LOCUS       sequence-460713-       12419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A Piggybac compatible, tet-on, dual reporter encoding luciferase 
            followed by a P2A cleavage peptide and then an H2A fused mCherry for
            nuclear labeling..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12419)
  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 12419)
  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 1154)
FEATURES             Location/Qualifiers
     source          1..12419
                     /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"
     enhancer        5127..5506
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5621..6926
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    5963..6894
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(6943..7560,7561..7566,7567..7602,7603..7605,7606..7641,
                     7642..7644,7645..7680,7681..7689)
                     /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    7716..8160
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        8193..8239
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        8231..8249
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8259..9053
                     /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             8259..9053
                     /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(8313..8332)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8923..8942
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9100..9672
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   10046..10080
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(10229..10247)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10268..10284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10268..10284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10281..10303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10292..10308
                     /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(10316..10322,10323..10340,10341..10346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10361..10382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10499..10516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10670..11258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10750..10769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11429..12220,12221..12289))
                     /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     12052..12071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12290..12394)
                     /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 agatctgtac attcgaatcg atgaattcgt taaccatgga agcttctgca
     5101 gcggccgcac tagcggtggc ggcctcgaca ttgattattg actagttatt aatagtaatc
     5161 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     5221 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     5281 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     5341 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     5401 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     5461 tcctacttgg cagtacatct acgtattagt catcgctatt accatgatga tgcggttttg
     5521 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     5581 cattgacgtc aatgggagtt tgttttgact agtggagccg agagtaattc atacaaaagg
     5641 agggatcgcc ttcgcaaggg gagagcccag ggaccgtccc taaattctca cagacccaaa
     5701 tccctgtagc cgccccacga cagcgcgagg agcatgcgcc cagggctgag cgcgggtaga
     5761 tcagagcaca caagctcaca gtccccggcg gtggggggag gggcgcgctg agcgggggcc
     5821 agggagctgg cgcggggcaa actgggaaag tggtgtcgtg tgctggctcc gccctcttcc
     5881 cgagggtggg ggagaacggt atataagtgc ggtagtcgcc ttggacgttc tttttcgcaa
     5941 cgggtttgcc gtcagaacgc aggtgagtgg cgggtgtggc ttccgcgggc cccggagctg
     6001 gagccctgct ctgagcgggc cgggctgata tgcgagtgtc gtccgcaggg tttagctgtg
     6061 agcattccca cttcgagtgg cgggcggtgc gggggtgaga gtgcgaggcc tagcggcaac
     6121 cccgtagcct cgcctcgtgt ccggcttgag gcctagcgtg gtgtccgccg ccgcgtgcca
     6181 ctccggccgc actatgcgtt ttttgtcctt gctgccctcg attgccttcc agcagcatgg
     6241 gctaacaaag ggagggtgtg gggctcactc ttaaggagcc catgaagctt acgttggata
     6301 ggaatggaag ggcaggaggg gcgactgggg cccgcccgcc ttcggagcac atgtccgacg
     6361 ccacctggat ggggcgaggc ctgtggcttt ccgaagcaat cgggcgtgag tttagcctac
     6421 ctgggccatg tggccctagc actgggcacg gtctggcctg gcggtgccgc gttcccttgc
     6481 ctcccaacaa gggtgaggcc gtcccgcccg gcaccagttg cttgcgcgga aagatggccg
     6541 ctcccggggc cctgttgcaa ggagctcaaa atggaggacg cggcagcccg gtggagcggg
     6601 cgggtgagtc acccacacaa aggaagaggg ccttgcccct cgccggccgc tgcttcctgt
     6661 gaccccgtgg tctatcggcc gcatagtcac ctcgggcttc tcttgagcac cgctcgtcgc
     6721 ggcgggggga ggggatctaa tggcgttgga gtttgttcac atttggtggg tggagactag
     6781 tcaggccagc ctggcgctgg aagtcattct tggaatttgc ccctttgagt ttggagcgag
     6841 gctaattctc aagcctctta gcggttcaaa ggtattttct aaacccgttt ccaggtgttg
     6901 tgaaagccac cgctaattca aagcaatccg gagtaattca ccatgtctag actggacaag
     6961 agcaaagtca taaacggcgc tctggaatta ctcaatggag tcggtatcga aggcctgacg
     7021 acaaggaaac tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag
     7081 aacaagcggg ccctgctcga tgccctgcca atcgagatgc tggacaggca tcatacccac
     7141 ttctgccccc tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcattc
     7201 cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca
     7261 gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc
     7321 ctggagaacg cactgtacgc tctgtccgcc gtgggccact ttacactggg ctgcgtattg
     7381 gaggaacagg agcatcaagt agcaaaagag gaaagagaga cacctaccac cgattctatg
     7441 cccccacttc tgagacaagc aattgagctg ttcgaccggc agggagccga acctgccttc
