LOCUS       sequence-337544-       10545 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of a transferrin receptor-SNAP-tag fusion in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10545)
  AUTHORS   FitzGerald LI, Aurelio L, Chen M, Yuen D, Rennick JJ, Graham B, 
            Johnston APR
  TITLE     A molecular sensor to quantify the localization of proteins, DNA and
            nanoparticles in cells.
  JOURNAL   Nat Commun. 2020 Sep 8;11(1):4482. doi: 10.1038/s41467-020-18082-8.
  PUBMED    32901011
REFERENCE   2  (bases 1 to 10545)
  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: "Nat 
            Commun."; date: "2020-09-8"; volume: "11(1):4482. doi"; pages: " 
            10.1038/s41467-020-18082-8"
COMMENT     Sequence Label: pCDH-EF1-TfR20-SNAP-IRES-Puro
FEATURES             Location/Qualifiers
     source          1..10545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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    1807..1923
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1966..2177
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2190..2458
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      4855..4864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4861..5406
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     misc_feature    join(5419..5993,5994..5996,5997..6005)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5586..5603)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5813..5832
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6018..6617
                     /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(6018..6037)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6514..6534
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6624..7212
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6677..6697)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7095..7106)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7286..7519
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7591..7712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7628..7647)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7682..7701
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7752..7887
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7920..7936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7920..7936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7933..7955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7944..7960
                     /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(7968..7974,7975..7992,7993..7998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8013..8034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8151..8168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8322..8910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(9081..9872,9873..9941))
                     /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     9704..9723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9942..10046)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10114..10132
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(10170..10192)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     10292..10311
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     10505..10527
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10519..10536
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10520..10536
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat
     1921 tttatcgata ctagtggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     1981 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     2041 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     2101 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     2161 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     2221 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     2281 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     2341 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     2401 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     2461 agctgtgacc ggcgcctact ctagagctag cgctaccgga ctcagatctc gagcgccacc
     2521 atgatggatc aagctagatc agcattctct aacttgtttg gtggagaacc attgtcatat
     2581 acccggttca gcctggctcg gcaagtagat ggcgataaca gtcatgtgga gatgaaactt
     2641 gctgtagatg aagaagaaaa tgctgacaat aacacaaagg ccaatgtcac aaaaccaaaa
     2701 aggtgtagtg gaagtatctg ctatgggact attgctgtga tcgtcttttt cttgattgga
     2761 tttatgattg gctacttggg ctattgtaaa ggggtagaac caaaaactga gtgtgagaga
     2821 ctggcaggaa ccgagtctcc agtgagggag gagccaggag aggacttccc tgcagcacgt
     2881 cgcttatatt gggatgacct gaagagaaag ttgtcggaga aactggacag cacagacttc
     2941 accagcacca tcaagctgct gaatgaaaat tcatatgtcc ctcgtgaggc tggatctcaa
     3001 aaagatgaaa atcttgcgtt gtatgttgaa aatcaatttc gtgaatttaa actcagcaaa
     3061 gtctggcgtg atcaacattt tgttaagatt caggtcaaag acagcgctca aaactcggtg
     3121 atcatagttg ataagaacgg tagacttgtt tacctggtgg agaatcctgg gggttatgtg
     3181 gcgtatagta aggctgcaac agttactggt aaactggtcc atgctaattt tggtactaaa
     3241 aaagattttg aggatttata cactcctgtg aatggatcta tagtgattgt cagagcaggg
     3301 aaaatcacct ttgcagaaaa ggttgcaaat gctgaaagct taaatgcaat tggtgtgttg
     3361 atatacatgg accagactaa atttcccatt gttaacgcag aactttcatt ctttggacat
     3421 gctcatctgg ggacaggtga cccttacaca cctggattcc cttccttcaa ccacactcag
     3481 tttccaccat ctcggtcatc aggattgcct aatatacctg tccagacaat ctccagagct
     3541 gctgcagaaa agctgtttgg gaatatggaa ggagactgtc cctctgactg gaaaacagac
     3601 tctacatgta ggatggtaac ctcagaaagc aagaatgtga agctcactgt gagcaatgtg
     3661 ctgaaagaga taaaaattct taacatcttt ggagttatta aaggctttgt agaaccagat
     3721 cactatgttg tagttggggc ccagagagat gcatggggcc ctggagctgc aaaatccggt
     3781 gtaggcacag ctctcctatt gaaacttgcc cagatgttct cagatatggt cttaaaagat
     3841 gggtttcagc ccagcagaag cattatcttt gccagttgga gtgctggaga ctttggatcg
     3901 gttggtgcca ctgaatggct agagggatac ctttcgtccc tgcatttaaa ggctttcact
     3961 tatattaatc tggataaagc ggttcttggt accagcaact tcaaggtttc tgccagccca
     4021 ctgttgtata cgcttattga gaaaacaatg caaaatgtga agcatccggt tactgggcaa
     4081 tttctatatc aggacagcaa ctgggccagc aaagttgaga aactcacttt agacaatgct
     4141 gctttccctt tccttgcata ttctggaatc ccagcagttt ctttctgttt ttgcgaggac
     4201 acagattatc cttatttggg taccaccatg gacacctata aggaactgat tgagaggatt
     4261 cctgagttga acaaagtggc acgagcagct gcagaggtcg ctggtcagtt cgtgattaaa
     4321 ctaacccatg atgttgaatt gaacctggac tatgagaggt acaacagcca actgctttca
     4381 tttgtgaggg atctgaacca atacagagca gacataaagg aaatgggcct gagtttacag
     4441 tggctgtatt ctgctcgtgg agacttcttc cgtgctactt ccagactaac aacagatttc
