LOCUS       sequence-365749-        8940 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible mammalian cell expression of the Z17 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 8940)
  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 8940)
  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_Z17-PDGFR_EF1a_mCherry
FEATURES             Location/Qualifiers
     source          1..8940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(97..201)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(227..682)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(364..385)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     576..595
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     809..831
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     823..840
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     824..840
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   970..1032
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1481..1859
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     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"
     protein_bind    1669..1687
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1705..1723
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1911..1949
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    1994..2018
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     2018..2077
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2081..2107
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2252..2281
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2285..2431
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(2434..2458)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2463..2504
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2518..2535
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2592..2609)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2598..2805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2882..3093
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3106..3374
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3408..4151
                     /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             4152..4205
                     /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             4215..4919
                     /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(4357..4375)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4625..4644
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4920..4973
                     /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             4974..5573
                     /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(4974..4993)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5470..5490
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5606..5727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5643..5662)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5697..5716
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6490..6524
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7015..7031)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7015..7031)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7028..7050)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7039..7055
                     /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(7063..7069,7070..7087,7088..7093))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7108..7129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7246..7263)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7417..8005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7497..7516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8176..8940,1..27,28..96))
                     /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     8799..8818
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
       61 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
      121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
      181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
      241 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
      301 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
      361 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
      421 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
      481 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
      541 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
      601 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
      661 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
      721 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
      781 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
      841 gagcgcgcct cgttcattca cgtttttgaa cccgtggagg acgggcagac tcgcggtgca
      901 aatgtgtttt acagcgtgat ggagcagatg aagatgctcg acacgctgca gaacacgcag
      961 ctagattaac cctagaaaga taatcatatt gtgacgtacg ttaaagataa tcatgtgtaa
     1021 aattgacgca tgtgttttat cggtctgtat atcgaggttt atttattaat ttgaatagat
     1081 attaagtttt attatattta cacttacata ctaataataa attcaacaaa caatttattt
     1141 atgtttattt atttattaaa aaaaacaaaa actcaaaatt tcttctataa agtaacaaaa
     1201 cttttatgag ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg
     1261 ctagggggca gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc
     1321 ccgagccggc agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc
     1381 ctctgaacgc ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaag
     1441 gcctccacgg ccactagtct ttcgtcttca agaattcctc gagtttactc cctatcagtg
     1501 atagagaacg tatgaagagt ttactcccta tcagtgatag agaacgtatg cagactttac
     1561 tccctatcag tgatagagaa cgtataagga gtttactccc tatcagtgat agagaacgta
     1621 tgaccagttt actccctatc agtgatagag aacgtatcta cagtttactc cctatcagtg
     1681 atagagaacg tatatccagt ttactcccta tcagtgatag agaacgtata agctttaggc
     1741 gtgtacggtg ggcgcctata aaagcagagc tcgtttagtg aaccgtcaga tcgcctggag
     1801 caattccaca acacttttgt cttataccaa ctttccgtac cacttcctac cctcgtaaag
     1861 gtctagagct agcgaattcg aagccaccaa ctggcaggtg cgtatgtcga ggtaggcgtg
     1921 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagaa
     1981 ttggctagca tcaacaagtt tgtacaaaaa agttggcatg gagacagaca cactcctgct
     2041 atgggtactg ctgctctggg ttccaggttc cactggtgac tatccatatg atgttccaga
     2101 ttatgctggg gccaatgaga aagaagaact gaagagtaag aaggctgagc tgcgaaacag
     2161 aattgaacaa ctgaagcaga agcgagagca actcaagcag aaaatagcta atctgcgcaa
     2221 ggaaatagaa gcatacaaac tgcaggtcga cgaacaaaaa ctcatctcag aagaggatct
     2281 gaatgctgtg ggccaggaca cgcaggaggt catcgtggtg ccacactcct tgccctttaa
     2341 ggtggtggtg atctcagcca tcctggccct ggtggtgctc accatcatct cccttatcat
     2401 cctcatcatg ctttggcaga agaagccacg ttaacccaac tttcttgtac aaagtggttg
     2461 caggaaagcc aatcccaaac ccactcctag gtctcgacag tacagcatag accggtccac
     2521 caccaccacc accactaagg atccggggtt ggggttgcgc cttttccaag cacctgctac
