LOCUS       sequence-234480-       10214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded lentiviral vector to express ASCL1 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 10214)
  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 10214)
  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: "Cell Syst. 
            2018 Nov 12. pii: S2405-4712(18)30433-2. doi: 
            10.1016/j.cels.2018.10.008."
COMMENT     Sequence Label: EF1a_ASCL1_P2A_Hygro_Barcode
FEATURES             Location/Qualifiers
     source          1..10214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1679..1912
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2097..2141
                     /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    2439..2556
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2692..3870
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2923..3861
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3818..3838
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     3887..3906
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3887..3905
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3963..3972
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4686..4742
                     /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             4743..5768
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4743..5768
                     /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(5960..5993)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             6018..6198
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6224..6241)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6230..6454
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6500..6928
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6587..6606)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6797..6818
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6937..6957)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6942..7271
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7122..7257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7184..7203
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7319..7366
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7385..7759
                     /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    7889..8010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7926..7945)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7980..7999
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8059..8075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8059..8075)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8072..8094)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8083..8099
                     /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(8107..8113,8114..8131,8132..8137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8152..8173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8290..8307)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8461..9049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8541..8560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9220..10011,10012..10080))
                     /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     9843..9862
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10081..10185)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
     1561 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     1621 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     1681 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     1741 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     1801 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     1861 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     1921 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     1981 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     2041 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     2101 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     2161 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     2221 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     2281 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     2341 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg
     2401 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     2461 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2521 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2581 gatccagttt ggttaattaa tcgataagct ttgcaaagat ggataaagtt ttaaacagag
     2641 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt
     2701 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     2761 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2821 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2881 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
     2941 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
     3001 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
     3061 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
     3121 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
     3181 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
     3241 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
     3301 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
     3361 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
     3421 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     3481 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     3541 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     3601 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     3661 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     3721 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     3781 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     3841 agtttttttc ttccatttca ggtgtcgtga ggaattagct tggtactaat acgactcact
     3901 atagggagac ccaagctggc taggtaagct tgatatcgaa ttcctgcagc ccgggggatc
     3961 ccgccaccat ggagtcttct gctaaaatgg agtccggagg cgcgggacaa caaccacaac
     4021 cgcaaccaca acaacccttc ctgccgccgg ccgcatgttt tttcgcgacc gctgctgctg
     4081 ctgcagcggc ggcggctgct gccgccgcgc aatccgccca acagcaacaa caacaacagc
     4141 agcagcagca acaagcgcct caacttcgac ccgctgcaga cgggcagccc tcagggggag
     4201 ggcacaagag cgctccgaag caggttaaaa ggcagaggag cagtagtccc gaactgatgc
     4261 gatgtaagag gcgcctcaat tttagcggtt ttggttactc tttgccccag cagcagccgg
     4321 ctgccgtagc tcgccgaaat gagcgggaaa ggaaccgcgt taaacttgtg aatctcggtt
     4381 tcgcgacact tcgagagcac gtaccaaatg gggcagctaa caagaaaatg agtaaagttg
     4441 agacactgcg gtctgcagtg gagtatatta gagctcttca acaattgctt gacgagcacg
     4501 atgccgtatc agccgcattt caagccgggg tgctgtcccc aacaatatct ccgaactaca
     4561 gcaatgatct taatagcatg gcgggaagtc ccgtttcctc ctactcctct gatgagggca
     4621 gctacgaccc tctcagtccc gaggagcaag agcttcttga cttcactaac tggttcggat
     4681 ccggcgcaac aaacttctct ctgctgaaac aagccggaga tgtcgaagag aatcctggac
     4741 cgatgaaaaa gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg
     4801 acagcgtctc cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg
     4861 atgtaggagg gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag
     4921 atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca
     4981 ttggggaatt cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt
     5041 tgcaagacct gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg
     5101 atgcgatcgc tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag
     5161 gaatcggtca atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt
     5221 atcactggca aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg
     5281 agctgatgct ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg
     5341 gctccaacaa tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg
     5401 cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg
     5461 cttgtatgga gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc
     5521 cgcggctccg ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg
     5581 acggcaattt cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg
     5641 gagccgggac tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg
     5701 gctgtgtaga agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa
     5761 aggaataagt tagaactatt tcctggctgt tacgcgttcg tgataaaact atctgcctac
     5821 tgacgggctt gttaaacagt gaaccgcccg ccccacgacc cgcagcgccc gaccgaaagg
     5881 agcgcacgac cccatgcatc gaacaaaaga aaagggggga ctggaagggc taattcactc
     5941 ccaacgaaga caagatatca taacttcgta tagcatacat tatacgaagt tatcggctag
     6001 ctggtccgga ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6061 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6121 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6181 tgtggaaaat ctctagcagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     6241 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     6301 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     6361 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     6421 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     6481 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     6541 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     6601 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     6661 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     6721 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     6781 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     6841 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     6901 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     6961 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     7021 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     7081 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     7141 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     7201 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     7261 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
     7321 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
     7381 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
     7441 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
     7501 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
     7561 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
     7621 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
     7681 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
     7741 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
     7801 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     7861 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     7921 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     7981 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     8041 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     8101 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     8161 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     8221 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     8281 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     8341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     8401 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     8461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     8521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     8581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     8641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     8701 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     8761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     8821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8881 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     8941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     9001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     9061 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     9121 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     9181 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     9241 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     9301 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     9361 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     9421 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     9481 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     9541 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     9601 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     9661 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     9721 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     9781 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     9841 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     9901 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     9961 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
    10021 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
    10081 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
    10141 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
    10201 agtgccacct gacg
//
