LOCUS       sequence-243862-        9322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tuning expression of EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9322)
  AUTHORS   Michaels YS, Barnkob MB, Barbosa H, Baeumler TA, Thompson MK, Andre 
            V, Colin-York H, Fritzsche M, Gileadi U, Sheppard HM, Knapp DJHF, 
            Milne TA, Cerundolo V, Fulga TA
  TITLE     Precise tuning of gene expression levels in mammalian cells.
  JOURNAL   Nat Commun. 2019 Feb 18;10(1):818. doi: 10.1038/s41467-019-08777-y.
  PUBMED    30778069
REFERENCE   2  (bases 1 to 9322)
  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: "2019-02-18"; volume: "10(1):818. doi"; pages: " 
            10.1038/s41467-019-08777-y"
COMMENT     Sequence Label: EGFP-miSFIT-9-G
FEATURES             Location/Qualifiers
     source          1..9322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1751..2206)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1888..1909)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2100..2119
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2333..2355
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2347..2364
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2348..2364
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2374..2393
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2374..2392
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      2413..2548
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2588..2709
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             2781..3014
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3101..3689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3207..3218
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3616..3636
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     promoter        complement(4693..5203)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4756..4774)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     promoter        5231..5269
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(5367..5369,5370..5372,5373..6086)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5367..5369,5370..5372,5373..6083)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5412..5433)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5673..5692)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6020..6041
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6221..6355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    6492..6609
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(6876..6920)
                     /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    7105..7338
                     /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    7831..7956
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             8003..8183
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(8198..8401)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        8402..8781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(8882..8902)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8882..8900)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8921..8937)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8921..8937)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8934..8956)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8945..8961
                     /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(8969..8975,8976..8993,8994..8999))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9014..9035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9152..9169)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     1801 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     1861 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     1921 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     1981 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     2041 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2101 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2161 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
     2221 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     2281 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     2341 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     2401 ggtacgtccc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     2461 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     2521 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggata gctagagcca
     2581 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     2641 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     2701 aaacaagttc ctctcactct ctgatattca tttctttgca agttataaat actgaataat
     2761 aagatgacat gaactactac tgctagagat tttccacact gactaaaagg gtctgaggga
     2821 tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa
     2881 gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat
     2941 ctggtctaac cagagagacc cagtacaagc aaaaagcaga tcttgtcttc gttgggagtg
     3001 aattagccct tccagtcccc ccttttcttt taaaaagtgg ctaagatcta cagctgcctt
     3061 gtaagtcatt ggtcttaaag gtaccgagct cgaattccag gcggggaggc ggcccaaagg
     3121 gagatccgac tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc
     3181 aggccgcggg aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc
     3241 ccgcgcagaa tccaggtggc aacacaggcg agcagccatg gaaaggacgt cagcttcccc
     3301 gacaacacca cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg
     3361 caggcggcga tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg
     3421 agctgacagg tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac
     3481 accacgccac gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat
     3541 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
     3601 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
     3661 tcacaaattt tgtaatccag aggttgattg tcgactctag actcgactct agaggatccc
     3721 cctgttccac ctcttgaagg ctatgtaggc cacaaggccc acaaccacag cagccaggat
     3781 ggagcaatag acagggatga ggttgtcggt ggtgcctcgg gtcaccacgg gctgggagct
     3841 gcccatcact gtggtcacca cacctgccac cgtgctggct atgaggtctt gttctggagg
     3901 tgcctcaggc tcctgggtgc tgggggctgt gctgtccgag ccctctgggg gtgtggaccg
     3961 tgtaatccaa cggccaggga tctcctcgca ctcggcgtcg gcccagcgtg tgcactcgcg
     4021 gagctggcgc tcggtgtcct cgcacacggt gcagggcagg cacgggtcca cgtggttggc
