LOCUS       sequence-492505-        8231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses photoactivatable H2B-Cherry under EF1 promoter and can be 
            randomly integrated by PiggyBac transposase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8231)
  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 8231)
  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-EF1-alpha-H2B-PA-mCherry-PGKneo
FEATURES             Location/Qualifiers
     source          1..8231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     372..391
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             453..1256
                     /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(516..535)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1126..1145
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1263..1311
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    1330..1363
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   1610..1672
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(1802..1820)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1841..1857)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1841..1857)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1854..1876)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1865..1881
                     /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(1889..1895,1896..1913,1914..1919))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1934..1955
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2072..2089)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2243..2831)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2323..2342)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3002..3793,3794..3862))
                     /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     3625..3644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3863..3967)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3993..4448)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4130..4151)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4342..4361
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4575..4597
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4589..4606
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4590..4606
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4613..4631
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   4808..4842
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        5139..6317
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5370..6308
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6265..6285
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        6334..6352
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6436..6813
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      6826..6835
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6832..7542
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein"
                     /label=PAmCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6977..6995)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7245..7264
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7609..7650
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(7688..7705)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7694..7918
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7947..7963)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7947..7963)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     8013..8031
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    8076..8109
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(8171..8231,1..439)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8206..8229)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
       61 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
      121 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
      181 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
      241 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
      301 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
      361 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
      421 ctcatctccg ggcctttcga cctgcagcca atatgggatc ggccattgaa caagatggat
      481 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
      541 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
      601 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
      661 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
      721 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
      781 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
      841 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
      901 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
      961 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     1021 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     1081 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     1141 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     1201 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagggg
     1261 atcggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cggatccgga
     1321 acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtcc ctcgaagagg
     1381 ttcactagta ctggccattg cggccttaat taaggtaccg gcgcgccgtt taaacgggcc
     1441 ctttgttact ttatagaaga aattttgagt ttttgttttt ttttaataaa taaataaaca
     1501 taaataaatt gtttgttgaa tttattatta gtatgtaagt gtaaatataa taaaacttaa
     1561 tatctattca aattaataaa taaacctcga tatacagacc gataaaacac atgcgtcaat
     1621 tttacgcatg attatcttta acgtacgtca caatatgatt atctttctag ggttaaataa
     1681 tagtttctaa tttttttatt attcagcctg ctgtcgtgaa taccgagctc gctaccttag
     1741 gaccgttata gttaattaag gcgcgccatt accctgttat ccctagctcc agcttttgtt
     1801 ccctttagtg agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1861 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1921 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1981 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     2041 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     2101 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     2161 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     2221 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     2281 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     2341 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2401 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2461 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2521 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2581 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2641 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2701 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2761 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2821 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2881 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2941 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3961 ttccgcgcac atttccccga aaagtgccac ctaaattgta agcgttaata ttttgttaaa
     4021 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     4081 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     4141 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     4201 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     4261 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     4321 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4381 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4441 gggcgcgtcc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     4501 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     4561 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga
     4621 ctcactatag ggcgaattgg gtacgcgtgc gatcgcgcta ccttaagaga ggtacctcgc
     4681 gcgacttggt ttgccattct ttagcgcgcg tcgcgtcaca cagcttggcc acaatgtggt
     4741 ttttgtcaaa cgaagattct atgacgtgtt taaagtttag gtcgagtaaa gcgcaaatct
     4801 tttttaaccc tagaaagata gtctgcgtaa aattgacgca tgcattcttg aaatattgct
     4861 ctctctttct aaatagcgcg aatccgtcgc tgtgcattta ggacatctca gtcgccgctt
     4921 ggagctcccg tgaggcgtgc ttgtcaatgc ggtaagtgtc actgattttg aactataacg
     4981 accgcgtgag tcaaaatgac gcatgattat cttttacgtg acttttaaga tttaactcat
     5041 acgataatta tattgttatt tcatgttcta cttacgtgat aacttattat atatatattt
     5101 tcttgttata gatatcggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     5161 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaaccg
     5221 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     5281 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     5341 ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc cgcgggcctg
     5401 gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg ctgcagtacg
     5461 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt
     5521 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg
     5581 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt
     5641 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg
     5701 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg
     5761 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac
     5821 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg
     5881 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc
     5941 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg
     6001 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg
     6061 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta
     6121 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg
     6181 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt
     6241 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc
     6301 catttcaggt gtcgtgagga attagcttgg tactaatacg actcactata gggagaccca
     6361 agctggctag gtaagcttgg taccgagctc ggatccacta gtccagtgtg gtggaattct
     6421 gcagatctag ctagcatgcc agagccagcg aagtctgctc ccgccccgaa aaagggctcc
     6481 aagaaggcgg tgactaaggc gcagaagaaa ggcggcaaga agcgcaagcg cagccgcaag
     6541 gagagctatt ccatctatgt gtacaaggtt ctgaagcagg tccaccctga caccggcatt
     6601 tcgtccaagg ccatgggcat catgaattcg tttgtgaacg acattttcga gcgcatcgca
     6661 ggtgaggctt cccgcctggc gcattacaac aagcgctcga ccatcacctc cagggagatc
     6721 cagacggccg tgcgcctgct gctgcctggg gagttggcca agcacgccgt gtccgagggt
     6781 actaaggcca tcaccaagta caccagcgct aaggatccac cggtcgccac catggtgagc
     6841 aagggcgagg aggataacat ggccatcatt aaggagttca tgcgcttcaa ggtgcacatg
     6901 gaggggtccg tgaacggcca cgtgttcgag atcgagggcg agggcgaggg ccgcccctac
     6961 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcacctgg
     7021 gacatcctgt cccctcaatt catgtacggc tccaatgcct acgtgaagca ccccgccgac
     7081 atccccgact actttaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaaa
     7141 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggtgagttc
     7201 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     7261 aagaccatgg gctgggaggc cctctccgag cggatgtacc ccgaggacgg cgccctgaag
     7321 ggcgaggtca agccgagagt gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     7381 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caaccgcaag
     7441 ttggacatca cctcacacaa cgaggactac accatcgtgg aacagtacga acgtgccgag
     7501 ggccgccact ccaccggcgg catggacgag ctgtacaagt aagcggccgc ttttttcctt
     7561 atccagcaca gtggcggccg ctcgagtcta gagggcccgc ggttcgaagg taagcctatc
     7621 cctaaccctc tcctcggtct cgattctacg cgtaccggtt agtaatgagt ttaaacccgc
     7681 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     7741 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     7801 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     7861 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     7921 tgatatcgaa ttcctgcagc ccgggggatc cactagttct agagcggccg cacgtctaag
     7981 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     8041 ttcaagaatt ccgatcatat tcaataaccc ttaatataac ttcgtataat gtatgctata
     8101 cgaagttatt aggtctgaag aggagtttac gtccagccaa gcttaggggc ccctgcaggt
     8161 caattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca
     8221 gccccgctgg g
//
