LOCUS       sequence-366950-        8898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible mammalian cell expression of the AP10 helixCAM
            and constitutive expression of an identifying fluorescent protein 
            (mOrange). Flanked with PiggyBac sites for easy line construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8898)
  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 8898)
  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_AP10-PDGFR_EF1a_mOrange
FEATURES             Location/Qualifiers
     source          1..8898
                     /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             4235..4942
                     /codon_start=1
                     /product="orange monomeric derivative of DsRed fluorescent 
                     protein (Shanet et al., 2004)"
                     /label=mOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     4648..4667
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4943..4996
                     /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             4997..5596
                     /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(4997..5016)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5493..5513
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5629..5750
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5666..5685)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5720..5739
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6513..6547
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7038..7054)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7038..7054)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7051..7073)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7062..7078
                     /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(7086..7092,7093..7110,7111..7116))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7131..7152
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7269..7286)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7440..8028)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7520..7539)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8199..8898,1..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     8822..8841
                     /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 tcctgaagat aagcttgcgc aaattaaaga aaaactgcag
     2221 cagattaaag aagaacttgc cgcgaatgaa gagaaactcc aggcgaacaa gtacggggac
     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 aggagaataa
     4261 catggccatc atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
     4321 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggct ttcagaccgc
     4381 taagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     4441 gttcacctac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttcaa
     4501 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     4561 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     4621 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     4681 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagatgag
     4741 gctgaagctg aaggacggcg gccactacac ctccgaggtc aagaccacct acaaggccaa
     4801 gaagcccgtg cagctgcccg gcgcctacat cgtcggcatc aagttggaca tcacctccca
     4861 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     4921 cggcatggac gagctgtaca aggagggcag aggaagtctt ctaacatgcg gtgacgtgga
     4981 ggagaatccc ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     5041 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     5101 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     5161 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     5221 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     5281 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     5341 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     5401 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     5461 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     5521 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     5581 caagcccggt gcctgaaatc aacctctgga ttacaaaatt tgtgggtaaa cttgtttatt
     5641 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5701 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5761 aattgactca aatgatgtca attagtctat cagaagctca tctggtctcc cttccggggg
     5821 acaagacatc cctgtttaat atttaaacag cagtgttccc aaactgggtt cttatatccc
     5881 ttgctctggt caaccaggtt gcagggtttc ctgtcctcac aggaacgaag tccctaaaga
     5941 aacagtggca gccaggttta gccccggaat tgactggatt ccttttttag ggcccattgg
     6001 tatggctttt tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt
     6061 tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg
     6121 gctcggggat gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc
     6181 ccctagcggg ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctga
     6241 tatctataac aagaaaatat atatataata agttatcacg taagtagaac atgaaataac
     6301 aatataatta tcgtatgagt taaatcttaa aagtcacgta aaagataatc atgcgtcatt
     6361 ttgactcacg cggtcgttat agttcaaaat cagtgacact taccgcattg acaagcacgc
     6421 ctcacgggag ctccaagcgg cgactgagat gtcctaaatg cacagcgacg gattcgcgct
     6481 atttagaaag agagagcaat atttcaagaa tgcatgcgtc aattttacgc agactatctt
     6541 tctagggtta atctagctgc atcaggatca tatcgtcggg tcttttttcc ggctcagtca
     6601 tcgcccaagc tggcgctatc tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga
     6661 ggttgaagcg gcatggaaag agtttgccga ggatgactgc tgctgcattg acgttgagcg
     6721 aaaacgcacg tttaccatga tgattcggga aggtgtggcc atgcacgcct ttaacggtga
     6781 actgttcgtt caggccacct gggataccag ttcgtcgcgg cttttccgga cacagttccg
     6841 gatggtcagc ccgaagcgca tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg
     6901 tgccggtgtg cagattaatg acagcggtgc ggcgctggga tattacgtca gcgaggacgg
     6961 gtatcctggc tggatgccgc agaaatggac atggataccc cgtgagttac ccggcgggcg
     7021 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     7081 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     7141 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     7201 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     7261 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     7321 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     7381 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     7441 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7501 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7561 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7621 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7681 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7741 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7801 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7861 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7921 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7981 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     8041 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     8101 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     8161 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     8221 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     8281 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     8341 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     8401 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     8461 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8521 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8581 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8641 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8701 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8761 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8821 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8881 gggcgaaaac tctcaagg
//
