LOCUS       sequence-458207-       11026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A Piggybac compatible, tet-on, NEO selectable, vector for expressing
            luciferase followed by a P2A cleavage peptide and H2A fused mCherry.
            .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11026)
  AUTHORS   Gillespie WB 3rd, Zhang Y, Ruiz OE, Cerda J 3rd, Ortiz-Guzman J, 
            Sherman M, Turner WD, Largoza G, Mosser LE, Fujimoto E, Chien CB, 
            Kwan KM, Arenkiel BR, Devine WP, Wythe JD
  TITLE     MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox for use in vertebrate model systems.
  JOURNAL   Development. 2025 Nov 15;152(22):dev204308. doi: 10.1242/dev.204308.
            Epub 2025 Nov 13.
  PUBMED    41230856
REFERENCE   2  (bases 1 to 11026)
  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: 
            "Development."; date: "2025-11-15"; volume: "152(22):dev204308. 
            doi"; pages: " 10.1242/dev.204308. Epub 2025 Nov 13"
COMMENT     Sequence Label: pB-Tet-On-Luciferase-P2A-H2A-mCherry (JDW 945)
FEATURES             Location/Qualifiers
     source          1..11026
                     /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"
     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    1542..1566
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1579..1588
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1585..3234
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1651..1669)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             3244..3300
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3697..4407
                     /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(3842..3860)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4110..4129
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4304..4327
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(4425..4449)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4564..4583)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4629..4684
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4683..4702)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     misc_feature    5067..5158
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5107..5126
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5187..5398
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(5550..6167,6168..6173,6174..6209,6210..6212,6213..6248,
                     6249..6251,6252..6287,6288..6296)
                     /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    6323..6767
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        6800..6846
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        6838..6856
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6866..7660
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6866..7660
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6920..6939)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7530..7549
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7707..8279
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   8653..8687
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(8836..8854)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8875..8891)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8875..8891)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8888..8910)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8899..8915
                     /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(8923..8929,8930..8947,8948..8953))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8968..8989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9106..9123)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9277..9865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9357..9376)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10036..10827,10828..10896))
                     /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     10659..10678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10897..11001)
                     /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 gtggcggcat cacaagtttg tacaaaaaag
     1561 caggctggat ccactagtgc caccatggaa gacgccaaaa acataaagaa aggcccggcg
     1621 ccattctatc cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga
     1681 tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact
     1741 tacgctgagt acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg
     1801 aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg
     1861 ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt
     1921 gaattgctca acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg
     1981 ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg
     2041 gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta
     2101 cctcccggtt ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt
     2161 gcactgatca tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat
     2221 agaactgcct gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt
     2281 ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca
     2341 ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg
     2401 tttctgagga gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc
     2461 tccttcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt
     2521 gcttctggtg gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat
     2581 ctgccaggta tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca
     2641 cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag
     2701 gttgtggatc tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg
     2761 agaggtccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt
     2821 gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc
     2881 ttcatcgttg accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct
     2941 gaattggaat ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt
     3001 cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg
     3061 atgacggaaa aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg
     3121 cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca
     3181 agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgggtacc
     3241 ggagccacta acttctccct gttgaaacaa gcaggggatg tcgaagagaa tcccgggcca
     3301 cctaggatgg caggtggaaa agcaggtaaa gacagtggaa aagccaagac gaaagccgtc
     3361 tccagatctc agagagctgg tctgcagttc cctgtaggtc gtatccacag gcacctgaag
     3421 acacgaacca ccagccatgg ccgtgtcgga gctacagctg cggtttacag tgctgctatc
     3481 ctggagtatc tcaccgctga ggtgctggag ttggctggaa acgcttctaa ggatctgaag
     3541 gtgaagcgaa tcactcccag acatttacag ctggccatcc gcggagacga ggagctggac
     3601 tcgcttatca aggccaccat tgctggagga ggagtcattc ctcacatcca caagtctctg
     3661 attggtaaga agggccagca gaaaaccgca ggggggatgg tgagcaaggg cgaggaggac
     3721 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     3781 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     3841 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtcccct
     3901 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     3961 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     4021 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     4081 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     4141 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     4201 aggctgaagc tgaaggacgg cggccactac gacgccgagg tcaagaccac ctacaaggcc
     4261 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagctgga catcacctcc
     4321 cacaacgagg actacaccat cgtggaacag tacgagcgcg ccgagggccg ccactccacc
