LOCUS       sequence-346217-        9231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasma membrane localized FRB, rapamycin heterodimerization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9231)
  AUTHORS   Mahlandt EK, Palacios Martinez S, Arts JJG, Tol S, van Buul JD, 
            Goedhart J
  TITLE     Opto-RhoGEFs, an optimized optogenetic toolbox to reversibly control
            Rho GTPase activity on a global to subcellular scale, enabling 
            precise control over vascular endothelial barrier strength.
  JOURNAL   Elife. 2023 Jul 14;12:RP84364. doi: 10.7554/eLife.84364.
  PUBMED    37449837
REFERENCE   2  (bases 1 to 9231)
  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: "Elife."; 
            date: "2023-07-14"; volume: "12:RP84364. doi"; pages: " 
            10.7554/eLife.84364"
COMMENT     Sequence Label: pLV-Lck-FRB-mTurquoise2
FEATURES             Location/Qualifiers
     source          1..9231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        6..309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     464..484
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(594..611)
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
     CDS             626..904
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     CDS             911..937
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      944..953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(950..952,953..955,956..1663)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(995..1016)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1256..1275)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1603..1624
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1749..2301
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1893..1910)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2120..2139
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2328..2927
                     /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(2328..2347)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2824..2844
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3819..4052
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4124..4245
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4161..4180)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4215..4234
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4285..4420
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4440..4459)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4441..4459)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4469..4486)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4469..4485)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4478..4500)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4627..5082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4714..4733)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4924..4945
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5108..5212
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5213..5281,5282..6073)
                     /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     complement(5431..5450)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6244..6832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6733..6752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6986..7003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7120..7141
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7156..7161,7162..7179,7180..7186)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7194..7210
                     /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)."
     primer_bind     7199..7221
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7218..7234
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7218..7234
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7253..7273
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7255..7273
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7301..7527
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             7528..7708
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7755..7880
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8373..8606
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8791..8835
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    9097..9214
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
       61 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      121 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      181 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      481 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      541 caccatgggc tgcgtgtgca gcagcaaccc cgagagatct gcgaattcag gtgctggtgc
      601 tggtgctggt gctatcctat ctagaatcct ctggcatgag atgtggcatg aaggcctgga
      661 agaggcatct cgtttgtact ttggggaaag gaacgtgaaa ggcatgtttg aggtgctgga
      721 gcccttgcat gctatgatgg aacggggccc ccagactctg aaggaaacat cctttaatca
      781 ggcctatggt cgagatttaa tggaggccca agagtggtgc aggaagtaca tgaaatcagg
      841 gaatgtcaag gacctcctgc aagcctggga cctctattat catgtgttcc gacgaatctc
      901 aaagactagt tatccgtacg acgtaccaga ctacgcaccg gtcgccacca tggtgagcaa
      961 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1021 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1081 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1141 cctgtcctgg ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     1201 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1261 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     1321 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     1381 caactacttt agcgacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1441 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca
     1501 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     1561 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1621 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacagcatct tgagtccctt
     1681 tttaccgctg ttaccaattt tcttttgtct ttgggtaaac ccgggaattc cgcccctctc
     1741 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     1801 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     1861 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     1921 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     1981 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     2041 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     2101 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     2161 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     2221 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     2281 ttgaaaaaca cgatgataat atggatatgg ccacaacggt cgccaccatg accgagtaca
     2341 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     2401 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     2461 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     2521 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     2581 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     2641 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     2701 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     2761 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     2821 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     2881 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     2941 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atgtattcaa tctaagcagg
     3001 ctttcacttt ctcgccaact tacaaggcct ttctgtgtaa acaatacctg aacctttacc
     3061 ccgttgcccg gcaacggcca ggtctgtgcc aagtgtttgc tgacgcaacc cccactggct
     3121 ggggcttggt catgggccat cagcgcatgc gtggaacctt ttcggctcct ctgccgatcc
     3181 atactgcgga actcctagcc gcttgttttg ctcgcagcag gtctggagca aacattatcg
     3241 ggactgataa ctctgttgtc ctatcccgca aatatacatc gtttccatgg ctgctaggct
     3301 gtgctgccaa ctggatcctg cgcgggacgt cctttgttta cgtcccgtcg gcgctgaatc
     3361 ctgcggacga cccttctcgg ggtcgcttgg gactctctcg tccccttctc cgtctgccgt
     3421 tccgaccgac cacggggcgc acctctcttt acgcggactc cccgtctgtg ccttctcatc
     3481 tgccggaccg tgtgcacttc gcttcacctc tgcacgtcgc atggagacca ccgtgaacgc
     3541 ccaccaaata ttgcccaagg tcttacataa gaggactctt ggactctcag caatgtcaac
     3601 gaccgacctt gaggcatact tcaaagactg tttgtttaaa gactgggagg agttggggga
     3661 ggagattagg ttaaaggtct ttgtactagg aggctgtagg cataaattgg tctgcgcacc
     3721 agcaccatgg cgcaatcact agagcggggt acctttaaga ccaatgactt acaaggcagc
     3781 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     3841 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
     3901 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     3961 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     4021 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     4081 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc
     4141 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4201 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta
     4261 gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4321 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4381 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc
     4441 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     4501 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     4561 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     4621 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4681 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4741 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4801 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4861 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4921 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4981 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5041 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     5101 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5161 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5221 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5281 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5341 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5401 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5461 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5521 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5581 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5641 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5701 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5761 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5821 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5881 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5941 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6001 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6061 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6121 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6181 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6241 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6301 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6361 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     6421 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6481 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6541 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6601 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6661 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6721 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6781 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     6841 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     6901 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     6961 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     7021 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     7081 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     7141 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     7201 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     7261 ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc
     7321 ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag
     7381 caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa
     7441 cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta
     7501 tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg
     7561 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7621 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7681 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     7741 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     7801 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     7861 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     7921 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     7981 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     8041 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     8101 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     8161 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     8221 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     8281 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     8341 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     8401 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     8461 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     8521 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     8581 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     8641 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     8701 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     8761 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     8821 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     8881 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     8941 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     9001 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     9061 tccattcgat tagtgaacgg atctcgacgg tatcggtttt aaaagaaaag gggggattgg
     9121 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     9181 attacaaaaa caaattacaa aaattcaaaa ttttatcgat aagcttggga g
//
