pLV-EF1a-FRB-mCherry-MinD-IRES-Puro

LOCUS       pLV-EF1a-FRB-mCherry-MinD-IRES-Puro               10526 bp ds-DNA     circular SYN 26-JAN-2024
DEFINITION  Expression of FRB-mCherry-MinD in mammalian cells for payload 
            delivery.
ACCESSION   .
VERSION     .
KEYWORDS    pLV-EF1a-FRB-mCherry-MinD-IRES-Puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10526)
  AUTHORS   Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM
  TITLE     A programmable reaction-diffusion system for spatiotemporal cell 
            signaling circuit design.
  JOURNAL   Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi: 
            10.1016/j.cell.2023.12.007.
  PUBMED    38181787
REFERENCE   2  (bases 1 to 10526)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 26, 2024 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10526
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(286..330)
                     /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    515..748
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1245..1370
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             1417..2050
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     polyA_signal    2218..2352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2242..2261)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2296..2315
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2400..2504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2505..3365
                     /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(2723..2742)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3536..4124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4025..4044
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4278..4295
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4412..4433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4448..4478
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4486..4502
                     /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     4491..4513
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4510..4526
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4510..4526
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     LTR             4654..5287
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    5494..6082
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5600..5611
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6009..6029
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6106..6705)
                     /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(6189..6209)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6686..6705
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(6867..6886)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     7096..7113
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(8204..8911)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(8479..8498)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     8748..8766
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(8912..9181)
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     regulatory      9181..9190
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     intron          9206..10144
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(9229..9249)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        complement(10145..10356)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    10434..23
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
ORIGIN
        1 ccaatccccc cttttctttt aaaggcgata ccgtcgagat ccgttcacta atcgaatgga
       61 tctgtctctg tctctctctc caccttcttc ttctattcct tcgggcctgt cgggtcccct
      121 cggggttggg aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac
      181 tatagaaagt acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa
      241 taattttata taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt
      301 atctaattcc aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa
      361 ggagtgtatt aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg
      421 attccaaatc tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt
      481 gcaaatgagt tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct
      541 ttccacagcc aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag
      601 atgctgttgc gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa
      661 taattgtctg gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc
      721 tcccaagaac ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac
      781 tcttctcttt gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt
      841 atataattca cttctccaat tgtccctcat atctcctcct ccaggtctga agatcagcgg
      901 ccggccgctt gctgtgcggt ggtcttactt ttgttttgct cttcctctat cttgtctaaa
      961 gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt tgctactgta
     1021 ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag ctgtcccagt
