LOCUS       sequence-408509-       11358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dox-inducible overexpression of HA-tagged KRASG12D and mCherry in 
            human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11358)
  AUTHORS   Breunig M, Merkle J, Wagner M, Melzer MK, Barth TFE, Engleitner T, 
            Krumm J, Wiedenmann S, Cohrs CM, Perkhofer L, Jain G, Kruger J, 
            Hermann PC, Schmid M, Madacsy T, Varga A, Griger J, Azoitei N, 
            Muller M, Wessely O, Robey PG, Heller S, Dantes Z, Reichert M, Gunes
            C, Bolenz C, Kuhn F, Maleth J, Speier S, Liebau S, Sipos B, Kuster 
            B, Seufferlein T, Rad R, Meier M, Hohwieler M, Kleger A
  TITLE     Modeling plasticity and dysplasia of pancreatic ductal organoids 
            derived from human pluripotent stem cells.
  JOURNAL   Cell Stem Cell. 2021 Jun 3;28(6):1105-1124.e19. doi: 
            10.1016/j.stem.2021.03.005. Epub 2021 Apr 28.
  PUBMED    33915078
REFERENCE   2  (bases 1 to 11358)
  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.stem.2021.03.005"; journalName: "Cell Stem Cell"; date: 
            "2021-06-3- 3"; volume: "28"; issue: "6"; pages: "1105-1124.e19"
COMMENT     Sequence Label: PB-TAC-ERP2-(N-HA)KRAS_G12D
FEATURES             Location/Qualifiers
     source          1..11358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     597..614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     621..640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   706..768
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1140..1158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1177..1195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1214..1232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1251..1269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1288..1306
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1325..1343
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1362..1380
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1399..1417
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1439..1463
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1543..1567
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1594..1620
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1600..1626
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             join(1648..1680,1681..1683,1684..2214)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             1648..2214
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(2240..2264)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(2306..2880,2881..2883,2884..2892)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2893..3603
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3038..3056)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3306..3325
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3615..3634)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3680..3735
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3734..3753)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        4178..4557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4672..5977
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    5014..5945
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(5994..6611,6612..6617,6618..6653,6654..6656,6657..6692,
                     6693..6695,6696..6731,6732..6740)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    join(6767..7341,7342..7344,7345..7353)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             7354..7953
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7354..7373)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7850..7870
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8039..8611
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   8985..9019
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(9168..9186)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9207..9223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9207..9223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9220..9242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9231..9247
                     /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(9255..9261,9262..9279,9280..9285))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9300..9321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9438..9455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9609..10197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9689..9708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10368..11159,11160..11228))
                     /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     10991..11010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11229..11333)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcggtattc
      661 acgacagcag gctgaataat aaaaaaatta gaaactatta tttaacccta gaaagataat
      721 catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt
      781 ctgtatatcg aggtttattt attaatttga atagatatta agttttatta tatttacact
      841 tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa
      901 aacaaaaact caaaatttct tctataaagt aacaaaactt ttaaacattc tctcttttac
      961 aaaaataaac ttattttgta ctttaaaaac agtcatgttg tattataaaa taagtaatta
     1021 gcttaaccta tacataatag aaacaaatta tacttattag tcagtcagaa acaactttgg
     1081 cacatatcaa tattatgctc tcgttaatta actcgaggga tccaagctgg ccgcacgtct
     1141 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     1201 cgtctagaac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     1261 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     1321 gagctcccta tcagtgatag agaaggtacg tctagaacgt ctccctatca gtgatagaga
     1381 agtcgacacg tctcgagctc cctatcagtg atagagaagg taccccctat ataagcagag
