LOCUS       sequence-234723-       10541 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Barcoded lentiviral vector to express KLF3 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 10541)
  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 10541)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_KLF3_P2A_Hygro_Barcode
FEATURES             Location/Qualifiers
     source          1..10541
                     /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"
     CDS             5014..5070
                     /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             5071..6096
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5071..6096
                     /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(6288..6321)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             6346..6526
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6552..6569)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6558..6782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6828..7256
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6915..6934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7125..7146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7265..7285)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7270..7599
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7450..7585
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7512..7531
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7647..7694
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7713..8087
                     /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    8217..8338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8254..8273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8308..8327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8387..8403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8387..8403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8400..8422)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8411..8427
                     /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(8435..8441,8442..8459,8460..8465))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8480..8501
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8618..8635)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8789..9377)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8869..8888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9548..10339,10340..10408))
                     /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     10171..10190
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10409..10513)
                     /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 tgctcatgtt tgacccagtt cctgtcaagc aagaggccat ggaccctgtc
     4021 tcagtgtcat acccatctaa ttacatggaa tccatgaagc ctaacaagta tggggtcatc
     4081 tactccacac cattgcctga gaagttcttt cagaccccag aaggtctgtc gcacggaata
     4141 cagatggagc cagtggacct cacggtgaac aagcggagtt cacccccttc ggctgggaat
     4201 tcgccctcct ctctgaagtt cccgtcctca caccggagag cctcgcctgg gttgagcatg
     4261 ccttcttcca gcccaccgat aaaaaaatac tcaccccctt ctccaggcgt gcagcccttc
     4321 ggcgtgccgc tgtccatgcc accagtgatg gcagctgccc tctcgcggca tggaatacgg
     4381 agcccgggga tcctgcccgt catccagccg gtggtggtgc agcccgtccc ctttatgtac
     4441 acaagtcacc tccagcagcc tctcatggtc tccttatcgg aggagatgga aaattccagt
     4501 agtagcatgc aagtacctgt aattgaatca tatgagaagc ctatatcaca gaaaaaaatt
     4561 aaaatagaac ctgggatcga accacagagg acagattatt atcctgaaga aatgtcaccc
     4621 cccttaatga actcagtgtc ccccccgcaa gcattgttgc aagagaatca cccttcggtc
     4681 atcgtgcagc ctgggaagag acctttacct gtggaatccc cggatactca aaggaagcgg
     4741 aggatacaca gatgtgatta tgatggatgc aacaaagtgt acactaaaag ctcccacttg
     4801 aaagcacaca gaagaacaca cacaggagaa aaaccctaca aatgtacatg ggaagggtgc
     4861 acatggaagt ttgctcggtc tgatgaacta acaagacatt tccgaaaaca tactggaatc
     4921 aaacctttcc agtgcccgga ctgtgaccgc agcttctccc gttctgacca tcttgccctc
     4981 cataggaaac gccacatgct agtcggatcc ggcgcaacaa acttctctct gctgaaacaa
     5041 gccggagatg tcgaagagaa tcctggaccg atgaaaaagc ctgaactcac cgcgacgtct
     5101 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     5161 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     5221 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     5281 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
     5341 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     5401 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
     5461 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     5521 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     5581 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     5641 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     5701 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     5761 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     5821 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     5881 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     5941 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     6001 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     6061 cgacgcccca gcactcgtcc gagggcaaag gaataagtta gaactatttc ctggctgtta
     6121 cgcgtcgcag agtccacaca agtcctactg acgggcttgt taaacagtga accgcccgcc
     6181 ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catgcatcga acaaaagaaa
     6241 aggggggact ggaagggcta attcactccc aacgaagaca agatatcata acttcgtata
     6301 gcatacatta tacgaagtta tcggctagct ggtccggact gtactgggtc tctctggtta
     6361 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     6421 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     6481 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac
     6541 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     6601 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     6661 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     6721 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     6781 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     6841 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     6901 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     6961 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     7021 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     7081 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7141 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     7201 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     7261 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     7321 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     7381 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     7441 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     7501 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     7561 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     7621 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     7681 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     7741 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     7801 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     7861 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     7921 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     7981 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     8041 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     8101 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     8161 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     8221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     8281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     8341 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     8401 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     8461 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     8521 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     8581 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     8641 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     8701 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     8761 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     8821 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     8881 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     8941 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     9001 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     9061 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     9121 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     9181 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     9241 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     9301 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     9361 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     9421 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     9481 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     9541 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     9601 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     9661 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     9721 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     9781 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     9841 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     9901 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     9961 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
    10021 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
    10081 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
    10141 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
    10201 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
    10261 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
    10321 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
    10381 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
    10441 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
    10501 taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
