LOCUS       sequence-487089-        9829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac transposon flanked, Tet-on, gateway destination vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9829)
  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 9829)
  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-EFS-mODC-rtTA-IRES-NEO (JDW 931)
FEATURES             Location/Qualifiers
     source          1..9829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..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     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   711..773
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1145..1163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1182..1200
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1219..1237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1256..1274
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1293..1311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1330..1348
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1367..1385
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1404..1422
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1444..1468
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1548..1672
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1697..1702,1703..1720,1721..1727)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1781..2440
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1847..1866)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2782..3087
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2966..2985
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3128..3252)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3367..3386)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3432..3487
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3486..3505)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     misc_feature    3868..3959
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3908..3927
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3988..4199
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(4351..4968,4969..4974,4975..5010,5011..5013,5014..5049,
                     5050..5052,5053..5088,5089..5097)
                     /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    5124..5568
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        5601..5647
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5639..5657
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5667..6461
                     /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             5667..6461
                     /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(5721..5740)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6331..6350
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6508..7080
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   7454..7488
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(7638..7656)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7677..7693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7677..7693)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7690..7712)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7701..7717
                     /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(7725..7731,7732..7749,7750..7755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7770..7791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7908..7925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8079..8667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8159..8178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8838..9629,9630..9698))
                     /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     9461..9480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9699..9803)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggaaattgta aacgttaata 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 gggcgcgtcg cgccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgg
      661 tattcacgac agcaggctga ataataaaaa aattagaaac tattatttaa ccctagaaag
      721 ataatcatat tgtgacgtac gttaaagata atcatgcgta aaattgacgc atgtgtttta
      781 tcggtctgta tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt
      841 acacttacat actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa
      901 aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa aacttttaaa cattctctct
      961 tttacaaaaa taaacttatt ttgtacttta aaaacagtca tgttgtatta taaaataagt
     1021 aattagctta acctatacat aatagaaaca aattatactt attagtcagt cagaaacaac
     1081 tttggcacat atcaatatta tgctctcgtt aattaactcg agggatccaa gctggccgca
     1141 cgtctcccta tcagtgatag agaagtcgac acgtctcgag ctccctatca gtgatagaga
     1201 aggtacgtct agaacgtctc cctatcagtg atagagaagt cgacacgtct cgagctccct
     1261 atcagtgata gagaaggtac gtctagaacg tctccctatc agtgatagag aagtcgacac
     1321 gtctcgagct ccctatcagt gatagagaag gtacgtctag aacgtctccc tatcagtgat
     1381 agagaagtcg acacgtctcg agctccctat cagtgataga gaaggtaccc cctatataag
     1441 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
     1501 ccatagaaga caccgggacc gatccagcct ccgcggtggc ggccatcaca agtttgtaca
     1561 aaaaagctga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
     1621 ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta tggcggccgc
     1681 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggattttgag
     1741 ttaggatccg tcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     1801 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     1861 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     1921 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     1981 gaatgctcat ccggaattcc gtatggcaat gaaagacggt gagctggtga tatgggatag
     2041 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     2101 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     2161 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     2221 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     2281 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     2341 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     2401 attacaacag tactgcgatg agtggcaggg cggggcgtaa acgcgtggat ccggcttact
     2461 aaaagccaga taacagtatg cgtatttgcg cgctgatttt tgcggtataa gaatatatac
     2521 tgatatgtat acccgaagta tgtcaaaaag aggtatgcta tgaagcagcg tattacagtg
     2581 acagttgaca gcgacagcta tcagttgctc aaggcatata tgatgtcaat atctccggtc
     2641 tggtaagcac aaccatgcag aatgaagccc gtcgtctgcg tgccgaacgc tggaaagcgg
     2701 aaaatcagga agggatggct gaggtcgccc ggtttattga aatgaacggc tcttttgctg
     2761 acgagaacag gggctggtga aatgcagttt aaggtttaca cctataaaag agagagccgt
     2821 tatcgtctgt ttgtggatgt acagagtgat attattgaca cgcccgggcg acggatggtg
     2881 atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg
     2941 gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc
     3001 tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc
     3061 attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag ccagtctgca
     3121 ggtcgaccat agtgactgga tatgttgtgt tttacagtat tatgtagtct gttttttatg
     3181 caaaatctaa tttaatatat tgatatttat atcattttac gtttctcgtt cagctttctt
     3241 gtacaaagtg gtgatggccg ctctagcgaa gggatccccc atcgaattca ctcctcaggt
     3301 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca
     3361 ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     3421 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     3481 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     3541 ggtttagagt ttggcaacat atgccatatg ctggctgcca tgaacaaagg tggctataaa
     3601 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct
     3661 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc
     3721 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag
     3781 tcatagctgt ccctcttctc ttatgaagat ccctcgacct gcaggaattc gattacttaa
     3841 gcctaggatc cgtacattcg aatcgataac atacgctctc catcaaaaca aaacgaaaca
     3901 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctctg
     3961 cgatcgcggg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     4021 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
     4081 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     4141 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacaga
     4201 ctagtgggag ccgccaccat gagccatggc ttcccgccgg aggtggagga gcaggatgat
     4261 ggcacgctgc ccatgtcttg tgcccaggag agcgggatgg accgtcaccc tgcagcctgt
     4321 gcttctgcta ggatcaatgt ggaattcacc atgtctagac tggacaagag caaagtcata
     4381 aacggcgctc tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc
     4441 gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc
     4501 ctgctcgatg ccctgccaat cgagatgctg gacaggcatc atacccactt ctgccccctg
     4561 gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcattccg ctgtgctctc
     4621 ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac
     4681 gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca
     4741 ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag
     4801 catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg
     4861 agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg
     4921 gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac
     4981 gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt
     5041 gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaacta
     5101 agtaaggatc ccgtacgcct aggagcaggt ttccccaatg acacaaaacg tgcaacttga
     5161 aactccgcct ggtctttcca ggtctagagg ggtaacactt tgtactgcgt ttggctccac
     5221 gctcgatcca ctggcgagtg ttagtaacag cactgttgct tcgtagcgga gcatgacggc
     5281 cgtgggaact cctccttggt aacaaggacc cacggggcca aaagccacgc ccacacgggc
     5341 ccgtcatgtg tgcaacccca gcacggcgac tttactgcga aacccacttt aaagtgacat
     5401 tgaaactggt acccacacac tggtgacagg ctaaggatgc ccttcaggta ccccgaggta
     5461 acacgcgaca ctcgggatct gagaagggga ctggggcttc tataaaagcg ctcggtttaa
     5521 aaagcttcta tgcctgaata ggtgaccgga ggtcggcacc tttcctttgc aattactgac
     5581 cctatgaata cactgactgt ttgacaatta atcatcggca tagtatatcg gcatagtata
     5641 atacgactca ctataggagg gccaccatga ttgaacaaga tggattgcac gcaggttctc
     5701 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5761 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5821 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5881 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5941 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     6001 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     6061 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     6121 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     6181 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacacat ggcgatgcct
     6241 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     6301 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6361 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6421 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6481 aatgaccgac caagcgaatt cgctaggatt atccctaata cctgccaccc cactcttaat
     6541 cagtggtgga agaacggtct cagaactgtt tgtttcaatt ggccatttaa gtttagtagt
     6601 aaaagactgg ttaatgataa caatgcatcg taaaaccttc agaaggaaag gagaatgttt
     6661 tgtggaccac tttggttttc ttttttgcgt gtggcagttt taagttatta gtttttaaaa
     6721 tcagtacttt ttaatggaaa caacttgacc aaaaatttgt cacagaattt tgagacccat
     6781 taaaaaagtt aaatgagaaa cctgtgtgtt cctttggtca acaccgagac atttaggtga
     6841 aagacatcta attctggttt tacgaatctg gaaacttctt gaaaatgtaa ttcttgagtt
     6901 aacacttctg ggtggagaat agggttgttt tccccccaca taattggaag gggaaggaat
     6961 atcatttaaa gctatgggag ggttgctttg attacaacac tggagagaaa tgcagcatgt
     7021 tgctgattgc ctgtcactaa aacaggccaa aaactgagtc cttgggttgc atagaaagct
     7081 gccaaacgct agccaacaag ctcgtcatcg ctttgcagaa gagcagagag gatatgctca
     7141 tcgtctaaag aactacccat tttattatat attagtcacg atatctataa caagaaaata
     7201 tatatataat aagttatcac gtaagtagaa catgaaataa caatataatt atcgtatgag
     7261 ttaaatctta aaagtcacgt aaaagataat catgcgtcat tttgactcac gcggtcgtta
     7321 tagttcaaaa tcagtgacac ttaccgcatt gacaagcacg cctcacggga gctccaagcg
     7381 gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa
     7441 tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt aaaaaagatt
     7501 tgcgctttac tcgacctaaa ctttaaacac gtcatagaat cttcgtttga caaaaaccac
     7561 attgtggcca agctgtgtga cgcgacgcgc gctaaagaat ggcaaaccaa gtcgcgcgag
     7621 gtacccagct tttgttccct ttagtgaggg ttaattccga gcttggcgta atcatggtca
     7681 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7741 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7801 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7861 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7921 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7981 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     8041 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     8101 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     8161 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     8221 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     8281 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     8341 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8401 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8461 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     8521 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8581 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8641 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8701 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8761 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8821 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8881 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8941 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     9001 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     9061 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     9121 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     9181 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9241 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9301 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9361 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9421 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9481 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9541 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9601 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9661 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9721 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9781 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
