LOCUS       sequence-416580-       15323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To monitor the status of EGF, the plasmid encodes a recombinant EGF 
            fused extracellularly to the mScarlet fluorescent protein and 
            intracellularly to the mNeonGreen fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15323)
  AUTHORS   Deguchi E, Lin S, Hirayama D, Matsuda K, Tanave A, Sumiyama K, 
            Tsukiji S, Otani T, Furuse M, Sorkin A, Matsuda M, Terai K
  TITLE     Low-affinity ligands of the epidermal growth factor receptor are 
            long-range signal transmitters in collective cell migration of 
            epithelial cells.
  JOURNAL   Cell Rep. 2024 Nov 14;43(11):114986. doi: 
            10.1016/j.celrep.2024.114986.
  PUBMED    39546398
REFERENCE   2  (bases 1 to 15323)
  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: "Cell Rep.";
            date: "2024-11-14"; volume: "43(11):114986. doi"; pages: " 
            10.1016/j.celrep.2024.114986"
COMMENT     Sequence Label: pCSIIpuro-EGF-ScNeo
FEATURES             Location/Qualifiers
     source          1..15323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        616..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             832..1012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1059..1184
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1676..1909
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2094..2138
                     /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    2412..2529
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2585..3747
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2816..3738
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3695..3715
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             6562..6588
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6589..7284
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(6722..6740)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6990..7009
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    8832..9381
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8976..8993)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     9203..9222
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             9384..9983
                     /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(9384..9403)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9880..9900
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10018..10606
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10071..10091)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10489..10500)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11125..11305
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11331..11348)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11337..11561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11607..12035
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11694..11713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11904..11925
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12044..12064)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12049..12378
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12229..12364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12291..12310
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12426..12473
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12492..12866
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    12996..13117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13033..13052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13087..13106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13166..13182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13166..13182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13179..13201)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13190..13206
                     /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(13214..13220,13221..13238,13239..13244))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13259..13280
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13397..13414)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13568..14156)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13648..13667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14327..15118,15119..15187))
                     /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     14950..14969
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15188..15292)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc
      841 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      901 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      961 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     1021 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     1081 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     1141 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     1201 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     1261 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     1321 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     1381 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     1441 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     1501 gtaagaccac cgcacagcaa gggccgctga tcttcagacc tggaggagga gatatgaggg
     1561 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1621 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1681 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1741 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1801 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1861 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1921 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1981 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     2041 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     2101 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     2161 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     2221 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     2281 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     2341 gtggagagag agacagagac agatccattc gattagtgaa cggatctaca aatggcagta
     2401 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     2461 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     2521 caaaattttc gggtttatta cagggacagc agaaattcac tttgaattaa ttcaagcttc
     2581 gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     2641 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
     2701 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     2761 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     2821 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     2881 aattacttcc acctggctcc agtacgtgat tcttgatccc gagctggagc caggggcggg
     2941 ccttgcgctt taggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
