LOCUS       sequence-424333-        8996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FLAG epitope-tagged OgNLuc ""Orange"" BRET reporter in 
            mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8996)
  AUTHORS   Labra B, Parag-Sharma K, Powers JJ, Srivastava S, Walker JR, 
            Kirkland TA, Brennan CK, Prescher JA, Amelio AL
  TITLE     Optimized in vivo multispectral bioluminescent imaging of tumor 
            biology using engineered BRET reporters.
  JOURNAL   iScience. 2024 Aug 8;27(9):110655. doi: 10.1016/j.isci.2024.110655. 
            eCollection 2024 Sep 20.
  PUBMED    39252965
REFERENCE   2  (bases 1 to 8996)
  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: "iScience.";
            date: "2024-08-8"; volume: "27(9):110655. doi"; pages: " 
            10.1016/j.isci.2024.110655. eCollection 2024 Sep 20"
COMMENT     Sequence Label: pLenti_CMV:Flag-OgNLuc
FEATURES             Location/Qualifiers
     source          1..8996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     190..207
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    324..345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(360..365,366..383,384..390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    398..414
                     /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     403..425
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     422..438
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     422..438
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     457..477
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        459..477
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        505..731
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             732..912
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    959..1084
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1577..1810
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1995..2039
                     /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    2306..2422
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2445..2748
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2749..2952
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2903..2923
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2949..2973
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3032..3055
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3056..3760
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="VSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGF
                     QTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALKG
                     EDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     3466..3485
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3776..4291
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(4334..4354)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4339..4668
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4519..4654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4678..5277
                     /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(4678..4697)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5174..5194
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5344..5932
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5397..5417)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5815..5826)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6019..6252
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6324..6445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6361..6380)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6415..6434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6485..6620
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6640..6659)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6641..6659)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6669..6686)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6669..6685)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6678..6700)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6827..7282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6914..6933)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7124..7145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7308..7412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7413..7481,7482..8273)
                     /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(7631..7650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8444..8996,1..36)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8933..8952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
       61 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      121 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
      181 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
      241 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
      301 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
      361 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
      421 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
      481 caaaagctgg agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca
      541 tggtaacgat gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat
      601 tggtggaagt aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat
      661 ggattggacg aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc
      721 gatacataaa cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
      781 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
      841 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
      901 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
      961 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     1021 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     1081 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     1141 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     1201 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     1261 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     1321 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     1381 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
     1441 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1501 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1561 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1621 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1681 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1741 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1801 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1861 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     1921 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     1981 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     2041 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     2101 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     2161 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     2221 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     2281 acggatctcg acggtatcgg ttaactttta aaagaaaagg ggggattggg gggtacagtg
     2341 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     2401 aaattacaaa attcaaaatt ttatcgataa gcttgggagt tccgcgttac ataacttacg
     2461 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     2521 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     2581 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     2641 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     2701 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     2761 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     2821 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     2881 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     2941 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     3001 gacctccata gaagacaccg actctagaat ggactacaag gacgacgatg acaaggtgag
     3061 caagggcgag gagaataaca tggccatcat caaggagttc atgcgcttca aggtgcgcat
     3121 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     3181 cgagggcttt cagaccgtta agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     3241 ggacatcttg tcccctcagt tcacctacgg ctccaaggcc tacgtgaagc accccgccga
     3301 catccccgac tacctcaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     3361 cttcgaggac ggcggcgtgg tgaccgtgac tcaggactcc tccctgcagg acggcgagtt
     3421 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa
     3481 gaagaccatg ggcatggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     3541 gggcgaggac aagctcaggc tgaagctgaa ggacggcggc cactacacct ccgaggtcaa
     3601 gaccacctac aaggccaaga agcccgtgca gttgcccggc gcctacatcg tcgacatcaa
     3661 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     3721 gggccgccac tccaccggcg gcatggacga gctgtacaag gatatctccg gaggtatggt
     3781 cttcacactc gaagatttcg ttggggactg gcgacagaca gccggctaca acctggacca
     3841 agtccttgaa cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc
     3901 gatccaaagg attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat
     3961 cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt
     4021 gtaccctgtg gatgatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga
     4081 cggggttacg ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt
     4141 cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga
     4201 gcgcctgatc aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg
     4261 ctggcggctg tgcgaacgca ttctggcgta aggatccgga ctagttaacg cgtcgaccag
     4321 tggtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4381 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4441 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4501 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4561 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4621 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaccatg
     4681 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     4741 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     4801 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     4861 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     4921 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     4981 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     5041 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     5101 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     5161 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5221 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc
     5281 ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgccagtc
     5341 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5401 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5461 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5521 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5581 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5641 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5701 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     5761 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5821 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5881 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
     5941 cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     6001 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     6061 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     6121 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6181 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6241 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6301 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     6361 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6421 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     6481 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6541 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6601 aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac
     6661 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     6721 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     6781 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     6841 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6901 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6961 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     7021 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     7081 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     7141 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     7201 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7261 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     7321 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     7381 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     7441 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     7501 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     7561 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     7621 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     7681 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7741 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7801 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7861 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7921 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7981 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     8041 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     8101 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     8161 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     8221 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     8281 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     8341 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     8401 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     8461 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8521 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8581 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8641 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8701 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8761 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8821 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8881 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8941 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgattt
//