     4501 gggaatgctg agaaaacaga cagatttgtc atgaagaaac tcaatgatcg tgtcatgaga
     4561 gtggagtatc acttcctctc tccctacgta tctccaaaag agtctccttt ccgacatgtc
     4621 ttctggggct ccggctctca cacgctgcca gctttactgg agaacttgaa actgcgtaaa
     4681 caaaataacg gtgcttttaa tgaaacgctg ttcagaaacc agttggctct agctacttgg
     4741 actattcagg gagctgcaaa tgccctctct ggtgacgttt gggacattga caatgagttt
     4801 tcagaattcg gctccaccgg ctccaccggc tccaccggcg cggatccacc ggtcgccacc
     4861 atggacaaag actgcgaaat gaagcgcacc accctggata gccctctggg caagctggaa
     4921 ctgtctgggt gcgaacaggg cctgcaccgt atcatcttcc tgggcaaagg aacatctgcc
     4981 gccgacgccg tggaagtgcc tgccccagcc gccgtgctgg gcggaccaga gccactgatg
     5041 caggccaccg cctggctcaa cgcctacttt caccagcctg aggccatcga ggagttccct
     5101 gtgccagccc tgcaccaccc agtgttccag caggagagct ttacccgcca ggtgctgtgg
     5161 aaactgctga aagtggtgaa gttcggagag gtcatcagct acagccacct ggccgccctg
     5221 gccggcaatc ccgccgccac cgccgccgtg aaaaccgccc tgagcggaaa tcccgtgccc
     5281 attctgatcc cctgccaccg ggtggtgcag ggcgacctgg acgtgggggg ctacgagggc
     5341 gggctcgccg tgaaagagtg gctgctggcc cacgagggcc acagactggg caagcctggg
     5401 ctgggttaag cggccgcgcc cctctccctc ccccccccct aacgttactg gccgaagccg
     5461 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     5521 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     5581 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     5641 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     5701 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     5761 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     5821 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     5881 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     5941 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca
     6001 caaccatggc gtccggaatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     6061 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     6121 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     6181 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     6241 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     6301 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     6361 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     6421 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     6481 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     6541 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     6601 gcaagcccgg tgcctaggtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     6661 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     6721 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     6781 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     6841 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     6901 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     6961 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     7021 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     7081 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     7141 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     7201 ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     7261 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aagataagat
     7321 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7381 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     7441 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     7501 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     7561 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     7621 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     7681 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     7741 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     7801 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     7861 agtagtgagg aggctttttt ggaggcctag acttttgcag agaccaaatt cgtaatcatg
     7921 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     7981 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     8041 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     8101 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     8161 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     8221 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     8281 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     8341 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     8401 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     8461 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     8521 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     8581 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     8641 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     8701 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     8761 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     8821 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     8881 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8941 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     9001 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     9061 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     9121 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     9181 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     9241 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     9301 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     9361 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     9421 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     9481 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     9541 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     9601 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     9661 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     9721 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     9781 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     9841 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     9901 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     9961 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
    10021 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
    10081 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
    10141 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
    10201 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
    10261 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
    10321 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
    10381 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
    10441 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
    10501 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctg
//