     1081 atttgtctac agccttctga tgtttctaac aggccaggat taactgcgaa tcgttctagc
     1141 tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc tggccttaac
     1201 cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct ctcgcaccca
     1261 tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag tcgccgcccc
     1321 tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt
     1381 ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc cacactgact
     1441 aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt
     1501 gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga
     1561 gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa agcagctgct
     1621 tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg cctccctgga
     1681 aagtccccag cggaaagtcc cttgtagcaa gctcgatatc agcagttctt gaagtactcc
     1741 ggatgcagct ctcgggccac gtgatgaaat gctaggcggc tgtcaaacct ccactctaac
     1801 acttctctct ccgggtcatc catcccatgc aggctcacag ggtgtaacaa gctggtgttc
     1861 tctcctttat tggcctcttc taccttatct ggctcaactg gtactagctt gtagcaccat
     1921 ccaaaggtca gtggatatct gacccctggc cctggtgtgt agttctgcta atcagggaag
     1981 tagccttgtg tgtggtagat ccacagatca aggatatctt gtcttctttg ggagtgaatt
     2041 agcccttcca actactaagt ttgtagtaca tatttaacaa atacaatttc tttaaaatga
     2101 aaataattca gaggaatcac aggtttagag taaatgaaac cacaggtaat tggcagtggt
     2161 aatagggtat ggggtgggaa gtttgggatg attttggtta gcttgagtta tccagttgat
     2221 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2281 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2341 aataaacaag ttgatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc
     2401 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2881 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     2941 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3661 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3901 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4021 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4081 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4141 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4201 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4261 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     4321 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     4381 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     4441 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     4501 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
     4561 gtaaacgcta gcaatgtcaa ggcctctcac tctctgatat tcatttcttt gcaagttata
     4621 aatactgaat aataagatga catgaactac tactgctaga gattttccac actgactaaa
     4681 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
     4741 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
     4801 cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc agctgcttat
     4861 atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct ccctggaaag
     4921 tccccagcgg aaagtccctt gtagcaagct cgatatcagc agttcttgaa gtactccgga
     4981 tgcagctctc gggccacgtg atgaaatgct aggcggctgt caaacctcca ctctaacact
     5041 tctctctccg ggtcatccat cccatgcagg ctcacagggt gtaacaagct ggtgttctct
     5101 cctttattgg cctcttctac cttatctggc tcaactggta ctagcttgta gcaccatcca
     5161 aaggtcagtg gatatctgac ccctggccct ggtgtgtagt tctgctaatc agggaagtag
     5221 ccttgtgtgt ggtagatcca cagatcaagg atatcttgtc ttcgttggga gtgaattagc
     5281 ccttccagtc ccctcttttc ttttaaaaag tggctaagat ctacagctgc cttgtaagtc
     5341 attggtctta aaggtacctg aggtgtgact ggaaaaccca cctcctcctc ctcttgtgct
     5401 tctagccagg cacaatcagc attggtagct gctgtattgc tacttgtgat tgctccatgt
     5461 ttttctaggt ctcgactgca gaattaattc caggcgggga ggcggcccaa agggagatcc
     5521 gactcgtctg agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc
     5581 gggaaggaag gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca
     5641 gaatccaggt ggcaacacag gcgagcagcc atggaaagga cgtcagcttc cccgacaaca
     5701 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
     5761 cgatgagttc cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac
     5821 aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc
     5881 cacgttgcct gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca
     5941 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
     6001 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
     6061 ttttgtaatc cagaggttga ttgttccaga cgcgccccgc ccgggtcagg caccgggctt
     6121 gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa
     6181 gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac
     6241 cccggcgcgc tcggccgcct ccactccggg gagcacgacg gcgctgccca gacccttgcc
     6301 ctggtggtcg ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg
     6361 cggcgccagg aggccttcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc
     6421 catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca
     6481 gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg
     6541 cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt
     6601 gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac
     6661 gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatggtat tatcgtgttt
     6721 ttcaaaggaa aaccacgtcc ccgtggttcg gggggcctag acgttttttt aacctcgact
     6781 aaacacatgt aaagcatgtg caccgaggcc ccagatcaga tcccatacaa tggggtacct
     6841 tctgggcatc cttcagcccc ttgttgaata cgcttgagga gagccatttg actctttcca
     6901 caactatcca actcacaacg tggcactggg gttgtgccgc ctttgcaggt gtatcttata
     6961 cacgtggctt ttggccgcag aggcacctgt cgccaggtgg ggggttccgc tgcctgcaaa
     7021 gggtcgctac agacgttgtt tgtcttcaag aagcttccag aggaactgct tccttcacga
     7081 cattcaacag accttgcatt cctttggcga gaggggaaag acccctagga atgctcgtca
     7141 agaagacagg gccaggtttc cgggccctca cattgccaaa agacggcaat atggtggaaa
     7201 ataacatata gacaaacgca caccggcctt attccaagcg gcttcggcca gtaacgttag
     7261 gggggggggg cggaattgat cccgctagag ggaaaccgtt gctagcttaa gttagttaac
     7321 aattgttacc gcggcccgtc gactctagag cggccgctta tcccccgaac agccgtttaa
     7381 ggaatccctt cttttcctcc tctatgaagc ggaagggccg ttcttccccg aggagccgtt
     7441 ctaccgtatc ggcataagct ttcccagcat ccgcatttat gtcaagtatg acaggctccc
     7501 cctgattaga tgcgcggagg acggactggt cttccgggat aacgcctacc agttttatcc
     7561 tcagaatctc gaggacatct tccattgaaa gcatatctcc ccgagatact cgaccggggt
     7621 tatatcgggt aagcaggaga tgctccttaa ttggctcctc cccgttttct gccctccgtg
     7681 atttgctggc gagtatcccc agtattcggt ctgaatctcg cacactagac acctccggat
     7741 tagttgtgat tatggcttca tctgcgaagt acagtgccat aagtgcccct gtctctatac
     7801 cggcgggaga gtcacaaact atgaactcaa agtccatcgc tttcaagtca tccaaaactt
     7861 ttgcgacacc ctccctggtg agcgcgtctt tgtcccgcgt ttgtgaggca ggaagaatat
     7921 acaagttttc ggttcgttta tcttttataa gtgcctgatt aagagtcgcg tcgccttgga
     7981 tgacattgac aaaatcgtag acaacgcgcc gttcgcagcc cattatcagg tccaggttcc
     8041 tgagcccaat atcaaaatct atgaccacag ttttcttgcc tttctgtgcg aggccggtag
     8101 ctatggctgc gctggatgtg gtttttccaa ctccgccctt acctgaagtc acaactatta
     8161 tgcgggccat ggtacccgca gagccagcac ttccggcgct gtccttgtac agctcgtcca
     8221 tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg ttccacgatg gtgtagtcct
     8281 cgttgtggga ggtgatgtcc aacttgatgt tgacgttgta ggcgccgggc agctgcacgg
     8341 gcttcttggc cttgtaggtg gtcttgacct cagcgtcgta gtggccgccg tccttcagct
     8401 tcagcctctg cttgatctcg cccttcaggg cgccgtcctc ggggtacatc cgctcggagg
     8461 aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg aagttggtgc
     8521 cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg
     8581 tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
     8641 acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca
     8701 tgaactgagg ggacaggatg tcccaggcga agggcagggg gccacccttg gtcaccttca
     8761 gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact
     8821 cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc ttgatgatgg
     8881 ccatgttatc ctcctcgccc ttgctcacca tctgctttga gattcgtcgg aacacatgat
     8941 aatagaggtc ccaggcttgg gtgaggtcct tgacattccc tgatttcatg tacttcctgc
     9001 accactcttg ggcctccatt aaatctcgac cataggcctg attaaaggat gtttccttca
     9061 gagtctgggg gccccgttcc atcatagcat gcaagggctc cagcacctca aacatgcctt
     9121 tcacgttcct ttccccaaag tacaaacgag atgcctcttc caggccttca tgccacatct
     9181 ccatggtggc ggatcctcac gacacctgaa atggaagaaa aaaactttga accactgtct
     9241 gaggcttgag aatgaaccaa gatccaaact caaaaagggc aaattccaag gagaattaca
     9301 tcaagtgcca agctggccta acttcagtct ccacccactc agtgtgggga aactccatcg
     9361 cataaaaccc ctccccccaa cctaaagacg acgtactcca aaagctcgag aactaatcga
     9421 ggtgcctgga cggcgcccgg tactccgtgg agtcacatga agcgacggct gaggacggaa
     9481 aggccctttt cctttgtgtg ggtgactcac ccgcccgctc tcccgagcgc cgcgtcctcc
     9541 attttgagct ccctgcagca gggccgggaa gcggccatct ttccgctcac gcaactggtg
     9601 ccgaccgggc cagccttgcc gcccagggcg gggcgataca cggcggcgcg aggccaggca
     9661 ccagagcagg ccggccagct tgagactacc cccgtccgat tctcggtggc cgcgctcgca
     9721 ggccccgcct cgccgaacat gtgcgctggg acgcacgggc cccgtcgccg cccgcggccc
     9781 caaaaaccga aataccagtg tgcagatctt ggcccgcatt tacaagacta tcttgccaga
     9841 aaaaaagcgt cgcagcaggt catcaaaaat tttaaatggc tagagactta tcgaaagcag
     9901 cgagacaggc gcgaaggtgc caccagattc gcacgcggcg gccccagcgc ccaggccagg
     9961 cctcaactca agcacgaggc gaaggggctc cttaagcgca aggcctcgaa ctctcccacc
    10021 cacttccaac ccgaagctcg ggatcaagaa tcacgtactg cagccaggtg gaagtaattc
    10081 aaggcacgca agggccataa cccgtaaaga ggccaggccc gcgggaacca cacacggcac
    10141 ttacctgtgt tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac
    10201 ttatatacgg ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt
    10261 cccagtttac cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca
    10321 acttctcggg gactgtgggc gatgtgcgct ctgcccactg acgggcacct cgagtgaaga
    10381 cgaaagggcc tcatcgataa actggatctc tgctgtccct gtaataaacc cgaaaatttt
    10441 gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta
    10501 ctattctttc ccctgcactg tacccc
//