LOCUS       sequence-491540-       12464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of an ER-specific SUNTAG reporter; piggyBac 
            vector to express cytERM-SUNTAG MS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12464)
  AUTHORS   Choi H, Liao YC, Yoon YJ, Grimm J, Wang N, Lavis LD, Singer RH, 
            Lippincott-Schwartz J
  TITLE     Secretome translation shaped by lysosomes and lunapark-marked ER 
            junctions.
  JOURNAL   Nature. 2025 Nov 5. doi: 10.1038/s41586-025-09718-0.
  PUBMED    41193816
REFERENCE   2  (bases 1 to 12464)
  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: "Nature. 
            2025 Nov 5. doi: 10.1038/s41586-025-09718-0."
COMMENT     Sequence Label: pPB-cytERM-SUNTAG-MS2
FEATURES             Location/Qualifiers
     source          1..12464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1474..1515
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1532..1549
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1606..1623)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1612..1819
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1896..2107
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2120..2388
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2422..3165
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             3166..3219
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3220..3819
                     /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(3220..3239)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3716..3736
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3858..3979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3895..3914)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3949..3968
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4741..4775
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(5266..5282)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5266..5282)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5279..5301)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5290..5306
                     /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(5314..5320,5321..5338,5339..5344))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5359..5380
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5497..5514)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5668..6256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5748..5767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6427..7218,7219..7287))
                     /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     7050..7069
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7288..7392)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7418..7873)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7555..7576)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7767..7786
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8000..8022
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8014..8031
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8015..8031
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   8161..8223
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    8676..8694
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8712..8730
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8748..8766
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8784..8802
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8820..8838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8856..8874
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    8892..8910
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     8955..8979
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             9227..9925
                     /codon_start=1
                     /product="far-red monomeric fluorescent protein with an 
                     emission peak at 657 nm (Morozova et al., 2010)"
                     /label=TagRFP657
                     /note="derived from mKate"
                     /translation="MSELITENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTQRIKV
                     VEGGPLPFAFDILATSFMYGSHTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEAHTEMLYPADGGLEGRTALALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDYRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             9965..10021
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10037..10093
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10109..10165
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10181..10237
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10253..10309
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10325..10381
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10397..10453
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10469..10525
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10541..10597
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10613..10669
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10685..10741
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10757..10813
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10829..10885
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10901..10957
