LOCUS       sequence-221423-       11276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Nfib and hygromycin resistance gene under control of tetO 
            sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11276)
  AUTHORS   Canals I, Ginisty A, Quist E, Timmerman R, Fritze J, Miskinyte G, 
            Monni E, Hansen MG, Hidalgo I, Bryder D, Bengzon J, Ahlenius H
  TITLE     Rapid and efficient induction of functional astrocytes from human 
            pluripotent stem cells.
  JOURNAL   Nat Methods. 2018 Sep;15(9):693-696. doi: 10.1038/s41592-018-0103-2.
            Epub 2018 Aug 20.
  PUBMED    30127505
REFERENCE   2  (bases 1 to 11276)
  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"; doi: 
            "10.1038/s41592-018-0103-2"; journalName: "Nat Methods"; date: 
            "2018-09"; volume: "15"; issue: "9"; pages: "693-696"
COMMENT     Sequence Label: tetO.Nfib.Hygro
FEATURES             Location/Qualifiers
     source          1..11276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             604..784
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    831..956
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1449..1682
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1867..1911
                     /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    2209..2326
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    join(2383..2401,2402..2424,2425..2443,2444..2466,2467..2485,
                     2486..2508,2509..2527,2528..2550,2551..2569,2570..2592,
                     2593..2611,2612..2634,2635..2653)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        2686..2723
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2720..2744
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    4089..4656
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4106..4658
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4250..4267)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4477..4496
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4673..5698
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4673..5698
                     /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
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     misc_feature    5737..6325
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5790..5810)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6208..6219)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6328..6344)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6329..6345)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6850..7030
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7056..7073)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7062..7286
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7332..7760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7419..7438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7629..7650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7769..7789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7774..8103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7954..8089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8016..8035
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8151..8198
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8217..8591
                     /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"
     polyA_signal    8721..8842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8758..8777)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8812..8831
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8891..8907)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8891..8907)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8904..8926)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8915..8931
                     /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(8939..8945,8946..8963,8964..8969))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8984..9005
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9122..9139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9293..9881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9373..9392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10052..10843,10844..10912))
                     /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     10675..10694
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10913..11017)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11092..11111)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
      601 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
      661 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
      721 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
      781 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
      841 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
      901 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
      961 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
     1021 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
     1081 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
     1141 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
     1201 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
     1261 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
     1321 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     1381 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     1441 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     1501 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     1561 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     1621 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     1681 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     1741 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     1801 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     1861 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     1921 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     1981 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     2041 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     2101 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg
     2161 gcactgcgtg cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa
     2221 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     2281 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     2341 ggacagcaga gatccagttt ggttagatct cgagtttacc actccctatc agtgatagag
     2401 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
     2461 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
     2521 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     2581 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     2641 tcagtgatag agaaaagtga aagtcgagct cggtacccgg gtcgagtagg cgtgtacggt
     2701 gggaggccta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     2761 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc ggccccgaat
     2821 tcatgatgta ttctcccatc tgtctcactc aggatgaatt tcacccattt attgaggcac
     2881 ttcttcctca cgtccgtgca attgcctata cttggttcaa cctgcaggct cgaaaacgca
     2941 agtactttaa aaagcatgag aaacgaatgt cgaaggatga agaaagggca gtcaaagacg
     3001 agctgctcag tgagaagccc gaaatcaagc agaagtgggc atccaggctc ctggccaaac
     3061 tgcgcaaaga tatccgccag gagtaccggg aggactttgt gcttaccgtg actggcaaga
     3121 agcacccgtg ctgtgtctta tccaatccag accagaaggg taagattagg aggatcgact
     3181 gcctgcgaca ggcagacaaa gtctggcgtc tggatctagt catggtgatc ctgttcaaag
     3241 gcatcccttt ggagagtacg gatggagagc gactcatgaa gtccccgcac tgcacaaacc
     3301 cagcactttg tgttcagcca caccacatca cagtatcagt taaggagctt gacttgtttt
     3361 tggcatacta cgtgcaggag caagattctg gacaatcagg aagtccaagc cacagtgatc
     3421 ctgccaagaa tcctccaggg tacctcgagg acagctttgt aaaatccgga gtcttcaatg
     3481 tatcagagct tgtgagagta tccagaacac ccataaccca gggaactgga gtcaacttcc
     3541 caatcggaga aattcccagc caaccatact atcatgacat gaactctggt gtgaacctgc
     3601 agaggtcgct gtcttctcca ccgagcagca aaagacccaa aactatatct atagatgaaa
     3661 atatggagcc aagtcctaca ggagactttt acccctctcc aaattcacca gctgctggaa
     3721 gtcgaacatg gcatgaacga gatcaagata tgtcttctcc aactacaatg aagaagcctg
     3781 agaagccact gtttagctct acatctccac aggattcttc cccaagattg agcactttcc
     3841 cccagcacca tcatcccgga atacctggag tcgcgcacag tgtcatctca actcgaactc
     3901 cacctccgcc ctcaccgttg ccatttccga cgcaagctat ccttcctccg gcaccttcca
     3961 gctacttctc tcatccaaca atcagatatc ctcctcacct gaatcctcag gatactctga
     4021 agaactacgt accttcttat gacccatcca gtcctcaaac gagccagtcc tggtacctgg
     4081 gctagggatc cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg
     4141 tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc
     4201 cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa
     4261 ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga
     4321 caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc
     4381 ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc
     4441 cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac
     4501 aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg
     4561 tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac
     4621 ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacagcta gaatgaaaaa
     4681 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     4741 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     4801 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     4861 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     4921 tagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4981 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     5041 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     5101 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     5161 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     5221 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     5281 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     5341 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     5401 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     5461 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     5521 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     5581 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     5641 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagtt
     5701 aattaagcta gctaattcga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     5761 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5821 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5881 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5941 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     6001 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     6061 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     6121 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     6181 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     6241 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     6301 atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgagacct agaaaaacat
     6361 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     6421 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     6481 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     6541 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     6601 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     6661 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     6721 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     6781 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
     6841 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     6901 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     6961 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     7021 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     7081 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     7141 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     7201 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     7261 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     7321 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     7381 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     7441 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     7501 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     7561 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     7621 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     7681 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     7741 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     7801 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     7861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     7921 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     7981 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     8041 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     8101 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     8161 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     8221 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     8281 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     8341 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     8401 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     8461 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     8521 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     8581 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     8641 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     8701 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     8761 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     8821 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     8881 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     8941 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     9001 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     9061 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     9121 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     9181 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     9241 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     9301 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     9361 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     9421 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     9481 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     9541 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     9601 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     9661 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     9721 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     9781 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     9841 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     9901 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     9961 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    10021 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    10081 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    10141 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    10201 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    10261 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    10321 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    10381 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    10441 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    10501 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    10561 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    10621 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    10681 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    10741 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    10801 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    10861 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    10921 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    10981 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    11041 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
    11101 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
    11161 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
    11221 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatata
//
