LOCUS       sequence-497732-       12610 bp    DNA     circular SYN 16-JAN-2026
DEFINITION  Constitutively expresses 3xFLAG tagged DR18, 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 12610)
  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 12610)
  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 spnull-3xFLAG-DR18 IRES
            mNeonGreen-...
FEATURES             Location/Qualifiers
     source          1..12610
                     /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             4157..4222
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    join(4721..5295,5296..5298,5299..5307)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4888..4905)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5115..5134
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      5575..5584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6028..6084
                     /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             6085..6687
                     /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     6584..6604
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6705..7293
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6758..6778)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7176..7187)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7380..7613
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7714..7731)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7720..7944
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7990..8418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8077..8096)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8287..8308
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8427..8447)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8432..8761
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8612..8747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8674..8693
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             8810..9835
                     /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    9965..10086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10135..10151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10135..10151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10148..10170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10159..10175
                     /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(10183..10189,10190..10207,10208..10213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10228..10249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10366..10383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10537..11125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10617..10636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11296..12087,12088..12156))
                     /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     11919..11938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12157..12261)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    12309..12443
                     /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 acaaggacca cgacggcgat tacaaagatc acgacatcga
     4201 ctataaggac gacgatgaca aaggcagcgg ctacttcggc aagctggaaa gcaagctgag
     4261 cgtgatccgg aacctgaacg accaggtgct gttcatcgac cagggcaaca gacccctgtt
     4321 cgaggacatg accgacagcg actgtagaga caacgcccct cggaccatct tcatcatcag
     4381 caagtatagc gacagcctgg ctagaggcct ggccgtgaca atctctgtga agtgcgagaa
     4441 gatcagcacc ctgagctgcg agaacaagat catcagcttc aaagagatga acccgccgga
     4501 caacatcaag gacaccaaga gcgacatcat ctttttccag cgggatgtgc ccggccacag
     4561 ccggaagatg cagtttgaga gcagcagcta cgagggctac ttcctggcct gcgagaaaga
     4621 gcgggacctg ttcaagctga tcctgaagaa agaggacgaa ctgggcgacc gcagcatcat
     4681 gttcaccgtg cagaacgagg actgagcggc cgcggatccg cccctctccc tccccccccc
     4741 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     4801 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     4861 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     4921 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     4981 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     5041 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     5101 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     5161 aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt
     5221 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     5281 aaacacgatg ataatatggc cacaaccatg ggaactagta tggtttccaa gggcgaagag
     5341 gacaatatgg ccagcctgcc tgccacacac gagctgcaca tctttggcag catcaacggc
     5401 gtggacttcg acatggtcgg acaaggcacc ggcaatccca acgacggcta cgaggaactg
     5461 aacctgaagt ccacaaaggg cgacctccag ttcagcccct ggattctggt gcctcacatc
     5521 ggctacggct tccaccagta cctgccttat cctgacggca tgagcccttt ccaggccgcc
     5581 atggttgatg gctctggcta ccaggtccac cggaccatgc agtttgagga tggcgcttcc
     5641 ctgaccgtga actacagata cacctacgag ggcagccaca tcaaaggcga ggcccaagtg
     5701 aaaggcacag gcttccctgc tgatggcccc gtgatgacca attctctgac agccgccgac
     5761 tggtgcagaa gcaagaaaac ataccccaac gacaagacca tcatcagcac cttcaagtgg
     5821 tcctacacca ccggcaacgg caagcggtac agaagcaccg ccagaaccac ctacaccttc
     5881 gccaaaccta tggccgccaa ctacctgaag aaccagccta tgtacgtgtt ccgcaagacc
     5941 gagctgaagc actccaagac agaactgaac ttcaaagagt ggcagaaagc cttcaccgac
     6001 gtgatgggca tggacgaact ctacaaagcc acgaacttca gcctgctgaa gcaggctgga
     6061 gacgtggagg agaaccctgg acccatggcc accgagtaca agcccacggt gcgcctcgcc
     6121 acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     6181 gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     6241 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     6301 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     6361 ggctcgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     6421 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     6481 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     6541 gagcgcgctg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     6601 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     6661 tgcatgaccc gcaagcccgg tgcctgatgt acaagtaggt cgacaatcaa cctctggatt
     6721 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     6781 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     6841 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     6901 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     6961 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     7021 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     7081 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     7141 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     7201 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     7261 cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacctttaag
     7321 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact
     7381 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     7441 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     7501 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     7561 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     7621 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     7681 gaggccttga cattgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     7741 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     7801 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     7861 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     7921 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     7981 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     8041 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     8101 ttctcgtcac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     8161 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     8221 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     8281 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     8341 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     8401 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     8461 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     8521 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     8581 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     8641 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     8701 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     8761 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtga tgaaaaagcc
     8821 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     8881 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     8941 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     9001 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     9061 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     9121 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     9181 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     9241 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     9301 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     9361 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     9421 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     9481 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     9541 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     9601 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     9661 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     9721 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     9781 acacgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aatagcacgt
     9841 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     9901 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     9961 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
    10021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
    10081 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
    10141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
    10201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
    10261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
    10321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    10381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    10441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    10501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    10561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    10621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    10681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    10741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    10801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    10861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    10921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    10981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    11041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    11101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    11161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    11221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    11281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    11341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    11401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    11461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    11521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    11581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    11641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    11701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    11761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    11821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    11881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    11941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    12001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    12061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    12121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    12181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    12241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
    12301 gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
    12361 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
    12421 atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac
    12481 ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct
    12541 gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa
    12601 acttagtagt
//