     3001 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     3061 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     3121 gtaaatgcgg gccaggatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     3181 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     3241 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     3301 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     3361 gaaagatggc cgcttcccgg ccctgctcca gggggctcaa aatggaggac gcggcgctcg
     3421 ggagagcggg cgggtgagtc acccacacaa aggaaagggg cctttccgtc ctcagccgtc
     3481 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctggagc
     3541 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     3601 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     3661 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     3721 ttttttcttc catttcaggt gtcgtgaaca cgctaccggt ccgaattcac catgctgctc
     3781 actcttatca ttctgttgcc agtagtttca aaatttagtt ttgttagtct ctcagcaccg
     3841 cagcactgga gctgtcctga aggtactctc gcaggaaatg ggaattctac ttgtgtgggt
     3901 cctgcaccct tcttaatttt ctcccatgga aatagtatct ttaggattga cacagaagga
     3961 accaattatg agcaattggt ggtggatgct ggtgtctcag tgatcatgga ttttcattat
     4021 aatgagaaaa gaatctattg ggtggattta gaaagacaac ttttgcaaag agtttttctg
     4081 aatgggtcaa ggcaagagag agtatgtaat atagagaaaa atgtttctgg aatggcaata
     4141 aattggataa atgaagaagt tatttggtca aatcaacagg aaggaatcat tacagtaaca
     4201 gatatgaaag gaaataattc ccacattctt ttaagtgctt taaaatatcc tgcaaatgta
     4261 gcagttgatc cagtagaaag gtttatattt tggtcttcag aggtggctgg aagcctttat
     4321 agagcagatc tcgatggtgt gggagtgaag gctctgttgg agacatcaga gaaaataaca
     4381 gctgtgtcat tggatgtgct tgataagcgg ctgttttgga ttcagtacaa cagagaagga
     4441 agcaattctc ttatttgctc ctgtgattat gatggaggtt ctgtccacat tagtaaacat
     4501 ccaacacagc ataatttgtt tgcaatgtcc ctttttggtg accgtatctt ctattcaaca
     4561 tggaaaatga agacaatttg gatagccaac aaacacactg gaaaggacat ggttagaatt
     4621 aacctccatt catcatttgt accacttggt gaactgaaag tagtgcatcc acttacacaa
     4681 cccaaggcag aagatgacac ttgggagcct gatgttaatg aatgtgcttt ttggaatcat
     4741 ggctgtactc ttgggtgtaa aaacacccct ggatcctatt actgcacgtg ccctgtagga
     4801 tttgttctgc ttcctgatgg gaaacgatgt catcaacttg tttcctgtcc acgcaatgtg
     4861 tctgaatgca gccatgactg tgttctgaca tcagaaggtc ccttatgttt ctgtcctgaa
     4921 ggctcagtgc ttgagagaga tgggaaaaca tgtagcggtt gttcctcacc cgataatggt
     4981 ggatgtagcc agctctgcgt tcctcttagc ccagtatcct gggaatgtga ttgctttcct
     5041 gggtatgacc tacaactgga tgaaaaaagc tgtgcagctt caggaccaca accatttttg
     5101 ctgtttgcca attctcaaga tattcgacac atgcattttg atggaacaga ctatggaact
     5161 ctgctcagcc agcagatggg aatggtttat gccctagatc atgaccctgt ggaaaataag
     5221 atatactttg cccatacagc cctgaagtgg atagagagag ctaatatgga tggttcccag
     5281 cgagaaaggc ttattgagga aggagtagat gtgccagaag gtcttgctgt ggactggatt
     5341 ggccgtagat tctattggac agacagaggg aaatctctga ttggaaggag tgatttaaat
     5401 gggaaacgtt ccaaaataat cactaaggag aacatctctc aaccacgagg aattgctgtt
     5461 catccaatgg ccaagagatt attctggact gatacaggga ttaatccacg aattgaaagt
     5521 tcttccctcc aaggccttgg ccgtctggtt atagccagct ctgatctaat ctggcccagt
     5581 ggaataacga ttgacttctt aactgacaag ttgtactggt gcgatgccaa gcagtctgtg
     5641 attgaaatgg ccaatctgga tggttcaaaa cgccgaagac ttacccagaa tgatgtaggt
     5701 cacccatttg ctgtagcagt gtttgaggat tatgtgtggt tctcagattg ggctatgcca
     5761 tcagtaataa gagtaaacaa gaggactggc aaagatagag tacgtctcca aggcagcatg
     5821 ctgaagccct catcactggt tgtggttcat ccattggcaa aaccaggagc agatccctgc
     5881 ttatatcaaa acggaggctg tgaacatatt tgcaaaaaga ggcttggaac tgcttggtgt
     5941 tcgtgtcgtg aaggttttat gaaagcctca gatgggaaaa cgtgtctggc tctggatggt
     6001 catcagctgt tggcaggtgg tgaagttgat ctaaagaacc aagtaacacc attggacatc
     6061 ttgtccaaga ctagagtgtc agaagataac attacagaat ctcaacacat gctagtggct
     6121 gaaatcatgg tgtcagatca agatgactgt gctcctgtgg gatgcagcat gtatgctcgg
     6181 tgtatttcag agggagagga tgccacatgt cagtgtttga aaggatttgc tggggatgga
     6241 aaactatgtt ctgatataga tgaatgtgag atgggtgtcc cagtgtgccc ccctgcctcc
     6301 tccaagtgca tcaacaccga aggtggttat gtctgccggt gctcagaagg ctaccaagga
     6361 gatgggattc actgtcttga tattgatgag tgccaactgg gggtgcacag ctgtggagag
     6421 aatgccagct gcacaaatac agagggaggc tatacctgca tgtgtgctgg acgcctgtct
     6481 gaaccaggac tgatttgccc tgactctact ccaccccctc acctcaggga agatgaccac
     6541 cactattccg taagaaatag ttacccatac gatgttccgg attacgctat ggtgagcaag
     6601 ggcgaggcag tgatcaagga gttcatgcgg ttcaaggtgc acatggaggg ctccatgaac
     6661 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     6721 gccaagctga aggtgaccaa gggtggcccc ctgcccttct cctgggacat cctgtcccct
     6781 cagttcatgt acggctccag ggccttcacc aagcaccccg ccgacatccc cgactactat
     6841 aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     6901 gccgtgaccg tgacccagga cacctccctg gaggacggca ccctgatcta caaggtgaag
     6961 ctccgcggca ccaacttccc tcctgacggc cccgtaatgc agaagaagac aatgggctgg
     7021 gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc tgaagggcga cattaagatg
     7081 gccctgcgcc tgaaggacgg aggccgctac ctggcggact tcaagaccac ctacaaggcc
     7141 aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc gcaagttgga catcacctcc
     7201 cacaacgagg actacaccgt ggtggaacag tacgaacgct ccgagggccg ccactccacc
     7261 ggcggcatgg acgagctgta caagggtggt ggtagtggtg gtggttcaaa tagtgactct
     7321 gaatgtcccc tgtcccacga tgggtactgc ctccatgatg gtgtgtgcat gtatattgaa
     7381 gcattggaca agtatgcatg caactgtgtt gttggctaca tcggggagcg atgtcagtac
     7441 cgagacctga agtggtggga actgcgccac gctggccacg ggcagcagca gaaggtcatc
     7501 gtggtggctg tctgcgtggt ggtgcttgtc atgctgctcc tcctgagcct gtggggggcc
     7561 cactactaca ggactcagaa gctgctatcg aaaaacccaa agaatcctta tgaggagtcg
     7621 agcagagatg tgaggagtcg caggcctgct gacactgagg atgggatgtc ctcttgccct
     7681 caaccttggt ttgtggttat aaaagaacac caagacctca agaatggggg tcaaccagtg
     7741 gctggtgagg atggccaggc agcagatggg tcaatgcaac caacttcatg gaggcaggag
     7801 ccccagttat gtggaatggg cacagagcaa ggctgctgga ttccagtatc cagtgataag
     7861 ggctcctgtc cccaggtaat ggagcgaagc tttcatatgc cctcctatgg gacacagacc
     7921 cttgaagggg gtgtcgagaa gccccattct ctcctatcag ctaacccatt atggcaacaa
     7981 agggccctgg acccaccaca ccaaatggag ctgactcagg gcggattcga aatggtaagc
     8041 aagggggagg aagataatat ggcttccctt ccagcaacgc acgaattgca tatctttgga
     8101 agtattaatg gagttgattt cgacatggtg gggcagggta cgggaaaccc aaacgatggc
     8161 tatgaagaac ttaacctcaa gtctaccaaa ggcgatctcc aattctcacc ctggatattg
     8221 gtcccacaca tcggctacgg gttccatcaa tacctgcctt atcctgacgg gatgagtcct
     8281 tttcaagctg ctatggttga cggaagtggc taccaagttc accgcacaat gcaatttgag
     8341 gatggtgcat ctcttacggt taattaccga tatacatatg agggttcaca catcaagggg
     8401 gaagctcagg tcaagggcac tggatttccc gcagatggcc cagttatgac caattctctg
     8461 accgcagccg attggtgtcg aagcaagaag acctacccta acgacaagac tatcatctca
     8521 accttcaaat ggtcctacac cacaggtaac ggaaagcggt ataggtctac agctcggaca
     8581 acatatacct ttgcgaagcc tatggccgct aactatttga aaaaccaacc gatgtatgtt
     8641 tttcggaaga ccgaattgaa acattcaaaa acggagttga atttcaaaga gtggcagaag
     8701 gcattcaccg acgtaatggg tatggatgag ttgtataaat aagcggcggc cgcggctcta
