttcacg gcgactactg cacttatata cggttctccc ccaccctcgg
     4081 gaaaaaggcg gagccagtac acgacatcac tttcccagtt taccccgcgc caccttctct
     4141 aggcaccggt tcaattgccg acccctcccc ccaacttctc ggggactgtg ggcgatgtgc
     4201 gctctgccca ctgacgggca ccggagccaa ttcccactcc tttcaagacc gatatctata
     4261 acaagaaaat atatatataa taagttatca cgtaagtaga acatgaaata acaatataat
     4321 tatcgtatga gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca
     4381 cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg
     4441 agctccaagc ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa
     4501 agagagagca atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt
     4561 taaaaaagat ttgcgcttta ctcgacctaa actttaaaca cgtcatagaa tcttcgtttg
     4621 acaaaaacca cattgtggcc aagctgtgtg acgcgacgcg cgctaaagaa tggcaaacca
     4681 agtcgcgcga ggtacctctc ttaaggtagc gcgatcgcac gcgtacccaa ttcgccctat
     4741 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     4801 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4861 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     4921 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4981 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     5041 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     5101 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5161 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5221 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5281 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5341 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     5401 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5881 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5941 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6661 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6901 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7021 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7081 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7141 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7201 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7261 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     7321 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     7381 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     7441 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     7501 aatttcacac aggaaacagc tatgaccatg attacgccaa gctcgaaatt aaccctcact
     7561 aaagggaaca aaagctggag ctagggataa cagggtaatg gcgcgcctta attaactata
     7621 acggtcctaa ggtagcgagc tcggtattca cgacagcagg ctgaataata aaaaaattag
     7681 aaactattat ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg
     7741 cgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa
     7801 tagatattaa gttttattat atttacactt acatactaat aataaattca acaaacaatt
     7861 tatttatgtt tatttattta ttaaaaaaaa acaaaaactc aaaatttctt ctataaagta
     7921 acaaagggcc cgtttaaacg gcgcgccggt accttaatta aggccgcaat ggccagtact
     7981 agtgaacctc ttcgagggac ctaataactt cgtatagcat acattatacg aagttatatt
     8041 aagggttccg gatccgaaca aacgacccaa cacccgtgcg ttttattctg tctttttatt
     8101 gccgatcccc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     8161 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     8221 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     8281 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     8341 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     8401 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     8461 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     8521 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     8581 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     8641 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     8701 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     8761 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     8821 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     8881 cgtgcaatcc atcttgttca atggccgatc ccatattggc tgcaggtcga aaggcccgga
     8941 gatgaggaag aggagaacag cgcggcagac gtgcgctttt gaagcgtgca gaatgccggg
     9001 cctccggagg accttcgggc gcccgccccg cccctgagcc cgcccctgag cccgcccccg
     9061 g
//
