LOCUS       sequence-460485-        9501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A PiggyBac vector with a cre-dependent dual luciferase / nuclear 
            mCherry reporter. In the presence of cre, this tet-off vector will 
            be ubiquitously expressed in the absence of dox/tet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9501)
  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 9501)
  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-Off_FLEX_Luc-P2A-H2A-mCherry (JDW 970)
FEATURES             Location/Qualifiers
     source          1..9501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   660..694
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1016..1330
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    1024..1042
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1060..1078
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1095..1113
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1131..1149
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1167..1185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1202..1220
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1238..1256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1303..1327
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(1355..1388)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1439..1472)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1495..1519
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1537..2247)
                     /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(1617..1640)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1815..1834)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2084..2102
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(2644..2700)
                     /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             complement(2710..4359)
                     /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     4275..4293
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      4356..4365
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4378..4402)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    4428..4461
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    4512..4545
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4590..5178
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4643..4663)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5061..5072)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5211..5228)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5217..5328
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    5476..5524
                     /note="synthetic polyadenylation signal"
     misc_feature    5538..5629
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5578..5597
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5669..5880
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(6025..6642,6643..6648,6649..6684,6685..6687,6688..6723,
                     6724..6726,6727..6762,6763..6771)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(6784..6800)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6826..6844)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6870..7004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6907..6926)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6961..6980
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7227..7289
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(7313..7331)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7352..7368)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7352..7368)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7365..7387)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7376..7392
                     /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(7400..7406,7407..7424,7425..7430))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7445..7466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7583..7600)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7754..8342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7834..7853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8513..9304,9305..9373))
                     /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     9136..9155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9374..9478)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtaccttaac
      661 cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg ctctctcttt
      721 ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc ttggagctcc
      781 cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa cgaccgcgtg
      841 agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc atacgataat
      901 tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat tttcttgtta
      961 tagatatcgt cgacttaatt aacctaggat ttaaatgcgg ccgcacgcgc gtctcgagtt
     1021 tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac
     1081 gatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag
     1141 tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta tgtcgagttt
     1201 atccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacgt
     1261 atgtcgaggt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt tagtgaaccg
     1321 tcagatcgcc accggctcta gagtcgactc cggaataact tcgtatagga tactttatac
     1381 gaagttatgc agaatggtag ctggattgta gctgctatta gcaatatgaa acctcttaat
     1441 aacttcgtat agcatacatt atacgaagtt atggcgcgcc gctcgagagg atcaaccact
     1501 ttgtacaaga aagctgggag atctctcgag ctatcattac ttgtacagct cgtccatgcc
     1561 gccggtggag tggcggccct cggcgcgctc gtactgttcc acgatggtgt agtcctcgtt
     1621 gtgggaggtg atgtccagct tgatgttgac gttgtaggcg ccgggcagct gcacgggctt
     1681 cttggccttg taggtggtct tgacctcggc gtcgtagtgg ccgccgtcct tcagcttcag
     1741 cctctgcttg atctcgccct tcagggcgcc gtcctcgggg tacatccgct cggaggaggc
     1801 ctcccagccc atggtcttct tctgcattac ggggccgtcg gaggggaagt tggtgccgcg
     1861 cagcttcacc ttgtagatga actcgccgtc ctgcagggag gagtcctggg tcacggtcac
     1921 cacgccgccg tcctcgaagt tcatcacgcg ctcccacttg aagccctcgg ggaaggacag
     1981 cttcaagtag tcggggatgt cggcggggtg cttcacgtag gccttggagc cgtacatgaa
     2041 ctgaggggac aggatgtccc aggcgaaggg cagggggccg cccttggtca ccttcagctt
     2101 ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg ccctcgatct cgaactcgtg
     2161 gccgttcacg gagccctcca tgtgcacctt gaagcgcatg aactccttga tgatggccat
     2221 gttgtcctcc tcgcccttgc tcaccatccc ccctgcggtt ttctgctggc ccttcttacc
     2281 aatcagagac ttgtggatgt gaggaatgac tcctcctcca gcaatggtgg ccttgataag
     2341 cgagtccagc tcctcgtctc cgcggatggc cagctgtaaa tgtctgggag tgattcgctt
     2401 caccttcaga tccttagaag cgtttccagc caactccagc acctcagcgg tgagatactc
     2461 caggatagca gcactgtaaa ccgcagctgt agctccgaca cggccatggc tggtggttcg
     2521 tgtcttcagg tgcctgtgga tacgacctac agggaactgc agaccagctc tctgagatct
     2581 ggagacggct ttcgtcttgg cttttccact gtctttacct gcttttccac ctgccatcct
     2641 aggtggcccg ggattctctt cgacatcccc tgcttgtttc aacagggaga agttagtggc
