LOCUS       sequence-497724-       14263 bp    DNA     circular SYN 16-JAN-2026
DEFINITION  Constitutively expresses myc tagged membrane bound IL2, miRFP720 and
            a blasticidin resistance gene in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14263)
  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 14263)
  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 myc tagged mbIL2 IRES miRFP720-P2A-...
FEATURES             Location/Qualifiers
     source          1..14263
                     /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"
     sig_peptide     4154..4213
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             4613..4642
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    join(6317..6891,6892..6894,6895..6903)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6484..6501)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6711..6730
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             7873..7929
                     /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             7942..8340
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8358..8946
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8411..8431)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8829..8840)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9033..9266
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(9367..9384)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9373..9597
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9643..10071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9730..9749)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9940..9961
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10080..10100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10085..10414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10265..10400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             10463..11488
                     /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    11618..11739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11788..11804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11788..11804)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11801..11823)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11812..11828
                     /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(11836..11842,11843..11860,11861..11866))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11881..11902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12019..12036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12190..12778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12270..12289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12949..13740,13741..13809))
                     /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     13572..13591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13810..13914)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13962..14096
                     /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 accatgtacc ggatgcagct gctgtcttgt atcgccctgt ctctggccct
     4201 ggtcaccaat agcgccccta ccagcagcag caccaagaaa acacagctgc aactggaaca
     4261 tctgctgctc gacctccaga tgatcctgaa cggcatcaac aactacaaga accccaagct
     4321 gacccggatg ctgaccttca agttctacat gcccaagaag gccaccgagc tgaagcacct
     4381 ccagtgcctg gaagaggaac tgaagcccct ggaagaagtg ctgaatctgg cccagagcaa
     4441 gaacttccac ctgaggccaa gggacctgat cagcaacatc aacgtgatcg tgctggaact
     4501 gaaaggcagc gagacaacct tcatgtgcga gtacgccgac gagacagcta ccatcgtgga
     4561 atttctgaat cggtggatca ccttctgcca gagcatcatc agcaccctga ccgagcagaa
     4621 gctgatctcc gaagaggacc tggaaatgga aacctctcag ttccccggcg aggaaaagcc
     4681 tcaggcctct cctgaaggca gacccgagag cgaaacaagc tgtgccgtga atggcaccag
     4741 ccagttcacc tgtttctaca acagcaccgc caacatcagc tgcgtgtggt cacaggatgg
     4801 cgctctccag gataccagct gtcaggttca cgcctggcct gatcggagaa gatggaacca
     4861 gacatgcgag ctgctgcccg tgtctcaggc aagctgggcc tgtaatctga tcctgggagc
     4921 ccctgacagc cagaaactga ccaccgtgga tatcgtgacc ctgcgggtgc tctgtagaga
     4981 aggcgttcgt tggagagtga tggccattca ggacttcaag cccttcgaga acctgcggct
     5041 gatggcccca atcagcctgc aagtggtgca tgtggaaacc cacagatgca atatcagctg
     5101 ggagattagc caggccagcc actacttcga gcggcacctg gaatttgagg cccggacact
     5161 gtctccaggc cacacctggg aagaagcccc tctgctgacc ctgaagcaga aacaagagtg
     5221 gatctgcctg gaaaccctga cacctgacac acagtacgag ttccaagtgc gcgtgaagcc
     5281 tctccagggc gagtttacaa catggtcccc ttggagccag cctctggcct ttagaacaaa
     5341 gcctgccgct ctgggcaaag acacaatccc ttggctgggc catctgctcg ttggactgtc
     5401 tggcgctttc ggcttcatca tcctggtgta cctgctgatc aactgccgga acacaggccc
     5461 ctggctgaaa aaggtgctga agtgcaacac ccctgatcct agcaagttct tcagccagct
     5521 gagcagcgag cacggcggag atgttcagaa gtggctgtct agcccatttc ctagcagcag
     5581 cttcagccct ggtggactgg cccctgagat ttccccactg gaagtgctgg aaagggacaa
     5641 agtgacccag ctgctcctcc agcaggataa ggtgccagaa cctgccagcc tgtccagcaa
     5701 tcacagcctg accagctgct ttaccaacca gggctacttc ttcttccatc tgcctgacgc
     5761 actggaaatc gaggcctgcc aggtgtactt cacctacgat ccctacagcg aggaagatcc
     5821 cgacgaaggc gttgcaggcg ctcctacagg atctagccct cagcctctgc aacctctgag
     5881 cggagaggat gacgcctact gcacctttcc aagcagggac gacctgctgc tgttcagccc
     5941 atctctgctc ggaggacctt ctccaccttc tacagcacct ggcggatctg gcgctggcga
     6001 agaaagaatg cctcctagtc tgcaagagcg ggtgcccaga gattgggacc ctcaacctct
     6061 tggccctcca acacctggtg tccccgacct ggtggatttt cagcctcctc ctgaactggt
     6121 gctgagagaa gccggcgaag aggtcccaga tgccggacct agagagggcg ttagcttccc
     6181 ttggtccaga cctcctggcc aaggggagtt tagagccctg aacgctagac tgcccctgaa
     6241 caccgatgcc tacctgtcac tgcaagaact gcaaggacag gaccccacac acctggtcta
     6301 agcggccgcg gatccgcccc tctccctccc ccccccctaa cgttactggc cgaagccgct
     6361 tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg
     6421 gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt
     6481 cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg
     6541 aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac
     6601 ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg
     6661 cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct
     6721 caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg
     6781 atctggggcc tcggtacaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag
     6841 gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa tatggccaca
     6901 accatgggaa ctagtatggc cgagggctct gttgctagac aacccgatct gctgacctgc
     6961 gacgacgagc ccattcatat ccctggcgct atccagcctc acggactgct gcttgctctg
     7021 gccgccgata tgacaatcgt ggccggcagc gataacctgc ctgagctgac aggactggcc
     7081 atcggagccc tgattggaag atcagccgcc gacgtgttcg acagcgagac acacaacaga
     7141 ctgacaatcg ccctggccga acctggtgct gctgtgggag cacctatcac cgtgggcttc
     7201 accatgagaa aggacgccgg ctttatcggc agctggcaca gacacgacca gctgatcttt
     7261 ctggaactgg aacctcctca gagagatgtg gccgagcctc aggccttctt cagaaggacc
     7321 aacagcgcca tcagacggct ccaggccgct gagacactgg aatctgcttg tgccgccgct
     7381 gctcaagaag tgcggaagat caccggcttc gaccgcgtga tgatctacag attcgccagc
     7441 gacttcagcg gcgaagtgat cgccgaggat agatgcgccg aggtggaatc taagctgggc
     7501 ctgcattacc ccgccagctt cattcctgct caggccagac ggctgtatac catcaatccc
