LOCUS       sequence-347424-       13143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac vector for DOX-inducible human NICD-IRES-EGFP expression in
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13143)
  AUTHORS   Chu LF, Mamott D, Ni Z, Bacher R, Liu C, Swanson S, Kendziorski C, 
            Stewart R, Thomson JA
  TITLE     An In Vitro Human Segmentation Clock Model Derived from Embryonic 
            Stem Cells.
  JOURNAL   Cell Rep. 2019 Aug 27;28(9):2247-2255.e5. doi: 
            10.1016/j.celrep.2019.07.090.
  PUBMED    31461642
REFERENCE   2  (bases 1 to 13143)
  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.celrep.2019.07"; journalName: "Cell Rep"; date: 
            "2019-08-27- 27"; volume: "28"; issue: "9"; pages: "2247-2255.e5"
COMMENT     Sequence Label: pB-TAG-NICD
FEATURES             Location/Qualifiers
     source          1..13143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   86..148
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    520..538
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    557..575
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    594..612
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    631..649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    668..686
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    705..723
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    742..760
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    779..797
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     819..843
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    923..947
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3396..3420)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    join(3462..4036,4037..4039,4040..4048)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             join(4049..4051,4052..4054,4055..4768)
                     /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(4049..4051,4052..4054,4055..4765)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4094..4115)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4355..4374)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4702..4723
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4780..4799)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4845..4900
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4899..4918)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        5343..5722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5837..7142
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    6179..7110
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(7159..7776,7777..7782,7783..7818,7819..7821,7822..7857,
                     7858..7860,7861..7896,7897..7905)
                     /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(7932..8506,8507..8509,8510..8518)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             8519..9118
                     /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(8519..8538)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9015..9035
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    9204..9776
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   10150..10184
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(10333..10351)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10372..10388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10372..10388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10385..10407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10396..10412
                     /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(10420..10426,10427..10444,10445..10450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10465..10486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10603..10620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10774..11362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10854..10873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11533..12324,12325..12393))
                     /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     12156..12175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12394..12498)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(12524..12979)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12661..12682)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12873..12892
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13106..13128
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13120..13137
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     13121..13137
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattggag ctcggtattc acgacagcag gctgaataat
       61 aaaaaaatta gaaactatta tttaacccta gaaagataat catattgtga cgtacgttaa
      121 agataatcat gcgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg aggtttattt
      181 attaatttga atagatatta agttttatta tatttacact tacatactaa taataaattc
      241 aacaaacaat ttatttatgt ttatttattt attaaaaaaa aacaaaaact caaaatttct