     2701 tccggtaccc acggcgatct ttccgccctt cttggccttt atgaggatct ctctgatttt
     2761 tcttgcgtcg agttttccgg taagaccttt cggtacttcg tccacaaaca caactcctcc
     2821 gcgcaacttt ttcgcggttg ttacttgact ggcgacgtaa tccacgatct ctttttccgt
     2881 catcgtcttt ccgtgctcca aaacaacaac ggcggcggga agttcaccgg cgtcatcgtc
     2941 gggaagacct gcgacacctg cgtcgaagat gttggggtgt tggagcaaga tggattccaa
     3001 ttcagcggga gccacctgat agcctttgta cttaatcaga gacttcaggc ggtcaacgat
     3061 gaagaagtgt tcgtcttcgt cccagtaagc tatgtctcca gaatgtagcc atccatcctt
     3121 gtcaatcaag gcgttggtcg cttccggatt gtttacataa ccggacataa tcataggacc
     3181 tctcacacac agttcgcctc tttgattaac gcccagcgtt ttcccggtat ccagatccac
     3241 aaccttcgct tcaaaaaatg gaacaacttt accgaccgcg cccggtttat catccccctc
     3301 gggtgtaatc agaatagctg atgtagtctc agtgagccca tatccttgcc tgatacctgg
     3361 cagatggaac ctcttggcaa ccgcttcccc gacttcctta gagaggggag cgccaccaga
     3421 agcaatttcg tgtaaattag ataaatcgta tttgtcaatc agagtgcttt tggcgaagaa
     3481 ggagaatagg gttggcacca gcagcgcact ttgaatcttg taatcctgaa ggctcctcag
     3541 aaacagctct tcttcaaatc tatacattaa gacgactcga aatccacata tcaaatatcc
     3601 gagtgtagta aacattccaa aaccgtgatg gaatggaaca acacttaaaa tcgcagtatc
     3661 cggaatgatt tgattgccaa aaataggatc tctggcatgc gagaatctca cgcaggcagt
     3721 tctatgaggc agagcgacac ctttaggcag accagtagat ccagaggagt tcatgatcag
     3781 tgcaattgtc ttgtccctat cgaaggactc tggcacaaaa tcgtattcat taaaaccggg
     3841 aggtagatga gatgtgacga acgtgtacat cgactgaaat ccctggtaat ccgttttaga
     3901 atccatgata ataatttttt ggatgattgg gagctttttt tgcacgttca aaattttttg
     3961 caaccccttt ttggaaacga acaccacggt aggctgcgaa atgcccatac tgttgagcaa
     4021 ttcacgttca ttataaatgt cgttcgcggg cgcaactgca actccgataa ataacgcgcc
     4081 caacaccggc ataaagaatt gaagagagtt ttcactgcat acgacgattc tgtgatttgt
     4141 attcagccca tatcgtttca tagcttctgc caaccgaacg gacatttcga agtactcagc
     4201 gtaagtgatg tccacctcga tatgtgcatc tgtaaaagca attgttccag gaaccagggc
     4261 gtatctcttc atagccttat gcagttgctc tccagcggtt ccatcttcca gcggatagaa
     4321 tggcgccggg cctttcttta tgtttttggc gtcttccatg gtggcactag tggatccagc
     4381 ctgctttttt gtacaaactt gttgatcctt gaattcgaat cgctagcata acttcgtata
     4441 aagtatccta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa
     4501 tggtgcaaag aataacttcg tataatgtat gctatacgaa gttatggcgc cagcgatcca
     4561 ccggatctag ataactgatc cttatcgata atcaacctct ggattacaaa atttgtgaaa
     4621 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4681 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4741 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4801 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4861 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4921 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4981 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5041 cgcccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5101 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     5161 tttgggccgc ctccccgcat cgataccgtc gactagagct cgctgatcag cctcgactgt
     5221 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5281 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5341 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5401 agacaatagc agactagtac gtctctcaag gataagtaag taatattaag gtacgggagg
     5461 tattggacag gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg
     5521 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta
     5581 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctctg gcctaactgg
     5641 ccgacgcgtg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     5701 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
     5761 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     5821 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     5881 gggatccagc tcatgagcca tggcttcccg ccggaggtgg aggagcagga tgatggcacg
     5941 ctgcccatgt cttgtgccca ggagagcggg atggaccgtc accctgcagc ctgtgcttct
     6001 gctaggatca atgtggaatt caccatgtct agactggaca agagcaaagt cataaactct
     6061 gctctggaat tactcaatga agtcggtatc gaaggcctga cgacaaggaa actcgctcaa
     6121 aagctgggag ttgagcagcc taccctgtac tggcacgtga agaacaagcg ggccctgctc
     6181 gatgccctgg caatcgagat gctggacagg catcataccc acttctgccc cctggaaggc
     6241 gagtcatggc aagactttct gcggaacaac gccaagtcat tccgctgtgc tctcctctca
     6301 catcgcgacg gggctaaagt gcatctcggc acccgcccaa cagagaaaca gtacgaaacc
     6361 ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct ccctggagaa cgcactgtac
     6421 gctctgtccg ccgtgggcca ctttacactg ggctgcgtat tggaggatca ggagcatcaa
     6481 gtagcaaaag aggaaagaga gacacctacc accgattcta tgcccccact tctgagacaa
     6541 gcaattgagc tgttcgacca tcagggagcc gaacctgcct tccttttcgg cctggaacta
     6601 atcatatgtg gcctggagaa acagctaaag tgcgaaagcg gcgggccggc cgacgccctt
     6661 gacgattttg acttagacat gctcccagcc gatgcccttg acgactttga ccttgatatg
     6721 ctgcctgctg acgctcttga cgattttgac cttgacatgc tccccgggta actaagtaag
     6781 catcgatacc gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt
     6841 attacgcgcg cagcggccga ccatggccca acttgtttat tgcagcttat aatggttaca
     6901 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6961 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctccgga cacgtgcgga
     7021 ccgagcggcc gcaagcttgt ttaaacgtta acggatcctt tgttacttta tagaagaaat
     7081 tttgagtttt tgtttttttt taataaataa ataaacataa ataaattgtt tgttgaattt
     7141 attattagta tgtaagtgta aatataataa aacttaatat ctattcaaat taataaataa
     7201 acctcgatat acagaccgat aaaacacatg cgtcaatttt acgcatgatt atctttaacg
     7261 tacgtcacaa tatgattatc tttctagggt taaagagctc cagcttttgt tccctttagt
     7321 gagggttaat ttcgagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     7381 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7441 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7501 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7561 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7621 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7681 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8161 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8461 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8521 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8581 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8641 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8701 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8761 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8821 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8881 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8941 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     9001 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     9061 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     9121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     9181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9481 catttccccg aaaagtgcca c
//