     7561 gtgcggatca tccccgacat caactacaga cccgtgcctg tgactcccga tctgaaccct
     7621 gttaccggca gacccatcga cctgagcttc gccatcctga gatccgtgtc tcccgtgcat
     7681 cttgagttca tgcggaacat cggaatgcac ggcaccatga gcatctctat cctgcggggc
     7741 gaaagactgt ggggcctgat cgtgtgtcac cacagaaccc cttactacgt ggacctggat
     7801 ggcagacagg cctgcaaaag agtggccgag agactggcca cacagatcgg cgtgatggaa
     7861 gaaggtagtg gagcaactaa cttttccctg ctcaaacaag ccggggatgt agaggagaac
     7921 cccggccccg ccccccttaa gatggccaag cctttgtctc aagaagaatc caccctcatt
     7981 gaaagagcaa cggctacaat caacagcatc cccatctctg aggactacag cgtcgccagc
     8041 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     8101 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     8161 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     8221 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     8281 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
     8341 tgtacaagta ggtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     8401 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     8461 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     8521 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     8581 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     8641 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     8701 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt
     8761 cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     8821 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     8881 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     8941 ccccgcctgg aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga
     9001 tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag
     9061 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
     9121 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     9181 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     9241 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
     9301 ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgtt taaacccgct
     9361 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     9421 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     9481 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     9541 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     9601 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     9661 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     9721 tagcgcccgc tcctttcgct ttcttccctt cctttctcgt cacgttcgcc ggctttcccc
     9781 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     9841 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     9901 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     9961 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
    10021 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
    10081 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    10141 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
    10201 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
    10261 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
    10321 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
    10381 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
    10441 tttcggatct gatcagcacg tgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa
    10501 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
    10561 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
    10621 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
    10681 gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg
    10741 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
    10801 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
    10861 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
    10921 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
    10981 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
    11041 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
    11101 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
    11161 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
    11221 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
    11281 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
    11341 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
    11401 cgcggccgtc tggaccgatg gctgtgtaga agtacacgcc gatagtggaa accgacgccc
    11461 cagcactcgt ccgagggcaa aggaatagca cgtgctacga gatttcgatt ccaccgccgc
    11521 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
    11581 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
    11641 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
    11701 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
    11761 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
    11821 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
    11881 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
    11941 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
    12001 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
    12061 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
    12121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
    12181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
    12241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
    12301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
    12361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
    12421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
    12481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
    12541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
    12601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
    12661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
    12721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
    12781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
    12841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
    12901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
    12961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
    13021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
    13081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
    13141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
    13201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
    13261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
    13321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
    13381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
    13441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
    13501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
    13561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
    13621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
    13681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
    13741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
    13801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
    13861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
    13921 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt
    13981 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
    14041 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact
    14101 ggataactca agctaaccaa aatcatccca aacttcccac cccataccct attaccactg
    14161 ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt
    14221 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