      301 tctataaagt aacaaaactt ttaaacattc tctcttttac aaaaataaac ttattttgta
      361 ctttaaaaac agtcatgttg tattataaaa taagtaatta gcttaaccta tacataatag
      421 aaacaaatta tacttattag tcagtcagaa acaactttgg cacatatcaa tattatgctc
      481 tcgttaatta actcgaggga tccaagctgg ccgcacgtct ccctatcagt gatagagaag
      541 tcgacacgtc tcgagctccc tatcagtgat agagaaggta cgtctagaac gtctccctat
      601 cagtgataga gaagtcgaca cgtctcgagc tccctatcag tgatagagaa ggtacgtcta
      661 gaacgtctcc ctatcagtga tagagaagtc gacacgtctc gagctcccta tcagtgatag
      721 agaaggtacg tctagaacgt ctccctatca gtgatagaga agtcgacacg tctcgagctc
      781 cctatcagtg atagagaagg taccccctat ataagcagag ctcgtttagt gaaccgtcag
      841 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      901 agcctccgcg gtggcggcca tcacaagttt gtacaaaaaa gcaggctccg cggccgcccc
      961 cttcaccatg gtgctgctgt cccgcaagcg ccggcggcag catggccagc tctggttccc
     1021 tgagggcttc aaagtgtctg aggccagcaa gaagaagcgg cgggagcccc tcggcgagga
     1081 ctccgtgggc ctcaagcccc tgaagaacgc ttcagacggt gccctcatgg acgacaacca
     1141 gaatgagtgg ggggacgagg acctggagac caagaagttc cggttcgagg agcccgtggt
     1201 tctgcctgac ctggacgacc agacagacca ccggcagtgg actcagcagc acctggatgc
     1261 cgctgacctg cgcatgtctg ccatggcccc cacaccgccc cagggtgagg ttgacgccga
     1321 ctgcatggac gtcaatgtcc gcgggcctga tggcttcacc ccgctcatga tcgcctcctg
     1381 cagcgggggc ggcctggaga cgggcaacag cgaggaagag gaggacgcgc cggccgtcat
     1441 ctccgacttc atctaccagg gcgccagcct gcacaaccag acagaccgca cgggcgagac
     1501 cgccttgcac ctggccgccc gctactcacg ctctgatgcc gccaagcgcc tgctggaggc
     1561 cagcgcagat gccaacatcc aggacaacat gggccgcacc ccgctgcatg cggctgtgtc
     1621 tgccgacgca caaggtgtct tccagatcct gatccggaac cgagccacag acctggatgc
     1681 ccgcatgcat gatggcacga cgccactgat cctggctgcc cgcctggccg tggagggcat
     1741 gctggaggac ctcatcaact cacacgccga cgtcaacgcc gtagatgacc tgggcaagtc
     1801 cgccctgcac tgggccgccg ccgtgaacaa tgtggatgcc gcagttgtgc tcctgaagaa
     1861 cggggctaac aaagatatgc agaacaacag ggaggagaca cccctgtttc tggccgcccg
     1921 ggagggcagc tacgagaccg ccaaggtgct gctggaccac tttgccaacc gggacatcac
     1981 ggatcatatg gaccgcctgc cgcgcgacat cgcacaggag cgcatgcatc acgacatcgt
     2041 gaggctgctg gacgagtaca acctggtgcg cagcccgcag ctgcacggag ccccgctggg
     2101 gggcacgccc accctgtcgc ccccgctctg ctcgcccaac ggctacctgg gcagcctcaa
     2161 gcccggcgtg cagggcaaga aggtccgcaa gcccagcagc aaaggcctgg cctgtggaag
     2221 caaggaggcc aaggacctca aggcacggag gaagaagtcc caggacggca agggctgcct
     2281 gctggacagc tccggcatgc tctcgcccgt ggactccctg gagtcacccc atggctacct
     2341 gtcagacgtg gcctcgccgc cactgctgcc ctccccgttc cagcagtctc cgtccgtgcc
     2401 cctcaaccac ctgcctggga tgcccgacac ccacctgggc atcgggcacc tgaacgtggc
     2461 ggccaagccc gagatggcgg cgctgggtgg gggcggccgg ctggcctttg agactggccc
     2521 acctcgtctc tcccacctgc ctgtggcctc tggcaccagc accgtcctgg gctccagcag
     2581 cggaggggcc ctgaatttca ctgtgggcgg gtccaccagt ttgaatggtc aatgcgagtg
     2641 gctgtcccgg ctgcagagcg gcatggtgcc gaaccaatac aaccctctgc gggggagtgt
     2701 ggcaccaggc cccctgagca cacaggcccc ctccctgcag catggcatgg taggcccgct
     2761 gcacagtagc cttgctgcca gcgccctgtc ccagatgatg agctaccagg gcctgcccag
     2821 cacccggctg gccacccagc ctcacctggt gcagacccag caggtgcagc cacaaaactt
     2881 acagatgcag cagcagaacc tgcagccagc aaacatccag cagcagcaaa gcctgcagcc
     2941 gccaccacca ccaccacagc cgcaccttgg cgtgagctca gcagccagcg gccacctggg
     3001 ccggagcttc ctgagtggag agccgagcca ggcagacgtg cagccactgg gccccagcag
     3061 cctggcggtg cacactattc tgccccagga gagccccgcc ctgcccacgt cgctgccatc
     3121 ctcgctggtc ccacccgtga ccgcagccca gttcctgacg cccccctcgc agcacagcta
     3181 ctcctcgcct gtggacaaca cccccagcca ccagctacag gtgcctgagc accccttcct
     3241 caccccgtcc cctgagtccc ctgaccagtg gtccagctcg tccccgcatt ccaacgtctc
     3301 cgactggtcc gagggcgtct ccagccctcc caccagcatg cagtcccaga tcgcccgcat
     3361 tccggaggcc ttcaagtaaa agggtgggcg cgccgaccca gctttcttgt acaaagtggt
     3421 gatctagcta aaacgcggcc tcgaatcgaa ttccgccccc ccccctctcc ctcccccccc
     3481 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
     3541 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     3601 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     3661 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
     3721 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     3781 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     3841 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     3901 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
     3961 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     4021 aaaacacgat gataatatgg ccacaaccat ggtgagcaag ggcgaggagc tgttcaccgg
