LOCUS       sequence-366940-        8769 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible mammalian cell expression of the sg30 helixCAM
            and constitutive expression of an identifying fluorescent protein 
            (mCherry). Flanked with PiggyBac sites for easy line construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8769)
  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 8769)
  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_sg30-PDGFR_EF1a_mCherry
FEATURES             Location/Qualifiers
     source          1..8769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(25..129)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(155..610)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(292..313)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     504..523
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     737..759
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     751..768
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     752..768
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   898..960
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1409..1787
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1417..1435
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1453..1471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1489..1507
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1525..1543
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1561..1579
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1597..1615
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1633..1651
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1839..1877
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    1922..1946
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     1946..2005
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2015..2032
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2159..2188
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2192..2338
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(2349..2366)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2355..2562
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2639..2850
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2863..3131
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3165..3908
                     /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             3909..3962
                     /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             3972..4676
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGMVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(4114..4132)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4382..4401
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4677..4730
                     /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             4731..5330
                     /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(4731..4750)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5227..5247
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5363..5484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5400..5419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5454..5473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6247..6281
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6772..6788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6772..6788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6785..6807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6796..6812
                     /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(6820..6826,6827..6844,6845..6850))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6865..6886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7003..7020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7174..7762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7254..7273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7933..8724,8725..8769,1..24))
                     /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     8556..8575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
       61 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      121 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
      181 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
      241 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
      301 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
      361 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
      421 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
      481 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
      541 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
      601 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
      661 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
      721 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcctcg
      781 ttcattcacg tttttgaacc cgtggaggac gggcagactc gcggtgcaaa tgtgttttac
      841 agcgtgatgg agcagatgaa gatgctcgac acgctgcaga acacgcagct agattaaccc
      901 tagaaagata atcatattgt gacgtacgtt aaagataatc atgtgtaaaa ttgacgcatg
      961 tgttttatcg gtctgtatat cgaggtttat ttattaattt gaatagatat taagttttat
     1021 tatatttaca cttacatact aataataaat tcaacaaaca atttatttat gtttatttat
     1081 ttattaaaaa aaacaaaaac tcaaaatttc ttctataaag taacaaaact tttatgaggg
     1141 acagcccccc cccaaagccc ccagggatgt aattacgtcc ctcccccgct agggggcagc
     1201 agcgagccgc ccggggctcc gctccggtcc ggcgctcccc ccgcatcccc gagccggcag
     1261 cgtgcgggga cagcccgggc acggggaagg tggcacggga tcgctttcct ctgaacgctt
     1321 ctcgctgctc tttgagcctg cagacacctg gggggatacg gggaaaaggc ctccacggcc
     1381 actagtcttt cgtcttcaag aattcctcga gtttactccc tatcagtgat agagaacgta
     1441 tgaagagttt actccctatc agtgatagag aacgtatgca gactttactc cctatcagtg
     1501 atagagaacg tataaggagt ttactcccta tcagtgatag agaacgtatg accagtttac
     1561 tccctatcag tgatagagaa cgtatctaca gtttactccc tatcagtgat agagaacgta
     1621 tatccagttt actccctatc agtgatagag aacgtataag ctttaggcgt gtacggtggg
     1681 cgcctataaa agcagagctc gtttagtgaa ccgtcagatc gcctggagca attccacaac
     1741 acttttgtct tataccaact ttccgtacca cttcctaccc tcgtaaaggt ctagagctag
     1801 cgaattcgaa gccaccaact ggcaggtgcg tatgtcgagg taggcgtgta cggtgggagg
     1861 cctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagaatt ggctagcatc
     1921 aacaagtttg tacaaaaaag ttggcatgga gacagacaca ctcctgctat gggtactgct
