LOCUS       sequence-461394-       10418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of N581K RLIM-GFP-IRES-mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10418)
  AUTHORS   Bandi V, Rennie M, Koch I, Gill P, Pacheco OD, Berg AD, Cui H, Ward 
            DI, Bustos F
  TITLE     RLIM-specific activity reporters define variant pathogenicity in 
            Tonne-Kalscheuer syndrome.
  JOURNAL   HGG Adv. 2024 Oct 31;6(1):100378. doi: 10.1016/j.xhgg.2024.100378.
  PUBMED    39482882
REFERENCE   2  (bases 1 to 10418)
  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: "HGG Adv."; 
            date: "2024-10-31"; volume: "6(1):100378. doi"; pages: " 
            10.1016/j.xhgg.2024.100378"
COMMENT     Sequence Label: pCAGGS puro human-GFP-N581K RLIM-IRES-mCherry
FEATURES             Location/Qualifiers
     source          1..10418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     448..467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     701..718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    835..856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(871..876,877..894,895..901)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    909..925
                     /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     914..936
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     933..949
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     933..949
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     979..998
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        979..997
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        1007..1386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1388..1662
                     /label=chicken beta-actin promoter
     intron          1663..2679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2603..2625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2687..2706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2742..3458
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     misc_feature    5361..5933
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5528..5545)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5755..5774
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5942..6652
                     /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(6087..6105)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     polyA_signal    7013..7147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7050..7069)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7104..7123
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        7152..7651
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7187..7210)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7584..7603
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7667..8266
                     /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(7667..7686)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8163..8183
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(8602..8619)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8602..8618)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8611..8633)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(8759..9214)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8896..8917)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9108..9127
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        9241..9345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9346..9414,9415..10206)
                     /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(9564..9583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10377..10418,1..547)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
       61 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
      601 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
      661 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
      721 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
      781 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
      841 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      901 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa
      961 tttggccaag tcggcctcta atacgactca ctatagggag ctcgtcgaca ttgattattg
     1021 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     1081 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     1141 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     1201 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     1261 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     1321 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     1381 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
     1441 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
     1501 ggggggggcg cgccaggsgg ggsggggsgg ggsgaggggs ggggsggggs gaggcggaga
     1561 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
     1621 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
     1681 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
     1741 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
     1801 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
     1861 gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg
     1921 agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
     1981 ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc
     2041 ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca
     2101 gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct
     2161 gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg
     2221 ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc kcgggccggg
     2281 grgggctcgg gggrggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg
     2341 agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc
     2401 caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     2461 gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg
     2521 cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt
     2581 cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc
     2641 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     2701 attgtgctgt ctcatcattt tggcaaagaa ttcaactcga gatggtgagc aagggcgccg
     2761 agctgttcac cggcatcgtg cccatcctga tcgagctgaa tggcgatgtg aatggccaca
     2821 agttcagcgt gagcggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
     2881 tcatctgcac caccggcaag ctgcctgtgc cctggcccac cctggtgacc accctgagct
     2941 acggcgtgca gtgcttctca cgctaccccg atcacatgaa gcagcacgac ttcttcaaga
     3001 gcgccatgcc tgagggctac atccaggagc gcaccatctt cttcgaggat gacggcaact
     3061 acaagtcgcg cgccgaggtg aagttcgagg gcgataccct ggtgaatcgc atcgagctga
     3121 ccggcaccga tttcaaggag gatggcaaca tcctgggcaa taagatggag tacaactaca
     3181 acgcccacaa tgtgtacatc atgaccgaca aggccaagaa tggcatcaag gtgaacttca
     3241 agatccgcca caacatcgag gatggcagcg tgcagctggc cgaccactac cagcagaata
     3301 cccccatcgg cgatggccct gtgctgctgc ccgataacca ctacctgtcc acccagagcg
     3361 ccctgtccaa ggaccccaac gagaagcgcg atcacatgat ctacttcggc ttcgtgaccg
     3421 ccgccgccat cacccacggc atggatgagc tgtacaaggg aggcggaggc ggatccatgg
     3481 aaaactcaga ttccaatgac aaaggaagtg gtgatcagtc tgcagcacag cgcagaagtc
     3541 agatggaccg attggatcga gaagaagctt tctatcaatt tgtaaataac ctgagtgaag
     3601 aagattatag gcttatgaga gataacaatt tgctaggcac cccaggtgaa agtactgagg
     3661 aagagttgct gagacgacta cagcaaatta aagaaggccc accaccgcaa aactcagatg
     3721 aaaatagagg aggagactct tcagatgatg tgtctaatgg tgactctata atagactggc
     3781 ttaactctgt cagacaaact ggaaatacaa caagaagtgg gcaaagagga aaccaatctt