     2581 atagcggtac ccctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2641 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2701 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2761 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctcttc
     2821 gaatttaaat cggatccagc tgaatctaag tggatctgcg atcgctccgg tgcccgtcag
     2881 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     2941 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3001 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3061 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     3121 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     3181 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     3241 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     3301 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     3361 cgccgttaca gatctaagct gtgaccggcg cctactctag agccgccatg tctagactgg
     3421 acaagagcaa agtcataaac tctgctctgg aattactcaa tggagtcggt atcgaaggcc
     3481 tgacgacaag gaaactcgct caaaagctgg gagttgagca gcctaccctg tactggcacg
     3541 tgaagaacaa gcgggccctg ctcgatgccc tgccaatcga gatgctggac aggcatcata
     3601 cccactcctg ccccctggaa ggcgagtcat ggcaagactt tctgcggaac aacgccaagt
     3661 cataccgctg tgctctcctc tcacatcgcg acggggctaa agtgcatctc ggcacccgcc
     3721 caacagagaa acagtacgaa accctggaaa atcagctcgc gttcctgtgt cagcaaggct
     3781 tctccctgga gaacgcactg tacgctctgt ccgccgtggg ccactttaca ctgggctgcg
     3841 tattggagga acaggagcat caagtagcaa aagaggaaag agagacacct accaccgatt
     3901 ctatgccccc acttctgaaa caagcaattg agctgttcga ccggcaggga gccgaacctg
     3961 ccttcctttt cggcctggaa ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa
     4021 gcggcgggcc gaccgacgcc cttgacgatt ttgacttaga catgctccca gccgatgccc
     4081 ttgacgactt tgaccttgat atgctgcctg ctgacgctct tgacgatttt gaccttgaca
     4141 tgctccccgg ggagggcaga ggaagtcttc taacatgcgg tgacgtggag gagaatcccg
     4201 gccctatggc tagcgtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     4261 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     4321 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     4381 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     4441 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     4501 agtgggagcg cgtgatgaac ttcgaggacg gcggcatggt gaccgtgacc caggactcct
     4561 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     4621 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     4681 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     4741 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     4801 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     4861 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagg
     4921 agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc cctatgaccg
     4981 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     5041 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     5101 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     5161 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     5221 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     5281 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     5341 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5401 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5461 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5521 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac
     5581 ctctggatta caaaatttgt gggtaaactt gtttattgca gcttataatg gttacaaata
     5641 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5701 tttgtccaaa ctcatcaatg tatcttatca tgtctggaat tgactcaaat gatgtcaatt
     5761 agtctatcag aagctcatct ggtctccctt ccgggggaca agacatccct gtttaatatt
     5821 taaacagcag tgttcccaaa ctgggttctt atatcccttg ctctggtcaa ccaggttgca
     5881 gggtttcctg tcctcacagg aacgaagtcc ctaaagaaac agtggcagcc aggtttagcc
     5941 ccggaattga ctggattcct tttttagggc ccattggtat ggctttttcc ccgtatcccc
     6001 ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc
     6061 accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc
     6121 cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat
     6181 tacatccctg ggggctttgg gggggggctg tccctgatat ctataacaag aaaatatata
     6241 tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa
     6301 atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt
     6361 tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga
     6421 ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt
     6481 tcaagaatgc atgcgtcaat tttacgcaga ctatctttct agggttaatc tagctgcatc
     6541 aggatcatat cgtcgggtct tttttccggc tcagtcatcg cccaagctgg cgctatctgg
     6601 gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt
     6661 ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa acgcacgttt accatgatga
     6721 ttcgggaagg tgtggccatg cacgccttta acggtgaact gttcgttcag gccacctggg
     6781 ataccagttc gtcgcggctt ttccggacac agttccggat ggtcagcccg aagcgcatca
     6841 gcaacccgaa caataccggc gacagccgga actgccgtgc cggtgtgcag attaatgaca
     6901 gcggtgcggc gctgggatat tacgtcagcg aggacgggta tcctggctgg atgccgcaga
     6961 aatggacatg gataccccgt gagttacccg gcgggcgcgc ttggcgtaat catggtcata
     7021 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7081 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7141 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7201 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7261 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7321 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7381 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7441 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7501 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7561 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7621 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7681 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7741 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7801 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     7861 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7921 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7981 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8101 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8161 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8221 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8281 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8341 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8401 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8461 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8521 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8581 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8641 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8701 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8761 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8821 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8881 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
//
