LOCUS       sequence-102370-       14367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3X FLAG fused to wt BirA - mCherry - sec61ß.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14367)
  AUTHORS   Jan CH, Williams CC, Weissman JS
  TITLE     Principles of ER cotranslational translocation revealed by 
            proximity-specific ribosome profiling.
  JOURNAL   Science. 2014 Nov 7;346(6210):1257521. doi: 10.1126/science.1257521.
            Epub 2014 Nov 6.
  PUBMED    25378630
REFERENCE   2  (bases 1 to 14367)
  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: "Science."; 
            date: "2014-11-7"; volume: "346(6210):1257521. doi"; pages: " 
            10.1126/science.1257521. Epub 2014 Nov 6"
COMMENT     Sequence Label: pJW1512
FEATURES             Location/Qualifiers
     source          1..14367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    497..730
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             915..959
                     /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    1257..1374
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1439..1813)
                     /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"
     regulatory      1810..1819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1823..1841)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1833..1880)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    join(1950..1968,1969..1991,1992..2010,2011..2033,2034..2052,
                     2053..2075,2076..2094,2095..2117,2118..2136,2137..2159,
                     2160..2178,2179..2201,2202..2220)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2253..2291
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2288..2312
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2382..2391
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2391..2456
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2460..3422
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2460..3422
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             3432..4136
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3574..3592)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3842..3861
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4149..4436
                     /codon_start=1
                     /gene="SEC61B"
                     /product="human Sec61-beta subunit of the translocon in the
                     endoplasmic reticulum"
                     /label=Sec61-beta
                     /translation="PGPTPSGTNVGSSGRSPSKAVAARAAGSTVRQRKNASCGTRSAGR
                     TTSAGTGGMWRFYTEDSPGLKVGPVPVLVMSLLFIASVFMLHIWGKYTRS"
     promoter        4476..5687
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5524..5543
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    5694..5727
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             join(5734..6351,6352..6357,6358..6393,6394..6396,6397..6432,
                     6433..6435,6436..6471,6472..6480)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     protein_bind    complement(6481..6514)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6530..7102
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6697..6714)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6924..6943
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(7120..7122,7123..7125,7126..7839)
                     /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(7120..7122,7123..7125,7126..7836)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7165..7186)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7426..7445)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7773..7794
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7861..8449
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7914..7934)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8332..8343)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8656..8889
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8990..9007)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8996..9220
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9266..9694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9353..9372)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9563..9584
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9703..9723)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9708..10037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9888..10023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9950..9969
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    10860..10981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11030..11046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11030..11046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11043..11065)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11054..11070
                     /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(11078..11084,11085..11102,11103..11108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11123..11144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11261..11278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11432..12020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11512..11531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12191..12982,12983..13051))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRMDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     12814..12833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13052..13156)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13204..13338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             13506..14139
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ccgagtaata ccgggggaga ttagatcgcg atgggaaaaa ttcggttaag gccaggggga
       61 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
      121 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
      181 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
      241 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
      301 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
      361 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
      421 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
      481 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
      541 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
      601 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
      661 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
      721 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
      781 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
      841 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
      901 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
      961 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     1021 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     1081 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     1141 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     1201 acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt
     1261 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     1321 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     1381 tattacaggg acagcagaga tccagtttgg ttagtaccgg gcccgctcta gtccggaatc
     1441 agtcctgctc ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc agggcgaact
     1501 cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg tcccggaagt
     1561 tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc acccacaccc
     1621 aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac agggtcacgt
     1681 cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac ccgagccggt
     1741 cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga acggcactgg
     1801 tcaacttggc catggtggcc ctcctatagt gagtcgtatt atactatgcc gatatactat
     1861 gccgatgatt aattgtcaac acgtgctgca ggtccgaggt tctaggctag cttaaaggaa
     1921 ccaattcagt cgactggatc ctttaccact ccctatcagt gatagagaaa agtgaaagtc
     1981 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
     2041 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     2101 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     2161 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     2221 aaagtgaaag tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg gaggcctata
     2281 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     2341 acctccatag aagacaccgg gaccgatcca gcctaccggt cgccaccatg gactacaaag
     2401 accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagctta
     2461 tgaaggataa caccgtgcca ctgaaattga ttgccctgtt agcgaacggt gaatttcact
     2521 ctggcgagca gttgggtgaa acgctgggaa tgagccgggc ggctattaat aaacacattc
     2581 agacactgcg tgactggggc gttgatgtct ttaccgttcc gggtaaagga tacagcctgc
     2641 ctgagcctat ccagttactt aatgctaaac agatattggg tcagctggat ggcggtagtg
     2701 tagccgtgct gccagtgatt gactccacga atcagtacct tcttgatcgt atcggagagc
     2761 ttaaatcggg cgatgcttgc attgcagaat accagcaggc tggccgtggt cgccggggtc
     2821 ggaaatggtt ttcgcctttt ggcgcaaact tatatttgtc gatgttctgg cgtctggaac
     2881 aaggcccggc ggcggcgatt ggtttaagtc tggttatcgg tatcgtgatg gcggaagtat
     2941 tacgcaagct gggtgcagat aaagttcgtg ttaaatggcc taatgacctc tatctgcagg
     3001 atcgcaagct ggcaggcatt ctggtggagc tgactggcaa aactggcgat gcggcgcaaa
     3061 tagtcattgg agccgggatc aacatggcaa tgcgccgtgt tgaagagagt gtcgttaatc
     3121 aggggtggat cacgctgcag gaagcgggga tcaatctcga tcgtaatacg ttggcggcca
     3181 tgctaatacg tgaattacgt gctgcgttgg aactcttcga acaagaagga ttggcacctt
     3241 atctgtcgcg ctgggaaaag ctggataatt ttattaatcg cccagtgaaa cttatcattg
     3301 gtgataaaga aatatttggc atttcacgcg gaatagacaa acagggggct ttattacttg
     3361 agcaggatgg aataataaaa ccctggatgg gcggtgaaat atccctgcgt agtgcagaaa
     3421 aaggaggttc tgtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
     3481 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
     3541 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
     3601 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
     3661 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
     3721 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
     3781 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
     3841 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
     3901 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
     3961 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
     4021 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
     4081 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagaaca
     4141 gccgcgtgcc tggtccgacc cccagtggca ctaacgtggg atcctcaggg cgctctccca
     4201 gcaaagcagt ggccgcccgg gcggcgggat ccactgtccg gcagaggaaa aatgccagct
     4261 gtgggacaag gagtgcaggc cgcacaacct cggcaggcac cggggggatg tggcgattct
     4321 acacagaaga ttcacctggg ctcaaagttg gccctgttcc agtattggtt atgagtcttc
     4381 tgttcatcgc ttctgtattt atgttgcaca tttggggcaa gtacactcgt tcgtaggaat
     4441 tcatcgatag atctgatatc ggacgcgtaa gatctggcct ccgcgccggg ttttggcgcc
