LOCUS       sequence-306421-        8569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac transposon vector with CAG promoter expressing Cdk6 (R31C 
            mutant) and EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8569)
  AUTHORS   Ballabio C, Anderle M, Gianesello M, Lago C, Miele E, Cardano M, 
            Aiello G, Piazza S, Caron D, Gianno F, Ciolfi A, Pedace L, 
            Mastronuzzi A, Tartaglia M, Locatelli F, Ferretti E, Giangaspero F, 
            Tiberi L
  TITLE     Modeling medulloblastoma in vivo and with human cerebellar 
            organoids.
  JOURNAL   Nat Commun. 2020 Jan 29;11(1):583. doi: 10.1038/s41467-019-13989-3.
  PUBMED    31996670
REFERENCE   2  (bases 1 to 8569)
  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: "Nat 
            Commun."; date: "2020-01-29"; volume: "11(1):583. doi"; pages: " 
            10.1038/s41467-019-13989-3"
COMMENT     Sequence Label: pPB CAG Cdk6 R31C-IRES-EGFP
FEATURES             Location/Qualifiers
     source          1..8569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   41..103
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    447..480
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        508..887
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        889..1166
                     /label=chicken beta-actin promoter
     intron          1167..2183
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2107..2129
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2191..2210
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(3266..3840,3841..3843,3844..3852)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3433..3450)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3660..3679
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3853..3855,3856..3858,3859..4572)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3853..3855,3856..3858,3859..4569)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3898..3919)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4159..4178)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4506..4527
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4664..4683)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4729..4784
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4783..4802)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     protein_bind    5131..5164
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   5531..5565
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(5714..5732)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5753..5769)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5753..5769)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5766..5788)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5777..5793
                     /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(5801..5807,5808..5825,5826..5831))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5846..5867
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5984..6001)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6155..6743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6235..6254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6914..7705,7706..7774))
                     /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     7537..7556
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7775..7879)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7905..8360)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8042..8063)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8254..8273
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8487..8509
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8501..8518
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8502..8518
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8525..8544
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8525..8543
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agcaggctga ataataaaaa aattagaaac tattatttaa ccctagaaag ataatcatat
       61 tgtgacgtac gttaaagata atcatgcgta aaattgacgc atgtgtttta tcggtctgta
      121 tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt acacttacat
      181 actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa aaaaaaacaa
      241 aaactcaaaa tttcttctat aaagtaacaa aacttttaaa cattctctct tttacaaaaa
      301 taaacttatt ttgtacttta aaaacagtca tgttgtatta taaaataagt aattagctta
      361 acctatacat aatagaaaca aattatactt attagtcagt cagaaacaac tttggcacat
      421 atcaatatta tgctctcgtt aatcgcataa cttcgtataa tgtatgctat acgaagttat
      481 aattcagatc tttcggcgcg ccgggtcgac attgattatt gactagttat taatagtaat
      541 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      601 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      661 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      721 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      781 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      841 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc gaggtgagcc
      901 ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt ttgtatttat
      961 ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg cgcgcgccag
     1021 gcggggcggg gcggggcgag gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca
     1081 atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg cggcggccct
     1141 ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc
     1201 gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg
     1261 tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc
     1321 ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg
     1381 gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct
     1441 ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca
     1501 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg
     1561 ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt
     1621 cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt
     1681 cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
     1741 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg
     1801 ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc agccattgcc
     1861 ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc tgtgcggagc
     1921 cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg
     1981 ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc
     2041 tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca
     2101 gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc
     2161 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc
     2221 attttggcaa agaattgctc gagatggaga aggacagcct gagtcgcgcc gatcagcagt
     2281 atgagtgcgt ggcggagatc ggcgaaggcg cctatgggaa ggtgttcaag gcctgcgacc
     2341 tgaagaacgg cggccgcttc gtggctctga agcgcgtgcg agtgcagacc agtgaggagg
     2401 gcatgccgct ctccaccatc cgcgaggtgg cggtgctgag gcacctggag accttcgagc
     2461 accccaacgt ggtcaggttg tttgatgtgt gcacagtgtc acggacggac agagaaacca
     2521 agcttacact agtgtttgag catgttgatc aagacttgac cacttacttg gataaagttc
     2581 cagagcccgg cgtacccaca gaaaccataa aggatatgat gtttcagctt ctccgaggtc
     2641 tggactttct tcattctcac agagtagtgc atcgtgatct gaaaccgcag aacattctgg
     2701 tgaccagcag tggacagata aagctggctg actttggcct tgcccgcatc tatagttttc
     2761 agatggccct tacctcggtg gtcgtcacgc tgtggtaccg agccccagaa gtcctgctcc
     2821 agtccagcta tgccacccct gtggacctct ggagtgtcgg ttgcatcttt gcagaaatgt
     2881 ttcgcagaaa gcctcttttt cgtggaagtt cagacgtgga tcaactagga aaaatcttgg
     2941 acatcattgg actcccagga gaggaagact ggcctaggga cgtggccctt ccccggcagg
     3001 cttttcattc caaatctgct caacccatcg agaagtttgt gacagatatt gacgaactag
     3061 gcaaagacct acttctgaaa tgcctgacgt ttaatccagc taaaaggata tccgcctacg
     3121 gcgccctgaa tcacccgtac ttccaagatc tggagagata caaggacaac ctgaactctc
     3181 acctgccatc caaccagagc acctcggagc tgaacacagc ctgatgagaa ttctgcagtc
     3241 gacggtaccg cgggcccggg atccgcccct ctccctcccc cccccctaac gttactggcc
     3301 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     3361 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     3421 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     3481 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     3541 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     3601 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     3661 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     3721 tgggatctga tctggggcct cggtacacat gctttacatg tgtttagtcg aggttaaaaa
     3781 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     3841 atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3901 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3961 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     4021 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     4081 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     4141 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     4201 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     4261 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     4321 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     4381 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     4441 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     4501 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     4561 ctgtacaagt aaagcggccg caattcactc ctcaggtgca ggctgcctat cagaaggtgg
     4621 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     4681 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     4741 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     4801 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     4861 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     4921 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     4981 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     5041 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     5101 tatggagatc cctcgacctg caccgtcgag ataacttcgt ataatgtatg ctatacgaag
     5161 ttatgctagc caacaagctc gtcatcgctt tgcagaagag cagagaggat atgctcatcg
     5221 tctaaagaac tacccatttt attatatatt agtcacgata tctataacaa gaaaatatat
     5281 atataataag ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta
     5341 aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag
     5401 ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg
     5461 actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat
     5521 ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaaa aaagatttgc
     5581 gctttactcg acctaaactt taaacacgtc atagaatctt cgtttgacaa aaaccacatt
     5641 gtggccaagc tgtgtgacgc gacgcgcgct aaagaatggc aaaccaagtc gcgcgaggta
     5701 ccagcttttg ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc
     5761 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5821 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5881 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5941 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     6001 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     6061 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     6121 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6181 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6241 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6301 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6361 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6421 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6481 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6541 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6601 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6661 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6721 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6781 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6841 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6901 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6961 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     7021 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     7081 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     7141 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     7201 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     7261 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     7321 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7381 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7441 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7501 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7561 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7621 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7681 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7741 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7801 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     7921 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     7981 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     8041 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     8101 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     8161 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     8221 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     8281 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     8341 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     8401 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     8461 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     8521 attgtaatac gactcactat agggcgaatt ggagctcggt attcacgac
//