     8761 gaggatccgt taactaactt aagctagcaa cggtttccct ctagcgggat caattccgcc
     8821 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     8881 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     8941 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     9001 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     9061 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     9121 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     9181 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     9241 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     9301 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     9361 cctttgaaaa acacgataat accatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     9421 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     9481 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     9541 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     9601 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     9661 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     9721 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     9781 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     9841 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     9901 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     9961 tgacccgcaa gcccggtgcc tgactagagt taacatcgag ggatcaagct tatcgataat
    10021 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
    10081 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
    10141 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
    10201 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
    10261 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
    10321 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
    10381 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
    10441 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
    10501 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
    10561 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga
    10621 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
    10681 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
    10741 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
    10801 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
    10861 cacacacaag gctacttccc tgattggcag aactacacac cagggccagg gatcagatat
    10921 ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa
    10981 gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac
    11041 ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc
    11101 cgagagctgc atccggactg tactgggtct ctctggttag accagatctg agcctgggag
    11161 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
    11221 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
    11281 tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt
    11341 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
    11401 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
    11461 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
    11521 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
    11581 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
    11641 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
    11701 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
    11761 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
    11821 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
    11881 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
    11941 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
    12001 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
    12061 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
    12121 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
    12181 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
    12241 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
    12301 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
    12361 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
    12421 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
    12481 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
    12541 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
    12601 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
    12661 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
    12721 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
    12781 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
    12841 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
    12901 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
    12961 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
    13021 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
    13081 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
    13141 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
    13201 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
    13261 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
    13321 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
    13381 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
    13441 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
    13501 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
    13561 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
    13621 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
    13681 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
    13741 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
    13801 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
    13861 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
    13921 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
    13981 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
    14041 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
    14101 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
    14161 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
    14221 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
    14281 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
    14341 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
    14401 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
    14461 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
    14521 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
    14581 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
    14641 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
    14701 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
    14761 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
    14821 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
    14881 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
    14941 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
    15001 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
    15061 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
    15121 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    15181 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    15241 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    15301 cccgaaaagt gccacctgac gtc
//
