LOCUS       sequence-366925-        9123 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Plasmid for inducible mammalian cell expression of the P3 helixCAM 
            and constitutive expression of an identifying fluorescent protein 
            (iRFP670). Flanked with PiggyBac sites for easy line construction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9123)
  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 9123)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_P3-PDGFR_EF1a_iRFP670
FEATURES             Location/Qualifiers
     source          1..9123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(115..219)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(245..700)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(382..403)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     594..613
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     827..849
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     841..858
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     842..858
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   988..1050
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1499..1877
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1507..1525
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1543..1561
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1579..1597
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1615..1633
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1651..1669
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1687..1705
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1723..1741
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1929..1967
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    2012..2036
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     2036..2098
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     CDS             2099..2125
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2234..2263
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2267..2413
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(2416..2440)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2445..2486
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2500..2517
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2574..2591)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2580..2787
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2864..3075
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3088..3356
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3390..4133
                     /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             4134..4187
                     /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             4188..5120
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     CDS             5121..5174
                     /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             5175..5774
                     /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(5175..5194)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5671..5691
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5807..5928
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5844..5863)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5898..5917
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   6691..6725
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7216..7232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7216..7232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7229..7251)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7240..7256
                     /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(7264..7270,7271..7288,7289..7294))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7309..7330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7447..7464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7618..8206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7698..7717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8377..9123,1..45,46..114))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALWPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9000..9019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
       61 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
      121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
      181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
      241 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
      301 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
      361 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
      421 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
      481 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
      541 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
      601 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
      661 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
      721 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
      781 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
      841 tgtaaaacga cggccagtga gcgcgcctcg ttcattcacg tttttgaacc cgtggaggac
      901 gggcagactc gcggtgcaaa tgtgttttac agcgtgatgg agcagatgaa gatgctcgac
      961 acgctgcaga acacgcagct agattaaccc tagaaagata atcatattgt gacgtacgtt
     1021 aaagataatc atgtgtaaaa ttgacgcatg tgttttatcg gtctgtatat cgaggtttat
     1081 ttattaattt gaatagatat taagttttat tatatttaca cttacatact aataataaat
     1141 tcaacaaaca atttatttat gtttatttat ttattaaaaa aaacaaaaac tcaaaatttc
     1201 ttctataaag taacaaaact tttatgaggg acagcccccc cccaaagccc ccagggatgt
     1261 aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc
     1321 ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg
     1381 tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg
     1441 gggggatacg gggaaaaggc ctccacggcc actagtcttt cgtcttcaag aattcctcga
     1501 gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc agtgatagag
     1561 aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt ttactcccta
     1621 tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa cgtatctaca
     1681 gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc agtgatagag
     1741 aacgtataag ctttaggcgt gtacggtggg cgcctataaa agcagagctc gtttagtgaa
     1801 ccgtcagatc gcctggagca attccacaac acttttgtct tataccaact ttccgtacca
     1861 cttcctaccc tcgtaaaggt ctagagctag cgaattcgaa gccaccaact ggcaggtgcg
     1921 tatgtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc
     1981 gtcagatcgc ctggagaatt ggctagcatc aacaagtttg tacaaaaaag ttggcatgga
     2041 gacagacaca ctcctgctat gggtactgct gctctgggtt ccaggttcca ctggtgacta
     2101 tccatatgat gttccagatt atgctggggc cagcccagag gatgaaatcc aacaactgga
     2161 ggaagagata gcgcaactgg aacagaaaaa cgcggctctc aaggagaaaa accaggccct
     2221 caaatacggg gacgaacaaa aactcatctc agaagaggat ctgaatgctg tgggccagga
     2281 cacgcaggag gtcatcgtgg tgccacactc cttgcccttt aaggtggtgg tgatctcagc
     2341 catcctggcc ctggtggtgc tcaccatcat ctcccttatc atcctcatca tgctttggca
     2401 gaagaagcca cgttaaccca actttcttgt acaaagtggt tgcaggaaag ccaatcccaa
     2461 acccactcct aggtctcgac agtacagcat agaccggtcc accaccacca ccaccactaa
     2521 ggatccgggg ttggggttgc gccttttcca agcacctgct acatagcggt acccctcgac