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             10973..11029
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             11045..11101
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             11117..11173
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             11189..11245
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             11261..11317
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             11333..11389
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             11405..11461
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             11477..11533
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             11621..11677
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             join(11729..11731,11732..11734,11735..12442)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLSHGVQVFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELY"
     primer_bind     complement(11774..11795)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 ggttttgtct ttttttaata gtcattccaa gtatccatga aataagtggt tacaggaagt
       61 ccctcaccct cccaaaagcc acccccactc ctaagaggag gatggtcgcg tccatgccct
      121 gagtccaccc cggggaaggt gacaaccggt aacctacaaa cgggtggagg atcaccccac
      181 ccgacacttc acaatcaagg ggtacaatac acaagggtgg aggaacaccc caccctccag
      241 acacattaca cagaaatcca atcaaacaga agcaccatca gggcttctgc taccaaattt
      301 atctcaaaaa actacaacaa ggaatcacca tcagggattc cctgtgcaat atacgtcaaa
      361 cgagggccac gacgggagga cgatcacgcc tcccgaatat cggcatgtct ggctttcgaa
      421 ttcagtgcgt ggagcatcag cccacgcagc caatcagagt cgaatacaag tcgactttcg
      481 cgaagagcat cagccttcgc gccattctta cacaaaccac actctcccct acaggaacag
      541 catcagcgtt cctgcccagt acccaactca agaaaattta tgtccccatg cagcatcagc
      601 gcatgggccc caagaataca tccccaacaa aatcacatcc gagcaccaac agggctagga
      661 gtgttgtttc ttgtccaact ggacaaaccc tccatggacc atcaggccat ggactctcac
      721 caacaagaca aaaactactc ttctcgaagc agcatcagcg cttcgaaaca ctcgagcata
      781 cattgtgcct atttcttggg tggacgatca cgccacccat gctctcacga atttcaaaac
      841 acggacaagg acgagcacca ccagggctcg tcgttccacg tccaatacga ttacttacct
      901 ttcgggatca cgatcacgga tcccgcagct acatcacttc cactcaggac attcaagcat
      961 gcacgatcac ggcatgctcc acaagtctca accacagaaa ctaccaaatg ggttcagcac
     1021 cagcgaaccc actcctacct caaacctctt cccacaaaac tggcaagcag gatcaccgct
     1081 tgcccattcc aacataccaa atcaaaaaca attactggta cagcatcagc gtaccagccc
     1141 acatctctca ctactatcaa aaaccaaacc gttcagcaac agcgaacggt acacacggaa
     1201 aaatcaactg gtttacaaat acgaaagacg atcacgcttt cgtccagcgc aaactattac
     1261 gaaaaacatc cgacgggaag agcaacagcc ttcccgcggc ggaaaacctc acaaaaacac
     1321 gacaaacgga tgcacgaaca cggcatccgc cgacaaccca caaacttaca accaggcaaa
     1381 cggtgcagga tcaccgcacc gtacatcaaa cacctcagat ctcatatcga tagatcctaa
     1441 tcaacctctg gattacaaat gtacaagctc gagggtaagc ctatccctaa ccctctcctc
     1501 ggtctcgatt ctacgtagta atgaaccggt ccaccaccac caccaccact aaggatccgg
     1561 ggttggggtt gcgccttttc caagcacctg ctacatagcg gtacccctcg actgtgcctt
     1621 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1681 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1741 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1801 atagcaggca tgctggggat gcggtgggct cttcgaattt aaatcggatc cagctgaatc
     1861 taagtggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     1921 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     1981 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     2041 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     2101 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     2161 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     2221 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     2281 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     2341 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcta agctgtgacc
     2401 ggcgcctact ctagagccgc catgtctaga ctggacaaga gcaaagtcat aaactctgct
     2461 ctggaattac tcaatggagt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag
     2521 ctgggagttg agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat
     2581 gccctgccaa tcgagatgct ggacaggcat catacccact cctgccccct ggaaggcgag
     2641 tcatggcaag actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat
     2701 cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg
     2761 gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct
     2821 ctgtccgccg tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta
     2881 gcaaaagagg aaagagagac acctaccacc gattctatgc ccccacttct gaaacaagca
     2941 attgagctgt tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc
     3001 atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac
     3061 gattttgact tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg
     3121 cctgctgacg ctcttgacga ttttgacctt gacatgctcc ccggggaggg cagaggaagt
     3181 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg
     3241 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     3301 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     3361 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     3421 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     3481 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     3541 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     3601 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     3661 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     3721 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     3781 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgag tcgacaatca acctctggat
     3841 tacaaaattt gtgggttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3901 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3961 aactcatcaa tgtatcttat catgtctgga attgactcaa atgatgtcaa ttagtctatc
     4021 agaagctatc tggtctccct tccgggggac aagacatccc tgtttaatat ttaaacagca
     4081 gtgttcccaa actgggttct tatatccctt gctctggtca accaggttgc agggtttcct
     4141 gtcctcacag gaacgaagtc cctaaagaaa cagtggcagc caggtttagc cccggaattg
     4201 actggattcc ttttttaggg cccattggta tggctttttc cccgtatccc cccaggtgtc
     4261 tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc caccttcccc
     4321 gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg ccggaccgga
     4381 gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa ttacatccct
     4441 gggggctttg ggggggggct gtccctgata tctataacaa gaaaatatat atataataag
     4501 ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa
     4561 gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca
     4621 gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt
     4681 cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg
     4741 catgcgtcaa ttttacgcag actatctttc tagggttaat ctagctgcat caggatcata
     4801 tcgtcgggtc ttttttccgg ctcagtcatc gcccaagctg gcgctatctg ggcatcgggg
     4861 aggaagaagc ccgtgccttt tcccgcgagg ttgaagcggc atggaaagag tttgccgagg
     4921 atgactgctg ctgcattgac gttgagcgaa aacgcacgtt taccatgatg attcgggaag
     4981 gtgtggccat gcacgccttt aacggtgaac tgttcgttca ggccacctgg gataccagtt
     5041 cgtcgcggct tttccggaca cagttccgga tggtcagccc gaagcgcatc agcaacccga
     5101 acaataccgg cgacagccgg aactgccgtg ccggtgtgca gattaatgac agcggtgcgg
     5161 cgctgggata ttacgtcagc gaggacgggt atcctggctg gatgccgcag aaatggacat
     5221 ggataccccg tgagttaccc ggcgggcgcg cttggcgtaa tcatggtcat agctgtttcc
     5281 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5341 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5401 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5461 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5521 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5581 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5641 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5701 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5761 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5821 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5881 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5941 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6001 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6061 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6121 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6181 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6241 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6301 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6361 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6421 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6481 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6541 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6601 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6661 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6721 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6781 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6841 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6901 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6961 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7021 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7081 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7141 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7201 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7261 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7321 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7381 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     7441 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     7501 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     7561 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     7621 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     7681 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     7741 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     7801 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     7861 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7921 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     7981 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcc
     8041 tcgttcattc acgtttttga acccgtggag gacgggcaga ctcgcggtgc aaatgtgttt
     8101 tacagcgtga tggagcagat gaagatgctc gacacgctgc agaacacgca gctagattaa
     8161 ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgtgta aaattgacgc
     8221 atgtgtttta tcggtctgta tatcgaggtt tatttattaa tttgaataga tattaagttt
     8281 tattatattt acacttacat actaataata aattcaacaa acaatttatt tatgtttatt
