LOCUS       sequence-366944-        8907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible mammalian cell expression of the AP4 helixCAM 
            and constitutive expression of an identifying fluorescent protein 
            (eBFP2). Flanked with PiggyBac sites for easy line construction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8907)
  AUTHORS   Chao G, Wannier TM, Gutierrez C, Borders NC, Appleton E, Chadha A, 
            Lebar T, Church GM
  TITLE     helixCAM: A platform for programmable cellular assembly in bacteria 
            and human cells.
  JOURNAL   Cell. 2022 Aug 23. pii: S0092-8674(22)01061-3. doi: 
            10.1016/j.cell.2022.08.012.
  PUBMED    36055250
REFERENCE   2  (bases 1 to 8907)
  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: "Cell. 2022 
            Aug 23. pii: S0092-8674(22)01061-3. doi: 
            10.1016/j.cell.2022.08.012."
COMMENT     Sequence Label: PB_TRE_AP4-PDGFR_EF1a_eBFP2
FEATURES             Location/Qualifiers
     source          1..8907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(162..266)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(292..747)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(429..450)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     641..660
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     874..896
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     888..905
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     889..905
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   1035..1097
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1546..1924
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1554..1572
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1590..1608
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1626..1644
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1662..1680
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1698..1716
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1734..1752
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1770..1788
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1976..2014
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    2059..2083
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     2083..2142
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2146..2172
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2281..2310
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2314..2460
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(2463..2487)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2492..2533
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2547..2564
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2621..2638)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2627..2834
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2911..3122
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3135..3403
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3437..4180
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             4181..4234
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4235..4237,4238..4240,4241..4951)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(4280..4301)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             4952..5005
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5006..5605
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5006..5025)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5502..5522
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5638..5759
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5675..5694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5729..5748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6522..6556
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7047..7063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7047..7063)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7060..7082)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7071..7087
                     /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(7095..7101,7102..7119,7120..7125))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7140..7161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7278..7295)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7449..8037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7529..7548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8208..8907,1..92,93..161))
                     /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     8831..8850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
       61 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
      121 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
      181 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
      241 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
      301 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
      361 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
      421 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
      481 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
      541 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
      601 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
      661 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
      721 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
      781 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
      841 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
      901 ccagtgagcg cgcctcgttc attcacgttt ttgaacccgt ggaggacggg cagactcgcg
      961 gtgcaaatgt gttttacagc gtgatggagc agatgaagat gctcgacacg ctgcagaaca
     1021 cgcagctaga ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg
     1081 tgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa
     1141 tagatattaa gttttattat atttacactt acatactaat aataaattca acaaacaatt
     1201 tatttatgtt tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa
     1261 caaaactttt atgagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc
     1321 ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg
     1381 catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg
     1441 ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg
     1501 aaaaggcctc cacggccact agtctttcgt cttcaagaat tcctcgagtt tactccctat
     1561 cagtgataga gaacgtatga agagtttact ccctatcagt gatagagaac gtatgcagac
     1621 tttactccct atcagtgata gagaacgtat aaggagttta ctccctatca gtgatagaga
     1681 acgtatgacc agtttactcc ctatcagtga tagagaacgt atctacagtt tactccctat
     1741 cagtgataga gaacgtatat ccagtttact ccctatcagt gatagagaac gtataagctt
     1801 taggcgtgta cggtgggcgc ctataaaagc agagctcgtt tagtgaaccg tcagatcgcc
     1861 tggagcaatt ccacaacact tttgtcttat accaactttc cgtaccactt cctaccctcg
     1921 taaaggtcta gagctagcga attcgaagcc accaactggc aggtgcgtat gtcgaggtag
     1981 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2041 gagaattggc tagcatcaac aagtttgtac aaaaaagttg gcatggagac agacacactc
     2101 ctgctatggg tactgctgct ctgggttcca ggttccactg gtgactatcc atatgatgtt
     2161 ccagattatg ctggggccag cccagaagat gagctggccg ctaacgagga agagttgcag
     2221 cagaacgaac agaaactggc gcaaatcaag caaaagttgc aggcgattaa gtatggagac
     2281 gaacaaaaac tcatctcaga agaggatctg aatgctgtgg gccaggacac gcaggaggtc
     2341 atcgtggtgc cacactcctt gccctttaag gtggtggtga tctcagccat cctggccctg
     2401 gtggtgctca ccatcatctc ccttatcatc ctcatcatgc tttggcagaa gaagccacgt
