LOCUS       sequence-366942-        8868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible mammalian cell expression of the P9 helixCAM 
            and constitutive expression of an identifying fluorescent protein 
            (mPlum). Flanked with PiggyBac sites for easy line construction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8868)
  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 8868)
  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_P9-PDGFR_EF1a_mPlum
FEATURES             Location/Qualifiers
     source          1..8868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(92..196)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(222..677)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(359..380)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     571..590
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     804..826
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     818..835
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     819..835
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   965..1027
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1476..1854
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1484..1502
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1520..1538
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1556..1574
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1592..1610
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1628..1646
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1664..1682
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1700..1718
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1906..1944
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    1989..2013
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     2013..2072
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2076..2102
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2211..2240
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2244..2390
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(2393..2417)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2422..2463
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2477..2494
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2551..2568)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2557..2764
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2841..3052
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3065..3333
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3367..4110
                     /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             4111..4164
                     /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             4165..4842
                     /codon_start=1
                     /product="far-red monomeric derivative of DsRed fluorescent
                     protein (Wang et al., 2004)"
                     /label=mPlum
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK
                     MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGA"
     primer_bind     4566..4585
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4760..4783
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             4843..4896
                     /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             4897..5496
                     /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(4897..4916)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5393..5413
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5529..5650
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5566..5585)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5620..5639
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6413..6447
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6938..6954)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6938..6954)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6951..6973)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6962..6978
                     /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(6986..6992,6993..7010,7011..7016))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7031..7052
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7169..7186)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7340..7928)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7420..7439)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8099..8868,1..22,23..91))
                     /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     8722..8741
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
       61 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
      121 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
      181 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
      241 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
      301 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
      361 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
      421 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
      481 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
      541 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
      601 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
      661 gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc
      721 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      781 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg
      841 cgcctcgttc attcacgttt ttgaacccgt ggaggacggg cagactcgcg gtgcaaatgt
      901 gttttacagc gtgatggagc agatgaagat gctcgacacg ctgcagaaca cgcagctaga
      961 ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg tgtaaaattg
     1021 acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa tagatattaa
     1081 gttttattat atttacactt acatactaat aataaattca acaaacaatt tatttatgtt
     1141 tatttattta ttaaaaaaaa caaaaactca aaatttcttc tataaagtaa caaaactttt
     1201 atgagggaca gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg
     1261 gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag
     1321 ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg
     1381 aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaggcctc
     1441 cacggccact agtctttcgt cttcaagaat tcctcgagtt tactccctat cagtgataga
     1501 gaacgtatga agagtttact ccctatcagt gatagagaac gtatgcagac tttactccct
     1561 atcagtgata gagaacgtat aaggagttta ctccctatca gtgatagaga acgtatgacc
     1621 agtttactcc ctatcagtga tagagaacgt atctacagtt tactccctat cagtgataga
     1681 gaacgtatat ccagtttact ccctatcagt gatagagaac gtataagctt taggcgtgta
     1741 cggtgggcgc ctataaaagc agagctcgtt tagtgaaccg tcagatcgcc tggagcaatt
     1801 ccacaacact tttgtcttat accaactttc cgtaccactt cctaccctcg taaaggtcta
     1861 gagctagcga attcgaagcc accaactggc aggtgcgtat gtcgaggtag gcgtgtacgg
     1921 tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagaattggc
     1981 tagcatcaac aagtttgtac aaaaaagttg gcatggagac agacacactc ctgctatggg
     2041 tactgctgct ctgggttcca ggttccactg gtgactatcc atatgatgtt ccagattatg
     2101 ctggggccag cccggaggat gaaaatcaag cccttgagca aaaaaatgct caattgaaac
     2161 aagagattgc ggcccttgaa caggagatcg cgcagcttga atacggtgac gaacaaaaac
     2221 tcatctcaga agaggatctg aatgctgtgg gccaggacac gcaggaggtc atcgtggtgc
     2281 cacactcctt gccctttaag gtggtggtga tctcagccat cctggccctg gtggtgctca
     2341 ccatcatctc ccttatcatc ctcatcatgc tttggcagaa gaagccacgt taacccaact
     2401 ttcttgtaca aagtggttgc aggaaagcca atcccaaacc cactcctagg tctcgacagt
