LOCUS       sequence-331143-        9794 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible NKX2-1 overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9794)
  AUTHORS   Sun D, Evans L, Perrone F, Sokleva V, Lim K, Rezakhani S, Lutolf M, 
            Zilbauer M, Rawlins EL
  TITLE     A functional genetic toolbox for human tissue-derived organoids.
  JOURNAL   eLife. 2021 Oct 6;10. pii: 67886. doi: 10.7554/eLife.67886.
  PUBMED    34612202
REFERENCE   2  (bases 1 to 9794)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "eLife."; 
            date: "2021-10-6"; volume: "10. pii: 67886. doi"; pages: " 
            10.7554/eLife.67886"
COMMENT     Sequence Label: pLenti-tetON-NKX2-1-EF1a-TagRFP-2A-tet3G
FEATURES             Location/Qualifiers
     source          1..9794
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    2246..2264
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2282..2300
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2318..2336
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2354..2372
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2390..2408
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2426..2444
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2462..2480
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2525..2549
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(3797..3816)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3862..3917
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3916..3935)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        4121..4332
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             4345..4613
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      4648..4657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4654..5364
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     CDS             5371..5424
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5425..6171
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    6184..6772
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6237..6257)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6655..6666)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6847..7080
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7199..7217)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7285..7389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7390..7458,7459..8250)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7608..7627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8421..9009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8910..8929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9163..9180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9420..9441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9456..9461,9462..9479,9480..9486)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9494..9510
                     /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)."
     primer_bind     9499..9521
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9518..9534
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9518..9534
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9584..9603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
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 atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagaccg
     2221 gtggcctcca cggccactag attactccct atcagtgata gagaacgtat gaagagttta
     2281 ctccctatca gtgatagaga acgtatgcag actttactcc ctatcagtga tagagaacgt
     2341 ataaggagtt tactccctat cagtgataga gaacgtatga ccagtttact ccctatcagt
     2401 gatagagaac gtatctacag tttactccct atcagtgata gagaacgtat atccagttta
     2461 ctccctatca gtgatagaga acgtataagc tttaggcgtg tacggtgggc gcctataaaa
     2521 gcagagctcg tttagtgaac cgtcagatcg cctggagcaa ttccacaaca cttttgtctt
     2581 ataccaactt tccgtaccac ttcctaccct cgtaaagaag ttcctattcc gaagttccta
     2641 ttcttcaaaa ggtataggaa cttcgctcga atgtcgatga gtccaaagca cacgactccg
     2701 ttctcagtgt ctgacatctt gagtcccctg gaggaaagct acaagaaagt gggcatggag
     2761 ggcggcggcc tcggggctcc gctggcggcg tacaggcagg gccaggcggc accgccaaca
     2821 gcggccatgc agcagcacgc cgtggggcac cacggcgccg tcaccgccgc ctaccacatg
     2881 acggcggcgg gggtgcccca gctctcgcac tccgccgtgg ggggctactg caacggcaac
     2941 ctgggcaaca tgagcgagct gccgccgtac caggacacca tgaggaacag cgcctctggc
     3001 cccggatggt acggcgccaa cccagacccg cgcttccccg ccatctcccg cttcatgggc
     3061 ccggcgagcg gcatgaacat gagcggcatg ggcggcctgg gctcgctggg ggacgtgagc
     3121 aagaacatgg ccccgctgcc aagcgcgccg cgcaggaagc gccgggtgct cttctcgcag
     3181 gcgcaggtgt acgagctgga gcgacgcttc aagcaacaga agtacctgtc ggcgccggag
     3241 cgcgagcacc tggccagcat gatccacctg acgcccacgc aggtcaagat ctggttccag
     3301 aaccaccgct acaaaatgaa gcgccaggcc aaggacaagg cggcgcagca gcaactgcag
     3361 caggacagcg gcggcggcgg gggcggcggg ggcaccgggt gcccgcagca gcaacaggct
     3421 cagcagcagt cgccgcgacg cgtggcggtg ccggtcctgg tgaaagacgg caaaccgtgc
     3481 caggcgggtg cccccgcgcc gggcgccgcc agcctacaag gccacgcgca gcagcaggcg
     3541 cagcaccagg cgcaggccgc gcaggcggcg gcagcggcca tctccgtggg cagcggtggc
     3601 gccggccttg gcgcacaccc gggccaccag ccaggcagcg caggccagtc tccggacctg
     3661 gcgcaccacg ccgccagccc cgcggcgctg cagggccagg tatccagcct gtcccacctg
     3721 aactcctcgg gctcggacta cggcaccatg tcctgctcca ccttgctata cggtcggacc
     3781 tggtgaggcc gcctaatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     3841 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     3901 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     3961 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     4021 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttacata
     4081 tgactagtaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc
     4141 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg
     4201 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     4261 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     4321 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct
     4381 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     4441 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     4501 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     4561 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg
     4621 tgaccggcgc ctacacctgc agccaacgcc accatggtgt ctaagggcga agagctgatt
     4681 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaacca ccacttcaag
     4741 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg
     4801 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctaccagctt catgtacggc
     4861 agcagaacct tcatcaacca cacccagggc atccccgact tctttaagca gtccttccct
     4921 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc
     4981 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
     5041 ttcccatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc caacaccgag
     5101 atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg acatggccct gaagctcgtg
     5161 ggcgggggcc acctgatctg caacttcaag accacataca gatccaagaa acccgctaag
     5221 aacctcaaga tgcccggcgt ctactatgtg gaccacagac tggaaagaat caaggaggcc
     5281 gacaaagaga cctacgtcga gcagcacgag gtggctgtgg ccagatactg cgacctccct
     5341 agcaaactgg ggcacaaact taatgctagc gagggcagag gaagtctgct aacatgcggt
     5401 gacgtcgagg agaatcctgg cccaatgtct agactggaca agagcaaagt cataaactct
     5461 gctctggaat tactcaatgg agtcggtatc gaaggcctga cgacaaggaa actcgctcaa
     5521 aagctgggag ttgagcagcc taccctgtac tggcacgtga agaacaagcg ggccctgctc
     5581 gatgccctgc caatcgagat gctggacagg catcataccc actcctgccc cctggaaggc
     5641 gagtcatggc aagactttct gcggaacaac gccaagtcat accgctgtgc tctcctctca
     5701 catcgcgacg gggctaaagt gcatctcggc acccgcccaa cagagaaaca gtacgaaacc
     5761 ctggaaaatc agctcgcgtt cctgtgtcag caaggcttct ccctggagaa cgcactgtac
     5821 gctctgtccg ccgtgggcca ctttacactg ggctgcgtat tggaggaaca ggagcatcaa
     5881 gtagcaaaag aggaaagaga gacacctacc accgattcta tgcccccact tctgaaacaa
     5941 gcaattgagc tgttcgaccg gcagggagcc gaacctgcct tccttttcgg cctggaacta
     6001 atcatatgtg gcctggagaa acagctaaag tgcgaaagcg gcgggccgac cgacgccctt
     6061 gacgattttg acttagacat gctcccagcc gatgcccttg acgactttga ccttgatatg
     6121 ctgcctgctg acgctcttga cgattttgac cttgacatgc tccccgggta agtcgacgat
     6181 cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     6241 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     6301 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     6361 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     6421 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     6481 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     6541 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     6601 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     6661 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     6721 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctgagatc
     6781 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     6841 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     6901 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     6961 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7021 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7081 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     7141 agagtgagag gcccgggtta attaaggaaa gggctagatc attcttgaag acgaaagggc
     7201 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     7261 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     7321 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7381 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7441 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     7501 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7561 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7621 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7681 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7741 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7801 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     7861 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7921 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7981 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     8041 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     8101 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     8161 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     8221 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     8281 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     8341 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8401 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     8461 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     8521 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     8581 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     8641 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     8701 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8761 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     8821 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     8881 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8941 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     9001 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     9061 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     9121 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     9181 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     9241 tgcagcaagc tcatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     9301 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     9361 agctccccgt ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     9421 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     9481 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgacatgatt
     9541 acgaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     9601 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     9661 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     9721 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     9781 acaaacttag tagt
//
