LOCUS       sequence-497728-       12496 bp    DNA     circular SYN 16-JAN-2026
DEFINITION  Constitutively expresses 3xFLAG tagged Neoleukin 2/15 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 12496)
  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 12496)
  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-Neo2/15 IRES
            mNeonGree...
FEATURES             Location/Qualifiers
     source          1..12496
                     /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(4607..5181,5182..5184,5185..5193)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4774..4791)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5001..5020
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      5461..5470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5914..5970
                     /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             5971..6573
                     /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     6470..6490
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6591..7179
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6644..6664)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7062..7073)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7266..7499
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7600..7617)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7606..7830
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7876..8304
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7963..7982)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8173..8194
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8313..8333)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8318..8647
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8498..8633
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8560..8579
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8696..9721
                     /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    9851..9972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10021..10037)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10021..10037)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10034..10056)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10045..10061
                     /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(10069..10075,10076..10093,10094..10099))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10114..10135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10252..10269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10423..11011)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10503..10522)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11182..11973,11974..12042))
                     /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     11805..11824
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12043..12147)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12195..12329
                     /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 gcggcatgcc caagaagaag atccagctgc acgccgagca
     4321 cgccctgtat gatgccctga tgatcctgaa catcgtcaag acaaacagcc ctcctgccga
     4381 ggaaaagctg gaagattacg ccttcaactt cgagctgatc ctggaagaga tcgcccggct
     4441 gtttgagagc ggcgaccaga aggatgaggc cgagaaggcc aagcggatga aggaatggat
     4501 gaagcggatc aagaccaccg ccagcgagga cgagcaagag gaaatggcca acgccatcat
     4561 caccatcctc cagtcctgga tctttagcta agcggccgcg gatccgcccc tctccctccc
     4621 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     4681 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     4741 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     4801 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     4861 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     4921 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     4981 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     5041 cccagaaggt accccattgt atgggatctg atctggggcc tcggtacaca tgctttacat
     5101 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     5161 tttgaaaaac acgatgataa tatggccaca accatgggaa ctagtatggt ttccaagggc
     5221 gaagaggaca atatggccag cctgcctgcc acacacgagc tgcacatctt tggcagcatc
     5281 aacggcgtgg acttcgacat ggtcggacaa ggcaccggca atcccaacga cggctacgag
     5341 gaactgaacc tgaagtccac aaagggcgac ctccagttca gcccctggat tctggtgcct
     5401 cacatcggct acggcttcca ccagtacctg ccttatcctg acggcatgag ccctttccag
     5461 gccgccatgg ttgatggctc tggctaccag gtccaccgga ccatgcagtt tgaggatggc
     5521 gcttccctga ccgtgaacta cagatacacc tacgagggca gccacatcaa aggcgaggcc
     5581 caagtgaaag gcacaggctt ccctgctgat ggccccgtga tgaccaattc tctgacagcc
     5641 gccgactggt gcagaagcaa gaaaacatac cccaacgaca agaccatcat cagcaccttc
     5701 aagtggtcct acaccaccgg caacggcaag cggtacagaa gcaccgccag aaccacctac
     5761 accttcgcca aacctatggc cgccaactac ctgaagaacc agcctatgta cgtgttccgc
     5821 aagaccgagc tgaagcactc caagacagaa ctgaacttca aagagtggca gaaagccttc
     5881 accgacgtga tgggcatgga cgaactctac aaagccacga acttcagcct gctgaagcag
     5941 gctggagacg tggaggagaa ccctggaccc atggccaccg agtacaagcc cacggtgcgc
     6001 ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac
     6061 taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg
     6121 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     6181 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     6241 gagatcggct cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     6301 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     6361 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     6421 gcggccgagc gcgctggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     6481 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     6541 acctggtgca tgacccgcaa gcccggtgcc tgatgtacaa gtaggtcgac aatcaacctc
     6601 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     6661 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     6721 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     6781 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     6841 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     6901 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     6961 acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg
     7021 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     7081 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     7141 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaattcga gctcggtacc
     7201 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaagag
     7261 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     7321 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     7381 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     7441 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     7501 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     7561 gagtgagagg ccttgacatt gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     7621 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     7681 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     7741 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     7801 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     7861 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     7921 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     7981 cttcctttct cgtcacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     8041 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     8101 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     8161 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     8221 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     8281 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     8341 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     8401 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     8461 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     8521 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     8581 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     8641 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgatgaa
     8701 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     8761 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     8821 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     8881 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     8941 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     9001 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     9061 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     9121 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     9181 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     9241 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     9301 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     9361 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     9421 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     9481 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     9541 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     9601 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     9661 agaagtacac gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     9721 gcacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     9781 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     9841 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     9901 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     9961 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
    10021 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
    10081 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
    10141 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
    10201 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
    10261 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
    10321 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
    10381 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
    10441 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
    10501 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
    10561 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
    10621 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
    10681 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
    10741 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
    10801 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
    10861 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
    10921 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
    10981 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
    11041 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
    11101 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
    11161 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
    11221 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
    11281 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
    11341 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
    11401 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
    11461 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
    11521 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
    11581 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
    11641 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
    11701 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
    11761 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
    11821 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
    11881 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
    11941 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
    12001 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
    12061 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
    12121 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
    12181 cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
    12241 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
    12301 tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac caaaatcatc
    12361 ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt catttactct
    12421 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
    12481 ctacaaactt agtagt
//
