LOCUS       sequence-460707-       12015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A CAGGS driven, piggybac compatible, tet-on expression vector 
            containing a luciferase reporter followed by a P2A cleavage peptide 
            and H2A fused mCherry for nuclear labeling. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12015)
  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 12015)
  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-TetOn-DEST-hEF1a-mODC-rtTA-IRES-NEO (JDW 1130)
FEATURES             Location/Qualifiers
     source          1..12015
                     /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        5194..6372
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5425..6363
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     6320..6340
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(6539..7156,7157..7162,7163..7198,7199..7201,7202..7237,
                     7238..7240,7241..7276,7277..7285)
                     /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    7312..7756
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        7789..7835
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        7827..7845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7855..8649
                     /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             7855..8649
                     /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(7909..7928)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8519..8538
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8696..9268
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   9642..9676
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(9825..9843)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9864..9880)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9864..9880)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9877..9899)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9888..9904
                     /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(9912..9918,9919..9936,9937..9942))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9957..9978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10095..10112)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10266..10854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10346..10365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11025..11816,11817..11885))
                     /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     11648..11667
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11886..11990)
                     /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 gatcgcttct aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag
     5221 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
     5281 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     5341 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     5401 aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc
     5461 tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt
     5521 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
     5581 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga
     5641 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
     5701 ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
     5761 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     5821 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     5881 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     5941 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     6001 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     6061 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     6121 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     6181 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     6241 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     6301 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     6361 caggtgtcgt gaggaattag cttggtaact agtgggagcc gccaccatga gccatggctt
     6421 cccgccggag gtggaggagc aggatgatgg cacgctgccc atgtcttgtg cccaggagag
     6481 cgggatggac cgtcaccctg cagcctgtgc ttctgctagg atcaatgtgg aattcaccat
     6541 gtctagactg gacaagagca aagtcataaa cggcgctctg gaattactca atggagtcgg
     6601 tatcgaaggc ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct
     6661 gtactggcac gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga
     6721 caggcatcat acccacttct gccccctgga aggcgagtca tggcaagact ttctgcggaa
     6781 caacgccaag tcattccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct
     6841 cggcacccgc ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg
     6901 tcagcaaggc ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac
     6961 actgggctgc gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc
     7021 taccaccgat tctatgcccc cacttctgag acaagcaatt gagctgttcg accggcaggg
     7081 agccgaacct gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct
     7141 aaagtgcgaa agcggcgggc cggccgacgc ccttgacgat tttgacttag acatgctccc
     7201 agccgatgcc cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt
     7261 tgaccttgac atgctccccg ggtaactaag taaggatccc gtacgcctag gagcaggttt
     7321 ccccaatgac acaaaacgtg caacttgaaa ctccgcctgg tctttccagg tctagagggg
     7381 taacactttg tactgcgttt ggctccacgc tcgatccact ggcgagtgtt agtaacagca
     7441 ctgttgcttc gtagcggagc atgacggccg tgggaactcc tccttggtaa caaggaccca
     7501 cggggccaaa agccacgccc acacgggccc gtcatgtgtg caaccccagc acggcgactt
     7561 tactgcgaaa cccactttaa agtgacattg aaactggtac ccacacactg gtgacaggct
     7621 aaggatgccc ttcaggtacc ccgaggtaac acgcgacact cgggatctga gaaggggact
     7681 ggggcttcta taaaagcgct cggtttaaaa agcttctatg cctgaatagg tgaccggagg
     7741 tcggcacctt tcctttgcaa ttactgaccc tatgaataca ctgactgttt gacaattaat
     7801 catcggcata gtatatcggc atagtataat acgactcact ataggagggc caccatgatt
     7861 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     7921 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     7981 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     8041 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     8101 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     8161 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     8221 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     8281 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     8341 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     8401 gatctcgtcg tgacacatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     8461 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     8521 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     8581 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     8641 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgaattcg ctaggattat
     8701 ccctaatacc tgccacccca ctcttaatca gtggtggaag aacggtctca gaactgtttg
     8761 tttcaattgg ccatttaagt ttagtagtaa aagactggtt aatgataaca atgcatcgta
     8821 aaaccttcag aaggaaagga gaatgttttg tggaccactt tggttttctt ttttgcgtgt
     8881 ggcagtttta agttattagt ttttaaaatc agtacttttt aatggaaaca acttgaccaa
     8941 aaatttgtca cagaattttg agacccatta aaaaagttaa atgagaaacc tgtgtgttcc
     9001 tttggtcaac accgagacat ttaggtgaaa gacatctaat tctggtttta cgaatctgga
     9061 aacttcttga aaatgtaatt cttgagttaa cacttctggg tggagaatag ggttgttttc
     9121 cccccacata attggaaggg gaaggaatat catttaaagc tatgggaggg ttgctttgat
     9181 tacaacactg gagagaaatg cagcatgttg ctgattgcct gtcactaaaa caggccaaaa
     9241 actgagtcct tgggttgcat agaaagctgc caaacgctag ccaacaagct cgtcatcgct
     9301 ttgcagaaga gcagagagga tatgctcatc gtctaaagaa ctacccattt tattatatat
     9361 tagtcacgat atctataaca agaaaatata tatataataa gttatcacgt aagtagaaca
     9421 tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa aagataatca
     9481 tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt accgcattga
     9541 caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc acagcgacgg
     9601 attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca attttacgca
     9661 gactatcttt ctagggttaa aaaagatttg cgctttactc gacctaaact ttaaacacgt
     9721 catagaatct tcgtttgaca aaaaccacat tgtggccaag ctgtgtgacg cgacgcgcgc
     9781 taaagaatgg caaaccaagt cgcgcgaggt accagctttt gttcccttta gtgagggtta
     9841 atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     9901 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     9961 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
    10021 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
    10081 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
    10141 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
    10201 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
    10261 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
    10321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
    10381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
    10441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
    10501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
    10561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
    10621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
    10681 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
    10741 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
    10801 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
    10861 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
    10921 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
    10981 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
    11041 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
    11101 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
    11161 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
    11221 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
    11281 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
    11341 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    11401 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    11461 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    11521 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
    11581 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    11641 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    11701 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    11761 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    11821 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    11881 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
    11941 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
    12001 cgaaaagtgc cacct
//