     2461 acagcataga ccggtccacc accaccacca ccactaagga tccggggttg gggttgcgcc
     2521 ttttccaagc acctgctaca tagcggtacc cctcgactgt gccttctagt tgccagccat
     2581 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     2641 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     2701 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     2761 gggatgcggt gggctcttcg aatttaaatc ggatccagct gaatctaagt ggatctgcga
     2821 tcgctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     2881 gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag
     2941 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     3001 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agctgaagct
     3061 tcgaggggct cgcatctctc cttcacgcgc ccgccgccct acctgaggcc gccatccacg
     3121 ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc tcctgaactg cgtccgccgt
     3181 ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt ccggcgctcc cttggagcct
     3241 acctagactc agccggctct ccacgctttg cctgaccctg cttgctcaac tctacgtctt
     3301 tgtttcgttt tctgttctgc gccgttacag atctaagctg tgaccggcgc ctactctaga
     3361 gccgccatgt ctagactgga caagagcaaa gtcataaact ctgctctgga attactcaat
     3421 ggagtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag
     3481 cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag
     3541 atgctggaca ggcatcatac ccactcctgc cccctggaag gcgagtcatg gcaagacttt
     3601 ctgcggaaca acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa
     3661 gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg
     3721 ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc
     3781 cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga
     3841 gagacaccta ccaccgattc tatgccccca cttctgaaac aagcaattga gctgttcgac
     3901 cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag
     3961 aaacagctaa agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac
     4021 atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt
     4081 gacgattttg accttgacat gctccccggg gagggcagag gaagtcttct aacatgcggt
     4141 gacgtggagg agaatcccgg ccctatggtg agcaagggcg aggaggtcat caaggagttc
     4201 atgcgcttca aggagcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     4261 gagggcgagg gccgccccta cgagggcacc cagaccgcca ggctgaaggt gaccaagggt
     4321 ggccccctgc ccttcgcctg ggacatcctg tcccctcaga tcatgtacgg ctccaaggcc
     4381 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     4441 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     4501 tccctgcagg acggcgagtt catctacaag gtgaaggtgc gcggcaccaa cttcccctcc
     4561 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     4621 cccgaggacg gcgccctgaa gggcgagatg aagatgaggc tgaggctgaa ggacggcggc
     4681 cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca gctgcccggc
     4741 gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta caccatcgtg
     4801 gaacagtacg agcgcgccga gggccgccac tccaccggcg ccgagggcag aggaagtctt
     4861 ctaacatgcg gtgacgtgga ggagaatccc ggccctatga ccgagtacaa gcccacggtg
     4921 cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc
     4981 gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag
     5041 ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac
     5101 gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc
     5161 gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag
     5221 atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc
     5281 ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg
     5341 gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc
     5401 cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg
     5461 cgcacctggt gcatgacccg caagcccggt gcctgaaatc aacctctgga ttacaaaatt
     5521 tgtgggtaaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5581 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5641 atgtatctta tcatgtctgg aattgactca aatgatgtca attagtctat cagaagctca
     5701 tctggtctcc cttccggggg acaagacatc cctgtttaat atttaaacag cagtgttccc
     5761 aaactgggtt cttatatccc ttgctctggt caaccaggtt gcagggtttc ctgtcctcac
     5821 aggaacgaag tccctaaaga aacagtggca gccaggttta gccccggaat tgactggatt
     5881 ccttttttag ggcccattgg tatggctttt tccccgtatc cccccaggtg tctgcaggct
     5941 caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt gccaccttcc ccgtgcccgg
     6001 gctgtccccg cacgctgccg gctcggggat gcggggggag cgccggaccg gagcggagcc
     6061 ccgggcggct cgctgctgcc ccctagcggg ggagggacgt aattacatcc ctgggggctt
     6121 tggggggggg ctgtccctga tatctataac aagaaaatat atatataata agttatcacg
     6181 taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa aagtcacgta
     6241 aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat cagtgacact
     6301 taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat gtcctaaatg
     6361 cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa tgcatgcgtc
     6421 aattttacgc agactatctt tctagggtta atctagctgc atcaggatca tatcgtcggg
     6481 tcttttttcc ggctcagtca tcgcccaagc tggcgctatc tgggcatcgg ggaggaagaa
     6541 gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag agtttgccga ggatgactgc
     6601 tgctgcattg acgttgagcg aaaacgcacg tttaccatga tgattcggga aggtgtggcc
     6661 atgcacgcct ttaacggtga actgttcgtt caggccacct gggataccag ttcgtcgcgg
     6721 cttttccgga cacagttccg gatggtcagc ccgaagcgca tcagcaaccc gaacaatacc
     6781 ggcgacagcc ggaactgccg tgccggtgtg cagattaatg acagcggtgc ggcgctggga
     6841 tattacgtca gcgaggacgg gtatcctggc tggatgccgc agaaatggac atggataccc
     6901 cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     6961 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     7021 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     7081 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     7141 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     7201 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     7261 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     7321 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     7381 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     7441 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     7501 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     7561 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     7621 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     7681 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     7741 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     7801 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     7861 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     7921 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     7981 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     8041 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     8101 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     8161 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     8221 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     8281 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     8341 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     8401 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     8461 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     8521 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     8581 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     8641 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     8701 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     8761 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     8821 gttcgatgta acccactcgt gcacccaact gatcttcagc atctttta
//