     4081 ctcgtcggaa tacgtgccgt cggggcactc ctcgcacacg gtgttctgct tgtcctggca
     4141 ggagaacacg aggcccgagc ccgcctcgca cacgcggcac gcctcgcagc gcccagtcgt
     4201 ctcatcctgg tagtagccgt aggcgcagcg gcacacggcg tcgtcggcct ccacgcacgg
     4261 cgccgacatg ctctggagcc ccacgcactc ggtgcacggc ttgcacggct cggtcgcgct
     4321 caccacgtcg gagaacgtca cgctgtccag gcagggctca cacacggtct ggttggctcc
     4381 acaaggctgg gccacaccct cgcccaggtt gcaggctttg cagcactcac cgctgtgtgt
     4441 gtacaggcct gtggggcatg cctccttggc acctccaagg gacaccccca gaagcagcaa
     4501 cagcagcagg cgcggcccgt ccatggcgcg gccggtggca cctgccccca tcgcccgcct
     4561 cccgcggcag cgctcgactt ccagctcggt ccgctttgcg gactgatggg gctgcgctgc
     4621 gctgcgctcc agcgcccccc ctgcccgccg gagctggccg cggcccgaat tcctgcagcc
     4681 cgggggatcc ccctggggag agaggtcggt gattcggtca acgagggagc cgactgccga
     4741 cgtgcgctcc ggaggcttgc agaatgcgga acaccgcgcg ggcaggaaca gggcccacac
     4801 taccgcccca caccccgcct cccgcaccgc cccttcccgg ccgctgctct cggcgcgccc
     4861 tgctgagcag ccgctattgg ccacagccca tcgcggtcgg cgcgctgcca ttgctccctg
     4921 gcgctgtccg tctgcgaggg tactagtgag acgtgcggct tccgtttgtc acgtccggca
     4981 cgccgcgaac cgcaaggaac cttcccgact taggggcgga gcaggaagcg tcgccggggg
     5041 gcccacaagg gtagcggcga agatccgggt gacgctgcga acggacgtga agaatgtgcg
     5101 agacccaggg tcggcgccgc tgcgtttccc ggaaccacgc ccagagcagc cgcgtccctg
     5161 cgcaaaccca gggctgcctt ggaaaaggcg caaccccaac cccgtggaat tcgatatcaa
     5221 gcttctcgag ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa
     5281 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
     5341 ccgatccagc ctccgcggaa ttcaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     5401 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     5461 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     5521 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     5581 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     5641 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     5701 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     5761 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     5821 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     5881 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     5941 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     6001 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     6061 tcggcatgga cgagctgtac aagtaaagcg gccgcgtcga gagatggggg aggctaactg
     6121 agctagccta cctgcgctgt aagcactttg accggtacta gcttagtcat gcaccgggct
     6181 gcaggaattc gatgacaggc gcagcttccg agggatttga gatccagaca tgataagata
     6241 cattgatgag tttggacaaa ccaaaactag aatgcagtga aaaaaatgcc ttatttgtga
     6301 aatttgtgat gctattgcct tatttgtaac cattataagc tgcaataaac aagttaacaa
     6361 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     6421 caagtaaaac ctctacaaat gtggtatggc tgattatgat cagatctctc gaggctagtc
     6481 tcgtgatcga taaaattttg aatttttgta atttgttttt gtaattcttt agtttgtatg
     6541 tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt
     6601 cttttaaaag ttaaccgata ccgtcgagat ccgttcacta atcgaatgga tctgtctctg
     6661 tctctctctc caccttcttc ttctattcct tcgggcctgt cgggtcccct cggggttggg
     6721 aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac tatagaaagt
     6781 acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa taattttata
     6841 taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt atctaattcc
     6901 aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa ggagtgtatt
     6961 aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg attccaaatc
     7021 tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt gcaaatgagt
     7081 tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct ttccacagcc
     7141 aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag atgctgttgc
     7201 gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa taattgtctg
     7261 gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc tcccaagaac
     7321 ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac tcttctcttt
     7381 gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt atataattca
     7441 cttctccaat tgtccctcat atctcctcct ccaggtctga agatcagcgg ccgcttgctg
     7501 tgcggtggtc ttacttttgt tttgctcttc ctctatcttg tctaaagctt ccttggtgtc
     7561 ttttatctct atcctttgat gcacacaata gagggttgct actgtattat ataatgatct
     7621 aagttcttct gatcctgtct gaagggatgg ttgtagctgt cccagtattt gtctacagcc
     7681 ttctgatgtt tctaacaggc caggattaac tgcgaatcgt tctagctccc tgcttgccca
     7741 tactatatgt tttaatttat attttttctt tccccctggc cttaaccgaa ttttttccca
     7801 tcgcgatcta attctccccc gcttaatact gacgctctcg cacccatctc tctccttcta
     7861 gcctccgcta gtcaaaattt ttggcgtact caccagtcgc cgcccctcgc ctcttgccgt
     7921 gcgcgcttca gcaagccgag tcctgcgtcg agagagctcc tctggtttcc ctttcgcttt
     7981 caagtccctg ttcgggcgcc actgctagag attttccaca ctgactaaaa gggtctgagg
     8041 gatctctagt taccagagtc acacaacaga cgggcacaca ctacttgaag cactcaaggc
     8101 aagctttatt gaggcttaag cagtgggttc cctagttagc cagagagctc ccaggctcag
     8161 atctggtcta accagagaga ccccggttca ctaaacgagc tctgcttata tagacctccc
     8221 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     8281 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     8341 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     8401 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     8461 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     8521 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     8581 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     8641 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     8701 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     8761 aactagtcaa taatcaatgt caacatggcg gtaatgttgg acatgagcca atataaatgt
     8821 acatattatg atatggatac aacgtatgca atggccaagc ttgcagctcc agcttttgtt
     8881 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     8941 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     9001 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     9061 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     9121 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     9181 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     9241 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     9301 aaaaggccgc gttgctggcg tt
//