     1441 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1501 gaagacaccg ggaccgatcc agcctccgcg gtggcggcca tcacaagttt gtacaaaaaa
     1561 gcaggcttca ctagtgtctt tcataagctt atgtatccat atgatgtgcc cgactatgcc
     1621 tcacttggag gaccctctgg atccaccatg actgaatata aacttgtggt agttggagct
     1681 gacggcgtag gcaagagtgc cttgacgata cagctaattc agaatcattt tgtggacgaa
     1741 tatgatccaa caatagagga ttcctacagg aagcaagtag taattgatgg agaaacctgt
     1801 ctcttggata ttctcgacac agcaggtcaa gaggagtaca gtgcaatgag ggaccagtac
     1861 atgaggactg gggagggctt tctttgtgta tttgccataa ataatactaa atcatttgaa
     1921 gatattcacc attatagaga acaaattaaa agagttaagg actctgaaga tgtacctatg
     1981 gtcctagtag gaaataaatg tgatttgcct tccagaacag tagacacaaa acaggctcag
     2041 gacttagcaa gaagttatgg aattcctttt attgaaacat cagcaaagac aagacagggt
     2101 gttgatgatg ccttctatac attagttcga gaaattcgaa aacataaaga aaagatgagc
     2161 aaagatggta aaaagaagaa aaagaagtca aagacaaagt gtgtaattat gtaagtcgac
     2221 cctgtggaat gtgtgtcaca cccagctttc ttgtacaaag tggtgatcta gctaaaacgc
     2281 ggcctcgaat cgaattccgc ccccccccct ctccctcccc cccccctaac gttactggcc
     2341 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     2401 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     2461 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     2521 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     2581 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     2641 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     2701 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     2761 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     2821 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     2881 atggccacaa ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     2941 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     3001 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     3061 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     3121 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     3181 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     3241 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     3301 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     3361 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     3421 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     3481 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     3541 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     3601 taaagcggcc gcgatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     3661 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3721 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     3781 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     3841 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     3901 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     3961 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     4021 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcaggaattc
     4081 gattacttaa gcctaggatc cagatctgta cattcgaatc gatgaattcg ttaaccatgg
     4141 aagcttctgc agcggccgca ctagcggtgg cggcctcgac attgattatt gactagttat
     4201 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     4261 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     4321 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     4381 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     4441 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     4501 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatgatg
     4561 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     4621 agtctccacc ccattgacgt caatgggagt ttgttttgac tagtggagcc gagagtaatt
     4681 catacaaaag gagggatcgc cttcgcaagg ggagagccca gggaccgtcc ctaaattctc
     4741 acagacccaa atccctgtag ccgccccacg acagcgcgag gagcatgcgc ccagggctga
     4801 gcgcgggtag atcagagcac acaagctcac agtccccggc ggtgggggga ggggcgcgct
     4861 gagcgggggc cagggagctg gcgcggggca aactgggaaa gtggtgtcgt gtgctggctc
     4921 cgccctcttc ccgagggtgg gggagaacgg tatataagtg cggtagtcgc cttggacgtt
     4981 ctttttcgca acgggtttgc cgtcagaacg caggtgagtg gcgggtgtgg cttccgcggg
     5041 ccccggagct ggagccctgc tctgagcggg ccgggctgat atgcgagtgt cgtccgcagg
     5101 gtttagctgt gagcattccc acttcgagtg gcgggcggtg cgggggtgag agtgcgaggc
     5161 ctagcggcaa ccccgtagcc tcgcctcgtg tccggcttga ggcctagcgt ggtgtccgcc
     5221 gccgcgtgcc actccggccg cactatgcgt tttttgtcct tgctgccctc gattgccttc
     5281 cagcagcatg ggctaacaaa gggagggtgt ggggctcact cttaaggagc ccatgaagct
     5341 tacgttggat aggaatggaa gggcaggagg ggcgactggg gcccgcccgc cttcggagca
     5401 catgtccgac gccacctgga tggggcgagg cctgtggctt tccgaagcaa tcgggcgtga
     5461 gtttagccta cctgggccat gtggccctag cactgggcac ggtctggcct ggcggtgccg
     5521 cgttcccttg cctcccaaca agggtgaggc cgtcccgccc ggcaccagtt gcttgcgcgg
     5581 aaagatggcc gctcccgggg ccctgttgca aggagctcaa aatggaggac gcggcagccc
     5641 ggtggagcgg gcgggtgagt cacccacaca aaggaagagg gccttgcccc tcgccggccg
     5701 ctgcttcctg tgaccccgtg gtctatcggc cgcatagtca cctcgggctt ctcttgagca
     5761 ccgctcgtcg cggcgggggg aggggatcta atggcgttgg agtttgttca catttggtgg
     5821 gtggagacta gtcaggccag cctggcgctg gaagtcattc ttggaatttg cccctttgag
     5881 tttggagcga ggctaattct caagcctctt agcggttcaa aggtattttc taaacccgtt
     5941 tccaggtgtt gtgaaagcca ccgctaattc aaagcaatcc ggagtaattc accatgtcta
     6001 gactggacaa gagcaaagtc ataaacggcg ctctggaatt actcaatgga gtcggtatcg
     6061 aaggcctgac gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact
     6121 ggcacgtgaa gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc
     6181 atcataccca cttctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg
     6241 ccaagtcatt ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca
     6301 cccgcccaac agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc
     6361 aaggcttctc cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg
     6421 gctgcgtatt ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca
     6481 ccgattctat gcccccactt ctgagacaag caattgagct gttcgaccgg cagggagccg
     6541 aacctgcctt ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt
     6601 gcgaaagcgg cgggccggcc gacgcccttg acgattttga cttagacatg ctcccagccg
     6661 atgcccttga cgactttgac cttgatatgc tgcctgctga cgctcttgac gattttgacc
     6721 ttgacatgct ccccgggtaa ctaagtaagg atcattccgc cccccccccc tctccctccc
     6781 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     6841 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     6901 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     6961 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     7021 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     7081 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     7141 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     7201 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     7261 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     7321 tttgaaaaac acgatgataa tatggccaca accatgaccg agtacaagcc cacggtgcgc
     7381 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     7441 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     7501 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     7561 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     7621 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     7681 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     7741 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     7801 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     7861 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     7921 acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg
     7981 accgaaagga gcgcacgacc ccatgcatcg aaatgaccga ccaagcgaat tcgctaggat
     8041 tatccctaat acctgccacc ccactcttaa tcagtggtgg aagaacggtc tcagaactgt
     8101 ttgtttcaat tggccattta agtttagtag taaaagactg gttaatgata acaatgcatc
     8161 gtaaaacctt cagaaggaaa ggagaatgtt ttgtggacca ctttggtttt cttttttgcg
     8221 tgtggcagtt ttaagttatt agtttttaaa atcagtactt tttaatggaa acaacttgac
     8281 caaaaatttg tcacagaatt ttgagaccca ttaaaaaagt taaatgagaa acctgtgtgt
     8341 tcctttggtc aacaccgaga catttaggtg aaagacatct aattctggtt ttacgaatct
     8401 ggaaacttct tgaaaatgta attcttgagt taacacttct gggtggagaa tagggttgtt
     8461 ttccccccac ataattggaa ggggaaggaa tatcatttaa agctatggga gggttgcttt
     8521 gattacaaca ctggagagaa atgcagcatg ttgctgattg cctgtcacta aaacaggcca
     8581 aaaactgagt ccttgggttg catagaaagc tgccaaacgc tagccaacaa gctcgtcatc
     8641 gctttgcaga agagcagaga ggatatgctc atcgtctaaa gaactaccca ttttattata
     8701 tattagtcac gatatctata acaagaaaat atatatataa taagttatca cgtaagtaga
     8761 acatgaaata acaatataat tatcgtatga gttaaatctt aaaagtcacg taaaagataa
     8821 tcatgcgtca ttttgactca cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat
     8881 tgacaagcac gcctcacggg agctccaagc ggcgactgag atgtcctaaa tgcacagcga
     8941 cggattcgcg ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac
     9001 gcagactatc tttctagggt taaaaaagat ttgcgcttta ctcgacctaa actttaaaca
     9061 cgtcatagaa tcttcgtttg acaaaaacca cattgtggcc aagctgtgtg acgcgacgcg
     9121 cgctaaagaa tggcaaacca agtcgcgcga ggtaccagct tttgttccct ttagtgaggg
     9181 ttaatttcga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     9241 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     9301 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     9361 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     9421 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     9481 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     9541 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     9601 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     9661 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     9721 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     9781 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     9841 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     9901 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     9961 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
    10021 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
    10081 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
    10141 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
    10201 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
    10261 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
    10321 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
    10381 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
    10441 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
    10501 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
    10561 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
    10621 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
    10681 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
    10741 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
    10801 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
    10861 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
    10921 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
    10981 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
    11041 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
    11101 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
    11161 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
    11221 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
    11281 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
    11341 ccccgaaaag tgccacct
//