     8341 tatttattaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa acttttatga
     8401 gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc gctagggggc
     8461 agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc cccgagccgg
     8521 cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt cctctgaacg
     8581 cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa ggcctccacg
     8641 gccactagtc tttcgtcttc aagaattccg tttactccct atcagtgata gagaacgtat
     8701 gaagagttta ctccctatca gtgatagaga acgtatgcag actttactcc ctatcagtga
     8761 tagagaacgt ataaggagtt tactccctat cagtgataga gaacgtatga ccagtttact
     8821 ccctatcagt gatagagaac gtatctacag tttactccct atcagtgata gagaacgtat
     8881 atccagttta ctccctatca gtgatagaga acgtataagc tttaggcgtg tacggtgggc
     8941 gcctataaaa gcagagctcg tttagtgaac cgtcagatcg cctggagcaa ttccacaaca
     9001 cttttgtctt ataccaactt tccgtaccac ttcctaccct cgtaaaggtc tagagctagc
     9061 atcgctgtga tcgtcacttg gcggccgcat ggatcctgtg gtggtgctgg ggctctgtct
     9121 ctcctgtttg cttctccttt cactctggaa acagagctat gggggaggga agcttcgaat
     9181 tctgcagtcg acggtaccgc gggcccggga tccaccggtc gccaccatga gcgagctgat
     9241 caccgagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
     9301 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagacccaga gaatcaaggt
     9361 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
     9421 cagtcacacc ttcatcaacc acacccaggg catccccgac ttctggaagc agtccttccc
     9481 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
     9541 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     9601 cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg cccacaccga
     9661 gatgctgtac cccgctgacg gcggcctgga aggcagaacc gcgctggccc tgaagctcgt
     9721 gggcgggggc cacctgatct gcaacttcaa gaccacatac agatccaaga aacccgctaa
     9781 gaacctcaag atgcccggcg tctactatgt ggactacaga ctggaaagaa tcaaggaggc
     9841 cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc
     9901 tagcaaactg gggcacaaac ttaatggcgg accgggtgga tctggaggtg gaggttctgg
     9961 aggagaagaa cttttgagca agaattatca tcttgagaac gaagtggctc gtcttaagaa
    10021 aggttctggc agtggagaag aactgctttc aaagaattac cacctggaaa atgaggtagc
    10081 tagactgaaa aaggggagcg gaagtgggga ggagttgctg agcaaaaatt atcatttgga
    10141 gaacgaagta gcacgactaa agaaagggtc cggatcgggt gaggagttac tctcgaaaaa
    10201 ttatcatctc gaaaacgaag tggctcggct aaaaaagggc agtggttctg gagaagagct
    10261 attatctaaa aactaccacc tcgaaaatga ggtggcacgc ttaaaaaagg gaagtggcag
    10321 tggtgaagag ctactatcca agaattatca tcttgagaac gaggtagcgc gtttgaagaa
    10381 gggttccggc tcaggagagg aactgctctc gaagaactat catcttgaaa atgaggtcgc
    10441 tcgattaaaa aagggatcgg gcagtggtga ggaactactt tcaaagaatt accacctcga
    10501 aaacgaagta gctcgattaa agaaaggttc agggtcgggt gaagaattac tgagtaaaaa
    10561 ttatcatctg gaaaatgagg tagcgagact aaaaaagggg agtggttctg gcgaagagtt
    10621 gctatcgaaa aattatcatc ttgagaacga agttgctagg ctcaaaaagg gctcaggctc
    10681 aggcgaggag ttgctctcga aaaactacca cttggaaaat gaggtcgcga ggttgaaaaa
    10741 ggggagcggg tcgggcgagg agttattgag caaaaactat catttagaga acgaagtcgc
    10801 gcgcttaaag aaaggctcgg gctcgggcga agaactctta tcgaagaact accacctcga
    10861 aaatgaggtc gccaggttga aaaagggcag tggcagcggg gaggaactct tgagcaagaa
    10921 ctaccacttg gagaatgagg tcgcgagatt gaagaaaggg tcggggagcg gcgaggaatt
    10981 gctcagcaag aattatcatt tggagaacga agtcgccagg ctcaagaaag gctcggggtc
    11041 gggggaggaa ttgttgagta aaaactacca cttggaaaat gaagtcgcca ggctcaaaaa
    11101 agggagtggg agcggcgaag agttattgag caaaaattac cacttggaga acgaagtggc
    11161 aaggctcaag aaagggagcg gcagcgggga ggagctctta tcgaagaact accacttaga
    11221 gaatgaagtc gcccgcttga agaaaggctc ggggagcggg gaagagctct tgagcaagaa
    11281 ctaccacttg gaaaatgagg tggcgcgctt gaagaaaggg agcgggagcg gggaagagtt
    11341 actatctaag aattatcatc tcgagaacga ggtggctcga ctaaagaagg gctccggcag
    11401 tggggaggaa ctcctgtcga agaactatca tcttgaaaat gaggttgcaa gacttaaaaa
    11461 ggggtccgga tcaggtgagg aactactcag taagaattac cacctggaaa acgaagttgc
    11521 acgtttgaag aaaggatcag gatcaggcga agaactgctc tcaaaagatt atcatttgga
    11581 aaatgaggtt gcacgtttaa aaaagggaag tggcagtggt gaggaacttc tgtcgaaaaa
    11641 ttatcatctc gagaatgaag tagcccgact taaaaagggt tctggctcgg gtggaggcgg
    11701 tggatcaggt ggaggaggtt cactgcagat ggtgagcaag ggcgaggagc tgttcaccgg
    11761 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgag
    11821 gggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca tcagtactac
    11881 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgagccacg gcgtgcaggt
    11941 gttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
    12001 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcacctaca agacccgcgc
    12061 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcgtcgactt
    12121 caaggaggac ggcaacatcc tggggcacaa gctggagtac aacttcaaca gccacaacat
    12181 ctatatcatg gccgtcaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa
    12241 cgtggaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
    12301 cggccccgtg ctgctgcccg acagccacta cctgagcacc cagtccgtgc tgagcaaaga
    12361 ccccaacgag aagcgcgatc acatggtcct gctggagttc cgcaccgccg ccgggatcac
    12421 tctcggcatg gacgagctgt acattgtttt gttttttttt tttt
//
