LOCUS       sequence-460682-        8298 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A Piggybac-compatible, cre-dependent, Tet-off destination vector 
            with a downstream WPRE. EFS driving destabilized tTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8298)
  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 8298)
  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_DEST_EFS_mODC_tTA (JDW 936)
FEATURES             Location/Qualifiers
     source          1..8298
                     /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..1619
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1660..1965)
                     /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     complement(1762..1781)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(2307..2966)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2881..2900
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(3020..3026,3027..3044,3045..3050))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(3075..3199)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    3225..3258
                     /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    3309..3342
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3387..3975
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3440..3460)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3858..3869)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4008..4025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4014..4125
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4273..4321
                     /note="synthetic polyadenylation signal"
     misc_feature    4335..4426
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4375..4394
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        4466..4677
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(4822..5439,5440..5445,5446..5481,5482..5484,5485..5520,
                     5521..5523,5524..5559,5560..5568)
                     /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(5581..5597)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(5623..5641)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    5667..5801
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5704..5723)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5758..5777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6024..6086
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(6110..6128)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6149..6165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6149..6165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6162..6184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6173..6189
                     /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(6197..6203,6204..6221,6222..6227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6242..6263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6380..6397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6551..7139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6631..6650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7310..8101,8102..8170))
                     /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     7933..7952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8171..8275)
                     /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 aagctgaacg agaaacgtaa aatgatataa atatcaatat attaaattag
     1561 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatgg
     1621 tcgacctgca gactggctgt gtataaggga gcctgacatt tatattcccc agaacatcag
     1681 gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt cttccccgat
     1741 aacggagacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt catccccgat
     1801 atgcaccacc gggtaaagtt cacgggagac tttatctgac agcagacgtg cactggccag
     1861 ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat ccacaaacag
     1921 acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac cagcccctgt
     1981 tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc atcccttcct
     2041 gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc atggttgtgc
     2101 ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc tgtcgctgtc
     2161 aactgtcact gtaatacgct gcttcatagc atacctcttt ttgacatact tcgggtatac
     2221 atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata ctgttatctg
     2281 gcttttagta agccggatcc agatctttac gccccgccct gccactcatc gcagtactgt
     2341 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     2401 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     2461 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     2521 ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     2581 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     2641 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     2701 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa ttccggatga
     2761 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     2821 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     2881 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     2941 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgacgga
     3001 tcctaactca aaatccacac attatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3061 cctaatgcgg ccgccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt
     3121 ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag
     3181 cttttttgta caaacttgtt gatccttgaa ttcgaatcgc tagcataact tcgtataaag
     3241 tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc tttagaatgg
     3301 tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atggcgccag cgatccaccg
     3361 gatctagata actgatcctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat
     3421 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     3481 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     3541 ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca
     3601 ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt
     3661 ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg
     3721 cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga
     3781 aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgc
     3841 ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc
     3901 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3961 gggccgcctc cccgcatcga taccgtcgac tagagctcgc tgatcagcct cgactgtgcc
     4021 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     4081 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4141 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4201 caatagcaga ctagtacgtc tctcaaggat aagtaagtaa tattaaggta cgggaggtat
     4261 tggacaggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     4321 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     4381 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctggcc taactggccg
     4441 acgcgtggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4501 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     4561 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     4621 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagggg
     4681 atccagctca tgagccatgg cttcccgccg gaggtggagg agcaggatga tggcacgctg
     4741 cccatgtctt gtgcccagga gagcgggatg gaccgtcacc ctgcagcctg tgcttctgct
     4801 aggatcaatg tggaattcac catgtctaga ctggacaaga gcaaagtcat aaactctgct
     4861 ctggaattac tcaatgaagt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag
     4921 ctgggagttg agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat
     4981 gccctggcaa tcgagatgct ggacaggcat catacccact tctgccccct ggaaggcgag
     5041 tcatggcaag actttctgcg gaacaacgcc aagtcattcc gctgtgctct cctctcacat
     5101 cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg
     5161 gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct
     5221 ctgtccgccg tgggccactt tacactgggc tgcgtattgg aggatcagga gcatcaagta
     5281 gcaaaagagg aaagagagac acctaccacc gattctatgc ccccacttct gagacaagca
     5341 attgagctgt tcgaccatca gggagccgaa cctgccttcc ttttcggcct ggaactaatc
     5401 atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg ggccggccga cgcccttgac
     5461 gattttgact tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg
     5521 cctgctgacg ctcttgacga ttttgacctt gacatgctcc ccgggtaact aagtaagcat
     5581 cgataccgtc gacctcgagg gggggcccgg tacccaattc gccctatagt gagtcgtatt
     5641 acgcgcgcag cggccgacca tggcccaact tgtttattgc agcttataat ggttacaaat
     5701 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5761 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctccggacac gtgcggaccg
     5821 agcggccgca agcttgttta aacgttaacg gatcctttgt tactttatag aagaaatttt
     5881 gagtttttgt ttttttttaa taaataaata aacataaata aattgtttgt tgaatttatt
     5941 attagtatgt aagtgtaaat ataataaaac ttaatatcta ttcaaattaa taaataaacc
     6001 tcgatataca gaccgataaa acacatgcgt caattttacg catgattatc tttaacgtac
     6061 gtcacaatat gattatcttt ctagggttaa agagctccag cttttgttcc ctttagtgag
     6121 ggttaatttc gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6181 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6241 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6301 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6361 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6421 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6481 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6541 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6601 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6661 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6721 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6781 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6841 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6901 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6961 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7021 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7081 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7141 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7201 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7261 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7321 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7381 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7441 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7501 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7561 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7621 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7681 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7741 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7801 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7861 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7921 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7981 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8041 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8101 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8161 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8221 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8281 ttccccgaaa agtgccac
//
