LOCUS       sequence-497733-       12664 bp    DNA     circular SYN 16-JAN-2026
DEFINITION  Constitutively expresses 3xFLAG tagged DR18 with an IgE signal 
            peptide, mNeonGreen and a puromycin resistance gene in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12664)
  AUTHORS   Deol S, Donahue PS, Mitrut RE, Hammitt-Kess IJ, Ahn J, Zhang B, 
            Leonard JN
  TITLE     Comparative Evaluation of Synthetic Cytokines for Enhancing 
            Production and Performance of NK92 Cell-Based Therapies.
  JOURNAL   GEN Biotechnol. 2023 Jun 1;2(3):228-246. doi: 
            10.1089/genbio.2023.0024. Epub 2023 Jun 19.
  PUBMED    37363412
REFERENCE   2  (bases 1 to 12664)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 16, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1089/genbio.2023.0024"; journalName: "GEN Biotechnol"; date: 
            "2023-06-1- 1"; volume: "2"; issue: "3"; pages: "228-246"
COMMENT     Sequence Label: pGIPZ EF1a spIgE-3xFLAG-DR18 IRES
            mNeonGreen-P...
FEATURES             Location/Qualifiers
     source          1..12664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2063..2180
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(2245..2619)
                     /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      2616..2625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2629..2647)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2639..2686)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        2865..4047
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3096..4038
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3995..4015
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     4049..4073
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4148..4157
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4211..4276
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(4775..5349,5350..5352,5353..5361)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4942..4959)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5169..5188
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      5629..5638
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6082..6138
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6139..6741
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MATEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGSRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     6638..6658
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6759..7347
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6812..6832)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7230..7241)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7434..7667
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7768..7785)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7774..7998
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8044..8472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8131..8150)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8341..8362
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8481..8501)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8486..8815
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8666..8801
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8728..8747
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8864..9889
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVHADSGNRRPSTRPRAK
                     E"
     polyA_signal    10019..10140
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10189..10205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10189..10205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10202..10224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10213..10229
                     /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(10237..10243,10244..10261,10262..10267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10282..10303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10420..10437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10591..11179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10671..10690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11350..12141,12142..12210))
                     /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     11973..11992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12211..12315)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12363..12497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     2041 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     2101 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     2161 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttag
     2221 taccgggccc gctctagtcc ggaatcagtc ctgctcctcg gccacgaagt gcacgcagtt
     2281 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     2341 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     2401 ctcgtccagg ccgcgcaccc acacccaggc cagggtgttg tccggcacca cctggtcctg
     2461 gaccgcgctg atgaacaggg tcacgtcgtc ccggaccaca ccggcgaagt cgtcctccac
     2521 gaagtcccgg gagaacccga gccggtcggt ccagaactcg accgctccgg cgacgtcgcg
     2581 cgcggtgagc accggaacgg cactggtcaa cttggccatg gtggccctcc tatagtgagt
     2641 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgcaggtc
     2701 cgaggttcta gagagtaatt catacaaaag gactcgcccc tgccttgggg aatcccaggg
     2761 accgtcgtta aactcccact aacgtagaac ccagagatcg ctgcgttccc gccccctcac
     2821 ccgcccgctc tcgtcatcac tgaggtggag aagagcatgc gtgaggctcc ggtgcccgtc
     2881 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     2941 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     3001 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     3061 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
     3121 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acgcccctgg
     3181 ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg
     3241 ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggct tgggcgctgg
     3301 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     3361 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     3421 gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     3481 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     3541 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     3601 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     3661 gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg
     3721 ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc
     3781 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc
     3841 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     3901 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     3961 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     4021 ttttttcttc catttcaggt gtcgtgagag ctcgtttagt gaaccgtcag atcgcctgga
     4081 gacgccatcc acgctgtttt gacctccata gaagagaacg ggaccgatcc aaattctcga
     4141 ggctagcgcc accatggact ggacctggat tctgtttctg gtggccgctg ccacaagagt
     4201 gcatagcatg gactacaagg accacgacgg cgattacaaa gatcacgaca tcgactataa
     4261 ggacgacgat gacaaaggca gcggctactt cggcaagctg gaaagcaagc tgagcgtgat
     4321 ccggaacctg aacgaccagg tgctgttcat cgaccagggc aacagacccc tgttcgagga
     4381 catgaccgac agcgactgta gagacaacgc ccctcggacc atcttcatca tcagcaagta
     4441 tagcgacagc ctggctagag gcctggccgt gacaatctct gtgaagtgcg agaagatcag
     4501 caccctgagc tgcgagaaca agatcatcag cttcaaagag atgaacccgc cggacaacat
     4561 caaggacacc aagagcgaca tcatcttttt ccagcgggat gtgcccggcc acagccggaa
     4621 gatgcagttt gagagcagca gctacgaggg ctacttcctg gcctgcgaga aagagcggga
     4681 cctgttcaag ctgatcctga agaaagagga cgaactgggc gaccgcagca tcatgttcac
     4741 cgtgcagaac gaggactgag cggccgcgga tccgcccctc tccctccccc ccccctaacg
     4801 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     4861 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     4921 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     4981 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     5041 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     5101 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     5161 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     5221 cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga
     5281 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     5341 gatgataata tggccacaac catgggaact agtatggttt ccaagggcga agaggacaat
     5401 atggccagcc tgcctgccac acacgagctg cacatctttg gcagcatcaa cggcgtggac
     5461 ttcgacatgg tcggacaagg caccggcaat cccaacgacg gctacgagga actgaacctg
     5521 aagtccacaa agggcgacct ccagttcagc ccctggattc tggtgcctca catcggctac
     5581 ggcttccacc agtacctgcc ttatcctgac ggcatgagcc ctttccaggc cgccatggtt
     5641 gatggctctg gctaccaggt ccaccggacc atgcagtttg aggatggcgc ttccctgacc
     5701 gtgaactaca gatacaccta cgagggcagc cacatcaaag gcgaggccca agtgaaaggc
     5761 acaggcttcc ctgctgatgg ccccgtgatg accaattctc tgacagccgc cgactggtgc
     5821 agaagcaaga aaacataccc caacgacaag accatcatca gcaccttcaa gtggtcctac
     5881 accaccggca acggcaagcg gtacagaagc accgccagaa ccacctacac cttcgccaaa
     5941 cctatggccg ccaactacct gaagaaccag cctatgtacg tgttccgcaa gaccgagctg
     6001 aagcactcca agacagaact gaacttcaaa gagtggcaga aagccttcac cgacgtgatg
     6061 ggcatggacg aactctacaa agccacgaac ttcagcctgc tgaagcaggc tggagacgtg
     6121 gaggagaacc ctggacccat ggccaccgag tacaagccca cggtgcgcct cgccacccgc
     6181 gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     6241 cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     6301 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     6361 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggctcg
     6421 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     6481 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     6541 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     6601 gctggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     6661 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     6721 acccgcaagc ccggtgcctg atgtacaagt aggtcgacaa tcaacctctg gattacaaaa
     6781 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     6841 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     6901 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     6961 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     7021 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     7081 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     7141 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     7201 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     7261 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     7321 ggatctccct ttgggccgcc tccccgcctg gaattcgagc tcggtacctt taagaccaat
     7381 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg
     7441 gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt
     7501 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     7561 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     7621 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg
     7681 tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc
     7741 ttgacattgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     7801 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7861 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7921 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     7981 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     8041 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     8101 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     8161 tcacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     8221 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     8281 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     8341 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     8401 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     8461 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     8521 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     8581 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     8641 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     8701 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     8761 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     8821 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgatgaaaa agcctgaact
     8881 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     8941 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     9001 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     9061 ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag
     9121 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     9181 cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga
     9241 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     9301 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     9361 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     9421 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     9481 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     9541 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     9601 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat
     9661 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     9721 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     9781 tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtacacgc
     9841 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtgctacg
     9901 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     9961 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
    10021 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
    10081 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
    10141 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
    10201 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
    10261 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
    10321 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
    10381 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
    10441 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
    10501 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
    10561 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
    10621 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
    10681 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
    10741 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
    10801 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
    10861 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
    10921 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
    10981 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
    11041 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
    11101 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
    11161 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
    11221 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
    11281 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
    11341 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
    11401 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
    11461 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
    11521 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
    11581 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
    11641 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
    11701 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
    11761 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
    11821 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
    11881 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
    11941 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
    12001 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
    12061 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
    12121 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
    12181 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
    12241 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
    12301 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
    12361 tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
    12421 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
    12481 cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca
    12541 ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt
    12601 cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag
    12661 tagt
//