     1981 gctctgggtt ccaggttcca ctggtgacgg aagccatcac caccatcatc acggtgcatc
     2041 tcctgaagat gaaattcagg cgttggaaca ggaaaacgcc caattggaac agaagattgc
     2101 agcgttgaag caagagaacg ctcagttaga acaagagatc caagctctgg agcaggacga
     2161 acaaaaactc atctcagaag aggatctgaa tgctgtgggc caggacacgc aggaggtcat
     2221 cgtggtgcca cactccttgc cctttaaggt ggtggtgatc tcagccatcc tggccctggt
     2281 ggtgctcacc atcatctccc ttatcatcct catcatgctt tggcagaaga agccacgtta
     2341 acggtacccc tcgactgtgc cttctagttg ccagacatct gttgtttgcc cctcccccgt
     2401 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2461 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2521 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctcttcgaa
     2581 tttaaatcgg atccagctga atctaagtgg atctgcgatc gctccggtgc ccgtcagtgg
     2641 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg
     2701 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     2761 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     2821 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
     2881 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     2941 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     3001 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     3061 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     3121 cgttacagat ctaagctgtg accggcgcct actctagagc cgccatgtct agactggaca
     3181 agagcaaagt cataaactct gctctggaat tactcaatgg agtcggtatc gaaggcctga
     3241 cgacaaggaa actcgctcaa aagctgggag ttgagcagcc taccctgtac tggcacgtga
     3301 agaacaagcg ggccctgctc gatgccctgc caatcgagat gctggacagg catcataccc
     3361 actcctgccc cctggaaggc gagtcatggc aagactttct gcggaacaac gccaagtcat
     3421 accgctgtgc tctcctctca catcgcgacg gggctaaagt gcatctcggc acccgcccaa
     3481 cagagaaaca gtacgaaacc ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct
     3541 ccctggagaa cgcactgtac gctctgtccg ccgtgggcca ctttacactg ggctgcgtat
     3601 tggaggaaca ggagcatcaa gtagcaaaag aggaaagaga gacacctacc accgattcta
     3661 tgcccccact tctgaaacaa gcaattgagc tgttcgaccg gcagggagcc gaacctgcct
     3721 tccttttcgg cctggaacta atcatatgtg gcctggagaa acagctaaag tgcgaaagcg
     3781 gcgggccgac cgacgccctt gacgattttg acttagacat gctcccagcc gatgcccttg
     3841 acgactttga ccttgatatg ctgcctgctg acgctcttga cgattttgac cttgacatgc
     3901 tccccgggga gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc
     3961 ctatggctag cgtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     4021 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     4081 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     4141 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     4201 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     4261 gggagcgcgt gatgaacttc gaggacggcg gcatggtgac cgtgacccag gactcctccc
     4321 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     4381 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     4441 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     4501 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     4561 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     4621 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaaggagg
     4681 gcagaggaag tcttctaaca tgcggtgacg tggaggagaa tcccggccct atgaccgagt
     4741 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4801 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4861 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4921 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4981 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     5041 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     5101 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     5161 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     5221 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     5281 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga aatcaacctc
     5341 tggattacaa aatttgtggg taaacttgtt tattgcagct tataatggtt acaaataaag
     5401 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5461 gtccaaactc atcaatgtat cttatcatgt ctggaattga ctcaaatgat gtcaattagt
     5521 ctatcagaag ctcatctggt ctcccttccg ggggacaaga catccctgtt taatatttaa
     5581 acagcagtgt tcccaaactg ggttcttata tcccttgctc tggtcaacca ggttgcaggg
     5641 tttcctgtcc tcacaggaac gaagtcccta aagaaacagt ggcagccagg tttagccccg
     5701 gaattgactg gattcctttt ttagggccca ttggtatggc tttttccccg tatcccccca
     5761 ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc
     5821 ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg
     5881 accggagcgg agccccgggc ggctcgctgc tgccccctag cgggggaggg acgtaattac
     5941 atccctgggg gctttggggg ggggctgtcc ctgatatcta taacaagaaa atatatatat
     6001 aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat gagttaaatc
     6061 ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg ttatagttca
     6121 aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa gcggcgactg
     6181 agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag caatatttca
     6241 agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaatctag ctgcatcagg
     6301 atcatatcgt cgggtctttt ttccggctca gtcatcgccc aagctggcgc tatctgggca
     6361 tcggggagga agaagcccgt gccttttccc gcgaggttga agcggcatgg aaagagtttg
     6421 ccgaggatga ctgctgctgc attgacgttg agcgaaaacg cacgtttacc atgatgattc
     6481 gggaaggtgt ggccatgcac gcctttaacg gtgaactgtt cgttcaggcc acctgggata
     6541 ccagttcgtc gcggcttttc cggacacagt tccggatggt cagcccgaag cgcatcagca
     6601 acccgaacaa taccggcgac agccggaact gccgtgccgg tgtgcagatt aatgacagcg
     6661 gtgcggcgct gggatattac gtcagcgagg acgggtatcc tggctggatg ccgcagaaat
     6721 ggacatggat accccgtgag ttacccggcg ggcgcgcttg gcgtaatcat ggtcatagct
     6781 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6841 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6901 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     6961 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7021 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7081 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7261 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     7621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7681 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7981 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8041 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8341 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8761 aataagggc
//