     7501 cttttcggcc tggaactaat catatgtggc ctggagaaac agctaaagtg cgaaagcggc
     7561 gggccggccg acgcccttga cgattttgac ttagacatgc tcccagccga tgcccttgac
     7621 gactttgacc ttgatatgct gcctgctgac gctcttgacg attttgacct tgacatgctc
     7681 cccgggtaac taagtaagga tcccgtacgc ctaggagcag gtttccccaa tgacacaaaa
     7741 cgtgcaactt gaaactccgc ctggtctttc caggtctaga ggggtaacac tttgtactgc
     7801 gtttggctcc acgctcgatc cactggcgag tgttagtaac agcactgttg cttcgtagcg
     7861 gagcatgacg gccgtgggaa ctcctccttg gtaacaagga cccacggggc caaaagccac
     7921 gcccacacgg gcccgtcatg tgtgcaaccc cagcacggcg actttactgc gaaacccact
     7981 ttaaagtgac attgaaactg gtacccacac actggtgaca ggctaaggat gcccttcagg
     8041 taccccgagg taacacgcga cactcgggat ctgagaaggg gactggggct tctataaaag
     8101 cgctcggttt aaaaagcttc tatgcctgaa taggtgaccg gaggtcggca cctttccttt
     8161 gcaattactg accctatgaa tacactgact gtttgacaat taatcatcgg catagtatat
     8221 cggcatagta taatacgact cactatagga gggccaccat gattgaacaa gatggattgc
     8281 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     8341 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     8401 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     8461 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     8521 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     8581 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     8641 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     8701 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     8761 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgacac
     8821 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     8881 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     8941 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     9001 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     9061 tctggggttc gaaatgaccg accaagcgaa ttcgctagga ttatccctaa tacctgccac
     9121 cccactctta atcagtggtg gaagaacggt ctcagaactg tttgtttcaa ttggccattt
     9181 aagtttagta gtaaaagact ggttaatgat aacaatgcat cgtaaaacct tcagaaggaa
     9241 aggagaatgt tttgtggacc actttggttt tcttttttgc gtgtggcagt tttaagttat
     9301 tagtttttaa aatcagtact ttttaatgga aacaacttga ccaaaaattt gtcacagaat
     9361 tttgagaccc attaaaaaag ttaaatgaga aacctgtgtg ttcctttggt caacaccgag
     9421 acatttaggt gaaagacatc taattctggt tttacgaatc tggaaacttc ttgaaaatgt
     9481 aattcttgag ttaacacttc tgggtggaga atagggttgt tttcccccca cataattgga
     9541 aggggaagga atatcattta aagctatggg agggttgctt tgattacaac actggagaga
     9601 aatgcagcat gttgctgatt gcctgtcact aaaacaggcc aaaaactgag tccttgggtt
     9661 gcatagaaag ctgccaaacg ctagccaaca agctcgtcat cgctttgcag aagagcagag
     9721 aggatatgct catcgtctaa agaactaccc attttattat atattagtca cgatatctat
     9781 aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat aacaatataa
     9841 ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc attttgactc
     9901 acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca cgcctcacgg
     9961 gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc gctatttaga
    10021 aagagagagc aatatttcaa gaatgcatgc gtcaatttta cgcagactat ctttctaggg
    10081 ttaaaaaaga tttgcgcttt actcgaccta aactttaaac acgtcataga atcttcgttt
    10141 gacaaaaacc acattgtggc caagctgtgt gacgcgacgc gcgctaaaga atggcaaacc
    10201 aagtcgcgcg aggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
    10261 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
    10321 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
    10381 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
    10441 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
    10501 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
    10561 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
    10621 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
    10681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
    10741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
    10801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
    10861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
    10921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
    10981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
    11041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
    11101 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
    11161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
    11221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
    11281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
    11341 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
    11401 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
    11461 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
    11521 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
    11581 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
    11641 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
    11701 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
    11761 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
    11821 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
    11881 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
    11941 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
    12001 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
    12061 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
    12121 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
    12181 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
    12241 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
    12301 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
    12361 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacct
//