     4501 tcccgcgggc gcccccctcc tcacggcgag cgctgccacg tcagacgaag ggcgcagcga
     4561 gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc
     4621 cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc
     4681 actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc
     4741 tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt
     4801 ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg
     4861 tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg
     4921 ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga
     4981 gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag
     5041 tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca
     5101 tggtgggcgg caagaaccca aggtcttgag gccttcgcta atgcgggaaa gctcttattc
     5161 gggtgagatg ggctggggca ccatctgggg accctgacgt gaagtttgtc actgactgga
     5221 gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca
     5281 cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt
     5341 ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga
     5401 cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag
     5461 gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt
     5521 agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt
     5581 tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagacta
     5641 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacgga tccataactt
     5701 cgtataatgt atgctatacg aagttatgcc accatgtcta gactggacaa gagcaaagtc
     5761 ataaacggag ctctggaatt actcaatggt gtcggtatcg aaggcctgac gacaaggaaa
     5821 ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg
     5881 gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc
     5941 ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcata ccgctgtgct
     6001 ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag
     6061 tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac
     6121 gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag
     6181 gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt
     6241 ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc
     6301 ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccgacc
     6361 gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac
     6421 cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa
     6481 ataacttcgt atagcataca ttatacgaag ttatggcgcg ccaattccgc ccctctccct
     6541 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     6601 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     6661 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     6721 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     6781 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     6841 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     6901 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     6961 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     7021 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     7081 tcctttgaaa aacacgataa taccatgtat cgatccacca tggtgagcaa gggcgaggag
     7141 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     7201 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     7261 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     7321 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     7381 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     7441 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     7501 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     7561 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     7621 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     7681 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     7741 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     7801 gccgggatca ctctcggcat ggacgagctg tacaagtaat taattaagcg cgtctggaac
     7861 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     7921 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     7981 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     8041 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     8101 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     8161 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     8221 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     8281 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     8341 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     8401 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattaat
     8461 tctgcagtcg agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca
     8521 atgctgattg tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac
     8581 ctcaggtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     8641 aagaaaagag gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     8701 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     8761 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     8821 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     8881 tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat
     8941 gaatatcaga gagtgagagg ccttgacatt gtttaaaccc gctgatcagc ctcgactgtg
     9001 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     9061 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     9121 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     9181 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     9241 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     9301 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     9361 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     9421 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     9481 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     9541 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     9601 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     9661 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     9721 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     9781 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     9841 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     9901 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     9961 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
    10021 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
    10081 acgtgatgaa aaagcctgaa ctcaccgcga cgtgaccgca aggaatcggt caatacacta
    10141 catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga
    10201 tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg
    10261 aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga
    10321 cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc
    10381 aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga
    10441 cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc cgggcgtata
    10501 tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg
    10561 cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc
    10621 gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta gaagtactcg
    10681 ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag cacgtgctac
    10741 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
    10801 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
    10861 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
    10921 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
    10981 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
    11041 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
    11101 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
    11161 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
    11221 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
    11281 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
    11341 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
    11401 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
    11461 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
    11521 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
    11581 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
    11641 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
    11701 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
    11761 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
    11821 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
    11881 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
    11941 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
    12001 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
    12061 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
    12121 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
    12181 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
    12241 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
    12301 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
    12361 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
    12421 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
    12481 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
    12541 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
    12601 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
    12661 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
    12721 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
    12781 gaccgagttg ctcttgcccg gcgtccatac gggataatac cgcgccacat agcagaactt
    12841 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
    12901 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
    12961 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
    13021 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
    13081 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
    13141 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
    13201 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
    13261 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    13321 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
    13381 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
    13441 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
    13501 gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta
    13561 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata
    13621 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga
    13681 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga
    13741 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc
    13801 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact
    13861 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct
    13921 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga
    13981 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
    14041 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
    14101 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tgccgcccga acagggactt
    14161 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
    14221 cacggcaaga ggcgaagggc ggcgactggt gagtacgcca aaaattttga ctagcggaag
    14281 gctagaagga agaagatggt gccagagcgt cagtattaag cgggggaaaa ttagatcgcg
    14341 aagggaaaaa attcggttaa ggccagg
//