     2461 taacccaact ttcttgtaca aagtggttgc aggaaagcca atcccaaacc cactcctagg
     2521 tctcgacagt acagcataga ccggtccacc accaccacca ccactaagga tccggggttg
     2581 gggttgcgcc ttttccaagc acctgctaca tagcggtacc cctcgactgt gccttctagt
     2641 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2701 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2761 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2821 aggcatgctg gggatgcggt gggctcttcg aatttaaatc ggatccagct gaatctaagt
     2881 ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
     2941 agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa
     3001 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
     3061 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
     3121 agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc
     3181 gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg
     3241 cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc
     3301 cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac
     3361 tctacgtctt tgtttcgttt tctgttctgc gccgttacag atctaagctg tgaccggcgc
     3421 ctactctaga gccgccatgt ctagactgga caagagcaaa gtcataaact ctgctctgga
     3481 attactcaat ggagtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg
     3541 agttgagcag cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct
     3601 gccaatcgag atgctggaca ggcatcatac ccactcctgc cccctggaag gcgagtcatg
     3661 gcaagacttt ctgcggaaca acgccaagtc ataccgctgt gctctcctct cacatcgcga
     3721 cggggctaaa gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa
     3781 tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc
     3841 cgccgtgggc cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa
     3901 agaggaaaga gagacaccta ccaccgattc tatgccccca cttctgaaac aagcaattga
     3961 gctgttcgac cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg
     4021 tggcctggag aaacagctaa agtgcgaaag cggcgggccg accgacgccc ttgacgattt
     4081 tgacttagac atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc
     4141 tgacgctctt gacgattttg accttgacat gctccccggg gagggcagag gaagtcttct
     4201 aacatgcggt gacgtggagg agaatcccgg ccctatggtg agcaagggcg aggagctgtt
     4261 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     4321 cgtgaggggc gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg
     4381 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga gccacggcgt
     4441 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     4501 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca cctacaagac
     4561 ccgcgccgag gtgaagttcg agggcgacac cctagtgaac cgcatcgagc tgaagggcgt
     4621 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca
     4681 caacatctat atcatggccg tcaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4741 ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4801 cggcgacggc cccgtgctgc tgcccgacag ccactacctg agcacccagt ccgtgctgag
     4861 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttccgca ccgccgccgg
     4921 gatcactctc ggcatggacg agctgtacaa ggagggcaga ggaagtcttc taacatgcgg
     4981 tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc gcctcgccac
     5041 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     5101 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     5161 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     5221 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     5281 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5341 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5401 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5461 gcgcgccggg gtgccagcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5521 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5581 catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt gtgggtaaac
     5641 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5701 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5761 catgtctgga attgactcaa atgatgtcaa ttagtctatc agaagctcat ctggtctccc
     5821 ttccggggga caagacatcc ctgtttaata tttaaacagc agtgttccca aactgggttc
     5881 ttatatccct tgctctggtc aaccaggttg cagggtttcc tgtcctcaca ggaacgaagt
     5941 ccctaaagaa acagtggcag ccaggtttag ccccggaatt gactggattc cttttttagg
     6001 gcccattggt atggcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc
     6061 gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc
     6121 acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc
     6181 gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc
     6241 tgtccctgat atctataaca agaaaatata tatataataa gttatcacgt aagtagaaca
     6301 tgaaataaca atataattat cgtatgagtt aaatcttaaa agtcacgtaa aagataatca
     6361 tgcgtcattt tgactcacgc ggtcgttata gttcaaaatc agtgacactt accgcattga
     6421 caagcacgcc tcacgggagc tccaagcggc gactgagatg tcctaaatgc acagcgacgg
     6481 attcgcgcta tttagaaaga gagagcaata tttcaagaat gcatgcgtca attttacgca
     6541 gactatcttt ctagggttaa tctagctgca tcaggatcat atcgtcgggt cttttttccg
     6601 gctcagtcat cgcccaagct ggcgctatct gggcatcggg gaggaagaag cccgtgcctt
     6661 ttcccgcgag gttgaagcgg catggaaaga gtttgccgag gatgactgct gctgcattga
     6721 cgttgagcga aaacgcacgt ttaccatgat gattcgggaa ggtgtggcca tgcacgcctt
     6781 taacggtgaa ctgttcgttc aggccacctg ggataccagt tcgtcgcggc ttttccggac
     6841 acagttccgg atggtcagcc cgaagcgcat cagcaacccg aacaataccg gcgacagccg
     6901 gaactgccgt gccggtgtgc agattaatga cagcggtgcg gcgctgggat attacgtcag
     6961 cgaggacggg tatcctggct ggatgccgca gaaatggaca tggatacccc gtgagttacc
     7021 cggcgggcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     7081 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     7141 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7201 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7261 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     7321 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7381 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7441 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7501 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7561 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7621 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7681 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7741 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7801 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7861 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7921 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7981 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     8041 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     8101 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     8161 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     8221 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     8281 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     8341 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     8401 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8461 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8521 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8581 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8641 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8701 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8761 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8821 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8881 cgttcttcgg ggcgaaaact ctcaagg
//