     2581 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2641 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2701 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2761 ggaagacaat agcaggcatg ctggggatgc ggtgggctct tcgaatttaa atcggatcca
     2821 gctgaatcta agtggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     2881 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     2941 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     3001 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     3061 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     3121 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     3181 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     3241 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     3301 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatctaag
     3361 ctgtgaccgg cgcctactct agagccgcca tgtctagact ggacaagagc aaagtcataa
     3421 actctgctct ggaattactc aatggagtcg gtatcgaagg cctgacgaca aggaaactcg
     3481 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     3541 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccactcc tgccccctgg
     3601 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcataccgc tgtgctctcc
     3661 tctcacatcg cgacggggct aaagtgcatc tcggcacccg cccaacagag aaacagtacg
     3721 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     3781 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     3841 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     3901 aacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     3961 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccgaccgacg
     4021 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     4081 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc ggggagggca
     4141 gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg gcgcgtaagg
     4201 tcgatctcac ctcctgcgat cgcgagccga tccacatccc cggcagcatt cagccgtgcg
     4261 gctgcctgct agcctgcgac gcgcaggcgg tgcggatcac gcgcattacg gaaaatgccg
     4321 gcgcgttctt tggacgcgaa actccgcggg tcggtgagct actcgccgat tacttcggcg
     4381 agaccgaagc ccatgcgctg cgcaacgcac tggcgcagtc ctccgatcca aagcgaccgg
     4441 cgctgatctt cggttggcgc gacggcctga ccggccgcac cttcgacatc tcactgcatc
     4501 gccatgacgg tacatcgatc atcgagttcg agcctgcggc ggccgaacag gccgacaatc
     4561 cgctgcggct gacgcggcag atcatcgcgc gcaccaaaga actgaagtcg ctcgaagaga
     4621 tggccgcacg ggtgccgcgc tatctgcagg cgatgctcgg ctatcaccgc gtgatgttgt
     4681 accgcttcgc ggacgacggc tccgggatgg tgatcggcga ggcgaagcgc agcgacctcg
     4741 agagctttct cggtcagcac tttccggcgt cgctggtccc gcagcaggcg cggctactgt
     4801 acttgaagaa cgcgatccgc gtggtctcgg attcgcgcgg catcagcagc cggatcgtgc
     4861 ccgagcacga cgcctccggc gccgcgctcg atctgtcgtt cgcgcacctg cgcagcatct
     4921 cgccctgcca tctcgaattt ctgcggaaca tgggcgtcag cgcctcgatg tcgctgtcga
     4981 tcatcattga cggcacgcta tggggattga tcatctgtca tcattacgag ccgcgtgccg
     5041 tgccgatggc gcagcgcgtc gcggccgaaa tgttcgccga cttcttatcg ctgcacttca
     5101 ccgccgccca ccaccaacgc gagggcagag gaagtcttct aacatgcggt gacgtggagg
     5161 agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     5221 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     5281 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     5341 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     5401 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     5461 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     5521 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     5581 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     5641 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     5701 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5761 agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgggtaaact tgtttattgc
     5821 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5881 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggaa
     5941 ttgactcaaa tgatgtcaat tagtctatca gaagctcatc tggtctccct tccgggggac
     6001 aagacatccc tgtttaatat ttaaacagca gtgttcccaa actgggttct tatatccctt
     6061 gctctggtca accaggttgc agggtttcct gtcctcacag gaacgaagtc cctaaagaaa
     6121 cagtggcagc caggtttagc cccggaattg actggattcc ttttttaggg cccattggta
     6181 tggctttttc cccgtatccc cccaggtgtc tgcaggctca aagagcagcg agaagcgttc
     6241 agaggaaagc gatcccgtgc caccttcccc gtgcccgggc tgtccccgca cgctgccggc
     6301 tcggggatgc ggggggagcg ccggaccgga gcggagcccc gggcggctcg ctgctgcccc
     6361 ctagcggggg agggacgtaa ttacatccct gggggctttg ggggggggct gtccctgata
     6421 tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa
     6481 tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt
     6541 gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct
     6601 cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat
     6661 ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc
     6721 tagggttaat ctagctgcat caggatcata tcgtcgggtc ttttttccgg ctcagtcatc
     6781 gccaaagctg gcgctatctg ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg
     6841 ttgaagcggc atggaaagag tttgccgagg atgactgctg ctgcattgac gttgagcgaa
     6901 aacgcacgtt taccatgatg attctggaag gtgtggccat gcacgccttt aacggtgaac
     6961 tgttcgttca ggccacctgg gataccagtt cgtcgcggct tttccggaca cagttccgga
     7021 tggtcagccc gaagcgcatc agcaacccga acaataccgg cgacagccgg aactgccgtg
     7081 ccggtgtgca gattaatgac agcggtgcgg cgctgggata ttacgtcagc gaggacgggt
     7141 atcctggctg gatgccgcag aaatggacat ggataccccg tgagttaccc ggcgggcgcg
     7201 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7261 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7321 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7381 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7441 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7501 tcactcaaat gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7561 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7621 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7681 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7741 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7801 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7861 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7921 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     7981 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8041 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8101 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8161 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8221 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8281 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8341 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8401 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactac ccgtcgtgta
     8461 gataactacg atacgggagg gcttaccatc tggccacagt gctgcaatga taccgcgaga
     8521 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8581 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8641 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8701 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8761 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8821 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8881 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     8941 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9001 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9061 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9121 acc
//