     4081 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     4141 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     4201 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     4261 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     4321 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     4381 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     4441 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     4501 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     4561 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     4621 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     4681 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     4741 tctcggcatg gacgagctgt acaagtaaag cggccgcgat ctttttccct ctgccaaaaa
     4801 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
     4861 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     4921 ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca
     4981 tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc
     5041 ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat
     5101 attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact
     5161 agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg
     5221 gagatccctc gacctgcagg aattcgatta cttaagccta ggatccagat ctgtacattc
     5281 gaatcgatga attcgttaac catggaagct tctgcagcgg ccgcactagc ggtggcggcc
     5341 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     5401 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     5461 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     5521 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     5581 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     5641 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     5701 attagtcatc gctattacca tgatgatgcg gttttggcag tacatcaatg ggcgtggata
     5761 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     5821 ttgactagtg gagccgagag taattcatac aaaaggaggg atcgccttcg caaggggaga
     5881 gcccagggac cgtccctaaa ttctcacaga cccaaatccc tgtagccgcc ccacgacagc
     5941 gcgaggagca tgcgcccagg gctgagcgcg ggtagatcag agcacacaag ctcacagtcc
     6001 ccggcggtgg ggggaggggc gcgctgagcg ggggccaggg agctggcgcg gggcaaactg
     6061 ggaaagtggt gtcgtgtgct ggctccgccc tcttcccgag ggtgggggag aacggtatat
     6121 aagtgcggta gtcgccttgg acgttctttt tcgcaacggg tttgccgtca gaacgcaggt
     6181 gagtggcggg tgtggcttcc gcgggccccg gagctggagc cctgctctga gcgggccggg
     6241 ctgatatgcg agtgtcgtcc gcagggttta gctgtgagca ttcccacttc gagtggcggg
     6301 cggtgcgggg gtgagagtgc gaggcctagc ggcaaccccg tagcctcgcc tcgtgtccgg
     6361 cttgaggcct agcgtggtgt ccgccgccgc gtgccactcc ggccgcacta tgcgtttttt
     6421 gtccttgctg ccctcgattg ccttccagca gcatgggcta acaaagggag ggtgtggggc
     6481 tcactcttaa ggagcccatg aagcttacgt tggataggaa tggaagggca ggaggggcga
     6541 ctggggcccg cccgccttcg gagcacatgt ccgacgccac ctggatgggg cgaggcctgt
     6601 ggctttccga agcaatcggg cgtgagttta gcctacctgg gccatgtggc cctagcactg
     6661 ggcacggtct ggcctggcgg tgccgcgttc ccttgcctcc caacaagggt gaggccgtcc
     6721 cgcccggcac cagttgcttg cgcggaaaga tggccgctcc cggggccctg ttgcaaggag
     6781 ctcaaaatgg aggacgcggc agcccggtgg agcgggcggg tgagtcaccc acacaaagga
     6841 agagggcctt gcccctcgcc ggccgctgct tcctgtgacc ccgtggtcta tcggccgcat
     6901 agtcacctcg ggcttctctt gagcaccgct cgtcgcggcg gggggagggg atctaatggc
     6961 gttggagttt gttcacattt ggtgggtgga gactagtcag gccagcctgg cgctggaagt
     7021 cattcttgga atttgcccct ttgagtttgg agcgaggcta attctcaagc ctcttagcgg
     7081 ttcaaaggta ttttctaaac ccgtttccag gtgttgtgaa agccaccgct aattcaaagc
     7141 aatccggagt aattcaccat gtctagactg gacaagagca aagtcataaa cggcgctctg
     7201 gaattactca atggagtcgg tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg
     7261 ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct gctcgatgcc
     7321 ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga aggcgagtca
     7381 tggcaagact ttctgcggaa caacgccaag tcattccgct gtgctctcct ctcacatcgc
     7441 gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga aaccctggaa
     7501 aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact gtacgctctg
     7561 tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca tcaagtagca
     7621 aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag acaagcaatt
     7681 gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga actaatcata
     7741 tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cggccgacgc ccttgacgat
     7801 tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga tatgctgcct
     7861 gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaactaag taaggatcat
     7921 tccgcccccc ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa
     7981 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     8041 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     8101 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     8161 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     8221 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     8281 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     8341 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     8401 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     8461 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     8521 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     8581 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     8641 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     8701 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     8761 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     8821 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     8881 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     8941 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     9001 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     9061 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