     3841 ggagagcagt gagtcggact aatccaaaca gtggtgattt cagattcagt ttagagataa
     3901 atgttaaccg taataatggg agccaaaatt cagagaatga aaatgagcca tctgcaagac
     3961 gttctagtgg agaaaatgtg gaaaacaaca gccaaaggca agtggaaaac ccacgatctg
     4021 aatcaacatc tgcaaggcca tctagatcag aacgaaattc aactgaagcg ttaacagagg
     4081 tcccacctac cagaggtcag aggagggcaa gaagcaggag cccagaccat cggagaacca
     4141 gagcaagagc tgaaagaagt aggtcacctc tgcatccaat gagtgaaatt ccacgaagat
     4201 ctcatcatag tatctcatct cagacttttg aacatccttt ggtaaatgag acggagggaa
     4261 gttctagaac ccggcaccat gtgacattga ggcagcaaat atctgggcct gagttgctaa
     4321 gtagaggtct ttttgcagct tctggaacaa gaaatgcttc tcaaggagca ggttcttcag
     4381 acacagctgc cagtggtgaa tctacaggat caggacagag acctccaacc atagtccttg
     4441 atcttcaagt aagaagagtt cgtcctggag aatatcggca gagagatagc atagccagca
     4501 gaactcggtc taggtctcag acaccaaaca acactgtcac ctatgaaagt gaacgaggag
     4561 gttttaggcg tacattttca cgttctgagc gggcaggtgt gagaacctat gtcagtacca
     4621 tcagaattcc cattcgtaga atcttaaata ctggtttaag tgagactaca tctgttgcaa
     4681 ttcagaccat gttaaggcag ataatgacag gttttggtga gttaagctat tttatgtaca
     4741 gtgatagcga ctcagagcct actggctcag tctcaaatcg aaatatggaa agggcagagt
     4801 cacggagtgg aagaggaggt tctggtggtg gtagtagttc tggttccagt tcgagttcca
     4861 gttccagttc gagttccagt tccagttcaa gttccagttc cagtcctagt tccagttccg
     4921 gtggtgaaag ttcagaaact agctcagatt tatttgaagg cagtaatgaa ggaagctcat
     4981 catcaggctc atcaggtgcc aggcgagagg gtcgacatag ggccccagtc acatttgatg
     5041 aaagtggctc tttgcccttc cttagcctgg ctcagttttt cctcttaaat gaggatgatg
     5101 atgaccaacc tagaggactc accaaagaac agattgacaa cttggcaatg agaagttttg
     5161 gtgaaaatga tgcattaaaa acctgtagtg tttgcattac agaatataca gaaggcaaga
     5221 aacttcgtaa actaccttgt tcccatgagt accatgtcca ctgcatcgat cgctggttat
     5281 ctgagaattc tacctgtcct atttgtcgca gagcagtctt agcttctggt aacagagaaa
     5341 gtgttgtgta aggatcctag cccctctccc tccccccccc ctaacgttac tggccgaagc
     5401 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     5461 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     5521 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     5581 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     5641 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     5701 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     5761 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     5821 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     5881 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata atacctccgg
     5941 aatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     6001 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     6061 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     6121 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     6181 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     6241 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     6301 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     6361 aatgcagaag aaaaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     6421 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     6481 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     6541 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     6601 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt gactcgagcc
     6661 ttaattaagg gctagcaaat gcatgcggcc gccaattcca gctgagcgcc ggtcgctacc
     6721 attaccagtt ggtctggtgt caaaaataat aataaccggg caggccatgt ctgcccgtat
     6781 ttcgcgtaag gaaatccatt atgtactatt taaaaaacac aaacttttgg atgttcggtt
     6841 tattcttttt cttttacttt tttatcatgg gagcctactt cccgtttttc ccgatttggc
     6901 tacatgacat caaccatatc agcaaaagtg atacgggtat tatttttgcc gctatttctc
     6961 tgttctcgct attattccaa ccgctgtttg gtctgctttc tgacaaactc ggaacttgtt
     7021 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     7081 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     7141 ctggatctac cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc
     7201 agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc
     7261 caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc
     7321 tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat
     7381 ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg
     7441 ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag
     7501 gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc
     7561 ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct
     7621 gttctcctct tcctcatctc cgggcctttc gacaagctag cttaccatga ccgagtacaa
     7681 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     7741 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     7801 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     7861 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     7921 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     7981 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     8041 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     8101 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     8161 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     8221 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc cgccccacga
     8281 cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca tcgatttgcc agccatctgt
     8341 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     8401 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     8461 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     8521 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga cgcggccggc
     8581 cgctgaccaa gtcagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     8641 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     8701 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggaa
     8761 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     8821 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     8881 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     8941 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     9001 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     9061 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     9121 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     9181 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     9241 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     9301 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     9361 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     9421 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     9481 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     9541 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     9601 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     9661 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     9721 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     9781 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     9841 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     9901 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     9961 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
    10021 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
    10081 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
    10141 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
    10201 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
    10261 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
    10321 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
    10381 gatccttttt ttctgcgcgt aatctgctgc ttgcaaac
//
