LOCUS       sequence-497729-       13692 bp    DNA     circular SYN 16-JAN-2026
DEFINITION  Constitutively expresses an engineered version of IL12, 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 13692)
  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 13692)
  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 ACCx2 uORF eIL12 IRES mNeonGreen-P2...
FEATURES             Location/Qualifiers
     source          1..13692
                     /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"
     misc_feature    join(5803..6377,6378..6380,6381..6389)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5970..5987)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6197..6216
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      6657..6666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7110..7166
                     /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             7167..7769
                     /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     7666..7686
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7787..8375
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7840..7860)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8258..8269)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8462..8695
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8796..8813)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8802..9026
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9072..9500
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9159..9178)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9369..9390
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9509..9529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9514..9843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9694..9829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9756..9775
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             9892..10917
                     /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    11047..11168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11217..11233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11217..11233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11230..11252)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11241..11257
                     /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(11265..11271,11272..11289,11290..11295))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11310..11331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11448..11465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11619..12207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11699..11718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12378..13169,13170..13238))
                     /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     13001..13020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13239..13343)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13391..13525
                     /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 ggctagcacc atgggttgac caccatgtgt caccagcagc tggtcatcag ctggttcagc
     4201 ctggtgttcc tggcctctcc tctggtggcc atctgggagc tgaagaaaga cgtgtacgtg
     4261 gtggaactgg actggtatcc cgatgctcct ggcgagatgg tggtgctgac ctgcgatacc
     4321 cctgaagagg acggcatcac ctggacactg gatcagtcta gcgaggtgct cggcagcggc
     4381 aagaccctga ccatccaagt gaaagagttt ggcgacgccg gccagtacac ctgtcacaaa
     4441 ggcggagaag tgctgagcca cagcctgctg ctgctccaca agaaagagga tggcatttgg
     4501 agcaccgaca tcctgaagga ccagaaagag cccaagaaca agaccttcct gagatgcgag
     4561 gccaagaact acagcggccg gttcacatgt tggtggctga ccaccatcag caccgacctg
     4621 accttcagcg tgaagtccag cagaggcagc agtgatcctc agggcgttac atgtggcgcc
     4681 gctacactgt ctgccgaaag agtgcggggc gacaacaaag aatacgagta cagcgtggaa
     4741 tgccaagagg acagcgcctg tccagccgcc gaagagtctc tgcctatcga agtgatggtg
     4801 gacgccgtgc acaagctgaa gtacgagaac tacacctcca gctttttcat ccgggacatc
     4861 atcaagcccg atcctccaaa gaatctccag ctgaagcctc tgaagaacag cagacaggtg
     4921 gaagtgtcct gggagtaccc cgacacctgg tctacacccc acagctactt cagcctgacc
     4981 ttttgcgtgc aagtgcaggg caagtccaag cgcgagaaaa aggaccgggt gttcaccgac
     5041 aagaccagcg ccaccgtgat ctgccggaag aatgccagca tcagcgtcag agcccaggat
     5101 cggtactaca gcagctcttg gagcgagtgg gccagcgttc catgttctgg tggcggaggc
     5161 agcggaggcg gaggcagcgg cggaggaggc agcagaaatc tgcctgttgc cactcctgat
     5221 cctggcatgt tcccttgcct gcaccacagc cagaatctgc tgagagccgt gtccaacatg
     5281 ctccagaagg ccagacagac ccttgagttc tacccctgca ccagcgagga aatcgaccac
     5341 gaggacatca ccaaggataa gaccagcacc gtggaagcct gcctgcctct ggaactgacc
     5401 aagaacgaga gctgcctgaa cagccgggaa accagcttca tcaccaacgg ctcttgcctg
     5461 gccagcagaa agacctcctt catgatggcc ctgtgcctga gcagcatcta cgaggacctg
     5521 aagatgtacc aggtcgagtt caagaccatg aacgccaagc tgctgatgga ccccaagcgg
     5581 cagattttcc tggaccagaa tatgctggcc gtgatcgacg agctgatgca ggccctgaac
     5641 ttcaacagcg agacagtgcc ccagaagtct agcctggaag aacccgactt ctacaagacc
     5701 aagatcaagc tgtgcatcct gctgcacgcc ttcagaatca gagccgtgac catcgacaga
     5761 gtgatgagct acctgaacgc cagctaagcg gccgcggatc cgcccctctc cctccccccc
     5821 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     5881 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     5941 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     6001 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     6061 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     6121 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     6181 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     6241 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt
     6301 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     6361 aaaaacacga tgataatatg gccacaacca tgggaactag tatggtttcc aagggcgaag
     6421 aggacaatat ggccagcctg cctgccacac acgagctgca catctttggc agcatcaacg
     6481 gcgtggactt cgacatggtc ggacaaggca ccggcaatcc caacgacggc tacgaggaac
     6541 tgaacctgaa gtccacaaag ggcgacctcc agttcagccc ctggattctg gtgcctcaca
     6601 tcggctacgg cttccaccag tacctgcctt atcctgacgg catgagccct ttccaggccg
     6661 ccatggttga tggctctggc taccaggtcc accggaccat gcagtttgag gatggcgctt
     6721 ccctgaccgt gaactacaga tacacctacg agggcagcca catcaaaggc gaggcccaag
     6781 tgaaaggcac aggcttccct gctgatggcc ccgtgatgac caattctctg acagccgccg
     6841 actggtgcag aagcaagaaa acatacccca acgacaagac catcatcagc accttcaagt
     6901 ggtcctacac caccggcaac ggcaagcggt acagaagcac cgccagaacc acctacacct
     6961 tcgccaaacc tatggccgcc aactacctga agaaccagcc tatgtacgtg ttccgcaaga
     7021 ccgagctgaa gcactccaag acagaactga acttcaaaga gtggcagaaa gccttcaccg
     7081 acgtgatggg catggacgaa ctctacaaag ccacgaactt cagcctgctg aagcaggctg
     7141 gagacgtgga ggagaaccct ggacccatgg ccaccgagta caagcccacg gtgcgcctcg
     7201 ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc
     7261 ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag
     7321 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     7381 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     7441 tcggctcgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     7501 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     7561 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     7621 ccgagcgcgc tggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     7681 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     7741 ggtgcatgac ccgcaagccc ggtgcctgat gtacaagtag gtcgacaatc aacctctgga
     7801 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     7861 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     7921 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     7981 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     8041 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     8101 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     8161 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     8221 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     8281 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     8341 gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc ggtaccttta
     8401 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagagggga
     8461 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     8521 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     8581 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     8641 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     8701 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     8761 gagaggcctt gacattgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     8821 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     8881 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     8941 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     9001 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     9061 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     9121 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     9181 ctttctcgtc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     9241 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     9301 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     9361 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     9421 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     9481 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     9541 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     9601 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     9661 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     9721 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     9781 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     9841 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gatgaaaaag
     9901 cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc
     9961 gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg
    10021 cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt
    10081 tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc
    10141 agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg
    10201 cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct
    10261 gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa
    10321 tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa
    10381 actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt
    10441 tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat
    10501 gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg
    10561 gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag
    10621 cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg
    10681 gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc
    10741 gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact
    10801 gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa
    10861 gtacacgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac
    10921 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
    10981 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
    11041 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
    11101 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
    11161 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
    11221 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
    11281 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
    11341 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
    11401 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
    11461 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
    11521 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
    11581 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
    11641 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
    11701 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
    11761 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    11821 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    11881 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    11941 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    12001 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
    12061 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    12121 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    12181 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
    12241 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
    12301 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
    12361 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
    12421 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
    12481 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
    12541 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
    12601 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    12661 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
    12721 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
    12781 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
    12841 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    12901 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    12961 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    13021 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    13081 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    13141 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    13201 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    13261 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    13321 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
    13381 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
    13441 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
    13501 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
    13561 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac
    13621 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac
    13681 aaacttagta gt
//