     9121 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgaaatg
     9181 accgaccaag cgaattcgct aggattatcc ctaatacctg ccaccccact cttaatcagt
     9241 ggtggaagaa cggtctcaga actgtttgtt tcaattggcc atttaagttt agtagtaaaa
     9301 gactggttaa tgataacaat gcatcgtaaa accttcagaa ggaaaggaga atgttttgtg
     9361 gaccactttg gttttctttt ttgcgtgtgg cagttttaag ttattagttt ttaaaatcag
     9421 tactttttaa tggaaacaac ttgaccaaaa atttgtcaca gaattttgag acccattaaa
     9481 aaagttaaat gagaaacctg tgtgttcctt tggtcaacac cgagacattt aggtgaaaga
     9541 catctaattc tggttttacg aatctggaaa cttcttgaaa atgtaattct tgagttaaca
     9601 cttctgggtg gagaataggg ttgttttccc cccacataat tggaagggga aggaatatca
     9661 tttaaagcta tgggagggtt gctttgatta caacactgga gagaaatgca gcatgttgct
     9721 gattgcctgt cactaaaaca ggccaaaaac tgagtccttg ggttgcatag aaagctgcca
     9781 aacgctagcc aacaagctcg tcatcgcttt gcagaagagc agagaggata tgctcatcgt
     9841 ctaaagaact acccatttta ttatatatta gtcacgatat ctataacaag aaaatatata
     9901 tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa
     9961 atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt
    10021 tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga
    10081 ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt
    10141 tcaagaatgc atgcgtcaat tttacgcaga ctatctttct agggttaaaa aagatttgcg
    10201 ctttactcga cctaaacttt aaacacgtca tagaatcttc gtttgacaaa aaccacattg
    10261 tggccaagct gtgtgacgcg acgcgcgcta aagaatggca aaccaagtcg cgcgaggtac
    10321 cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat ggtcatagct
    10381 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    10441 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    10501 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    10561 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    10621 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    10681 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    10741 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    10801 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    10861 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    10921 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    10981 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    11041 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    11101 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    11161 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    11221 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    11281 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    11341 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    11401 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    11461 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    11521 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    11581 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    11641 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    11701 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    11761 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    11821 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    11881 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    11941 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    12001 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    12061 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    12121 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    12181 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    12241 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    12301 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    12361 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    12421 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    12481 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
    12541 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
    12601 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
    12661 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
    12721 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
    12781 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
    12841 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
    12901 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
    12961 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
    13021 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
    13081 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
    13141 ttg
//