     4381 ggcggcatgg acgagctgta caagtaatga tagctcgaga gatctcccag ctttcttgta
     4441 caaagtggtg atggccgctc tagcgaaggg atcccccatc gaattcactc ctcaggtgca
     4501 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     4561 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     4621 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     4681 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     4741 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     4801 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     4861 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     4921 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     4981 catagctgtc cctcttctct tatggagatc cctcgacctg caggaattcg attacttaag
     5041 cctaggatcc gtacattcga atcgataaca tacgctctcc atcaaaacaa aacgaaacaa
     5101 aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctgc
     5161 gatcgcgggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     5221 gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg
     5281 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     5341 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagac
     5401 tagtgggagc cgccaccatg agccatggct tcccgccgga ggtggaggag caggatgatg
     5461 gcacgctgcc catgtcttgt gcccaggaga gcgggatgga ccgtcaccct gcagcctgtg
     5521 cttctgctag gatcaatgtg gaattcacca tgtctagact ggacaagagc aaagtcataa
     5581 acggcgctct ggaattactc aatggagtcg gtatcgaagg cctgacgaca aggaaactcg
     5641 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     5701 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg
     5761 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcattccgc tgtgctctcc
     5821 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     5881 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     5941 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     6001 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     6061 gacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     6121 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccggccgacg
     6181 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     6241 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaactaa
     6301 gtaaggatcc cgtacgccta ggagcaggtt tccccaatga cacaaaacgt gcaacttgaa
     6361 actccgcctg gtctttccag gtctagaggg gtaacacttt gtactgcgtt tggctccacg
     6421 ctcgatccac tggcgagtgt tagtaacagc actgttgctt cgtagcggag catgacggcc
     6481 gtgggaactc ctccttggta acaaggaccc acggggccaa aagccacgcc cacacgggcc
     6541 cgtcatgtgt gcaaccccag cacggcgact ttactgcgaa acccacttta aagtgacatt
     6601 gaaactggta cccacacact ggtgacaggc taaggatgcc cttcaggtac cccgaggtaa
     6661 cacgcgacac tcgggatctg agaaggggac tggggcttct ataaaagcgc tcggtttaaa
     6721 aagcttctat gcctgaatag gtgaccggag gtcggcacct ttcctttgca attactgacc
     6781 ctatgaatac actgactgtt tgacaattaa tcatcggcat agtatatcgg catagtataa
     6841 tacgactcac tataggaggg ccaccatgat tgaacaagat ggattgcacg caggttctcc
     6901 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     6961 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     7021 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     7081 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     7141 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     7201 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     7261 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     7321 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     7381 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacacatg gcgatgcctg
     7441 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     7501 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     7561 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     7621 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     7681 atgaccgacc aagcgaattc gctaggatta tccctaatac ctgccacccc actcttaatc
     7741 agtggtggaa gaacggtctc agaactgttt gtttcaattg gccatttaag tttagtagta
     7801 aaagactggt taatgataac aatgcatcgt aaaaccttca gaaggaaagg agaatgtttt
     7861 gtggaccact ttggttttct tttttgcgtg tggcagtttt aagttattag tttttaaaat
     7921 cagtactttt taatggaaac aacttgacca aaaatttgtc acagaatttt gagacccatt
     7981 aaaaaagtta aatgagaaac ctgtgtgttc ctttggtcaa caccgagaca tttaggtgaa
     8041 agacatctaa ttctggtttt acgaatctgg aaacttcttg aaaatgtaat tcttgagtta
     8101 acacttctgg gtggagaata gggttgtttt ccccccacat aattggaagg ggaaggaata
     8161 tcatttaaag ctatgggagg gttgctttga ttacaacact ggagagaaat gcagcatgtt
     8221 gctgattgcc tgtcactaaa acaggccaaa aactgagtcc ttgggttgca tagaaagctg
     8281 ccaaacgcta gccaacaagc tcgtcatcgc tttgcagaag agcagagagg atatgctcat
     8341 cgtctaaaga actacccatt ttattatata ttagtcacga tatctataac aagaaaatat
     8401 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
     8461 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
     8521 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
     8581 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
     8641 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta aaaaagattt
     8701 gcgctttact cgacctaaac tttaaacacg tcatagaatc ttcgtttgac aaaaaccaca
     8761 ttgtggccaa gctgtgtgac gcgacgcgcg ctaaagaatg gcaaaccaag tcgcgcgagg
     8821 taccagcttt tgttcccttt agtgagggtt aatttcgagc ttggcgtaat catggtcata
     8881 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     8941 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     9001 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     9061 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     9121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     9181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     9241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     9301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     9361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     9421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     9481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     9541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     9601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     9661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     9721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     9781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     9841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     9901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     9961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    10021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    10081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    10141 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    10201 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    10261 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    10321 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    10381 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    10441 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    10501 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    10561 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    10621 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    10681 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    10741 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    10801 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    10861 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    10921 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    10981 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacct
//
