LOCUS       sequence-234728-       10583 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Barcoded lentiviral vector to express KLF8 in mammalian cells under 
            control of EF1a promoter. Barcoded for transcript detection in 
            single cell RNA-seq..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10583)
  AUTHORS   Parekh U, Wu Y, Zhao D, Worlikar A, Shah N, Zhang K, Mali P
  TITLE     Mapping Cellular Reprogramming via Pooled Overexpression Screens 
            with Paired Fitness and Single-Cell RNA-Sequencing Readout.
  JOURNAL   Cell Syst. 2018 Nov 12. pii: S2405-4712(18)30433-2. doi: 
            10.1016/j.cels.2018.10.008.
  PUBMED    30448000
REFERENCE   2  (bases 1 to 10583)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Syst. 
            2018 Nov 12. pii: S2405-4712(18)30433-2. doi: 
            10.1016/j.cels.2018.10.008."
COMMENT     Sequence Label: EF1a_KLF8_P2A_Hygro_Barcode
FEATURES             Location/Qualifiers
     source          1..10583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2693..3871
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2924..3862
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3819..3839
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     3888..3907
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3888..3906
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3964..3973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5056..5112
                     /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             5113..6138
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5113..6138
                     /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"
     protein_bind    complement(6330..6363)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             6388..6568
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6594..6611)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6600..6824
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6870..7298
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6957..6976)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7167..7188
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7307..7327)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7312..7641
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7492..7627
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7554..7573
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7689..7736
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7755..8129
                     /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    8259..8380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8296..8315)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8350..8369
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8429..8445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8429..8445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8442..8464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8453..8469
                     /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(8477..8483,8484..8501,8502..8507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8522..8543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8660..8677)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8831..9419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8911..8930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9590..10381,10382..10450))
                     /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     10213..10232
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10451..10555)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta atcgataagc tttgcaaaga tggataaagt tttaaacaga
     2641 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
     2701 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     2761 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     2821 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     2881 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
     2941 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
     3001 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
     3061 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
     3121 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
     3181 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     3241 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     3301 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     3361 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     3421 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     3481 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     3541 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     3601 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     3661 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     3721 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     3781 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     3841 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac
     3901 tatagggaga cccaagctgg ctaggtaagc ttgatatcga attcctgcag cccgggggat
     3961 cccgccacca tggtcgatat ggataaactc ataaacaact tggaggtcca acttaattca
     4021 gaaggtggct caatgcaggt attcaagcag gtcactgctt ctgttcggaa cagagatccc
     4081 cctgagatag aatacagaag taatatgact tctccaacac tcctggatgc caaccccatg
     4141 gagaacccag cactgtttaa tgacatcaag attgagcccc cagaagaact tttggctagt
     4201 gatttcagcc tgccccaagt ggaaccagtt gacctctcct ttcacaagcc caaggctcct
     4261 ctccagcctg ctagcatgct acaagctcca atacgtcccc ccaagccaca gtcttctccc
     4321 cagacccttg tggtgtccac gtcaacatct gacatgagca cttcagcaaa cattcctact
     4381 gttctgaccc caggctctgt cctgacctcc tctcagagca ctggtagcca gcagatctta
     4441 catgtcattc acactatccc ctcagtcagt ctgccaaata agatgggtgg cctgaagacc
     4501 atcccagtgg tagtgcagtc tctgcccatg gtgtatacta ctttgcctgc agatgggggc
     4561 cctgcagcca ttacagtccc actcattgga ggagatggta aaaatgctgg atcagtgaaa
     4621 gttgacccca cctccatgtc tccactggaa attccaagtg acagtgagga gagtacaatt
     4681 gagagtggat cctcagcctt gcagagtctg cagggactac agcaagaacc agcagcaatg
     4741 gcccaaatgc agggagaaga gtcgcttgac ttgaagagaa gacggattca ccaatgtgac
     4801 tttgcaggat gcagcaaagt gtacaccaaa agctctcacc tgaaagctca ccgcagaatc
     4861 catacaggag agaagcctta taaatgcacc tgggatggct gctcctggaa atttgctcgc
     4921 tcagatgagc tcactcgcca tttccgcaag cacacaggca tcaagccttt tcggtgcaca
     4981 gactgcaacc gcagcttttc tcgttctgac cacctgtccc tgcatcgccg tcgccatgac
     5041 accatgggat ccggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag
     5101 aatcctggac cgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc
     5161 gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct
     5221 ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt
     5281 ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa
     5341 gtgcttgaca ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag
     5401 ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg
     5461 gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc
     5521 ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat
     5581 ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag
     5641 gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac
     5701 gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac
     5761 tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg
     5821 ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt
     5881 gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag
     5941 agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc
     6001 gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc
     6061 tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt
     6121 ccgagggcaa aggaataagt tagaactatt tcctggctgt tacgcgttaa cacagtaccg
     6181 aatgacctac tgacgggctt gttaaacagt gaaccgcccg ccccacgacc cgcagcgccc
     6241 gaccgaaagg agcgcacgac cccatgcatc gaacaaaaga aaagggggga ctggaagggc
     6301 taattcactc ccaacgaaga caagatatca taacttcgta tagcatacat tatacgaagt
     6361 tatcggctag ctggtccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg
     6421 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     6481 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     6541 ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac
     6601 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     6661 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     6721 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     6781 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     6841 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     6901 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     6961 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     7021 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     7081 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     7141 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     7201 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     7261 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     7321 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     7381 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     7441 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     7501 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     7561 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     7621 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     7681 cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     7741 tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg
     7801 tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg
     7861 acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc
     7921 aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg
     7981 ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg
     8041 agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg
     8101 tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg
     8161 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     8221 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     8281 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     8341 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     8401 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     8461 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     8521 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     8581 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     8641 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     8701 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     8761 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     8821 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     8881 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8941 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     9001 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     9061 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     9121 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     9181 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     9241 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     9301 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     9361 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     9421 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     9481 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     9541 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     9601 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     9661 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     9721 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     9781 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     9841 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     9901 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     9961 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
    10021 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
    10081 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
    10141 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
    10201 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
    10261 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
    10321 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
    10381 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
    10441 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
    10501 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
    10561 ttccccgaaa agtgccacct gac
//
