LOCUS       sequence-234498-       10022 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded lentiviral vector to express ASCL4 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 10022)
  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 10022)
  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_ASCL4_P2A_Hygro_Barcode
FEATURES             Location/Qualifiers
     source          1..10022
                     /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             4494..4550
                     /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             4551..5576
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4551..5576
                     /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(5768..5801)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             5826..6006
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6032..6049)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6038..6262
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6308..6736
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6395..6414)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6605..6626
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6745..6765)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6750..7079
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6930..7065
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6992..7011
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7127..7174
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7193..7567
                     /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    7697..7818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7734..7753)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7788..7807
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7867..7883)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7867..7883)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7880..7902)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7891..7907
                     /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(7915..7921,7922..7939,7940..7945))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7960..7981
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8098..8115)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8269..8857)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8349..8368)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9028..9819,9820..9888))
                     /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     9651..9670
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9889..9993)
                     /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 ggagacgcgt aaaccggcgg aacggctggc cttgccatac tcgctgcgca
     4021 ccgcgcccct gggcgttccg gggaccctgc ccggactccc gcggagggac cccctcaggg
     4081 tcgccctgcg tctggacgcc gcgtgctggg agtgggcgcg cagcggctgc gcacggggat
     4141 ggcagtactt gcccgtgccg ctggacagcg ccttcgagcc cgccttcctc cgcaagcgca
     4201 acgagcgcga gcggcagcgg gtgcgctgcg tgaacgaggg ctatgcgcgc ctccgagacc
     4261 acctgccccg ggagctggca gacaagcgcc tcagcaaagt ggagacgctc cgcgctgcca
     4321 tcgactacat caagcacctg caggagctgc tggagcgcca ggcctggggg ctcgagggcg
     4381 cggccggcgc cgtcccccag cgcagggcgg aatgcaacag cgacggggag tccaaggcct
     4441 cttcggcgcc ttcgcccagc agcgagcccg aggagggggg cagcggatcc ggcgcaacaa
     4501 acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg atgaaaaagc
     4561 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     4621 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     4681 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     4741 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     4801 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     4861 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     4921 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     4981 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     5041 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     5101 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     5161 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     5221 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     5281 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     5341 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     5401 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     5461 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     5521 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataagtta
     5581 gaactatttc ctggctgtta cgcggcgaat tattacaggc ctctctactg acgggcttgt
     5641 taaacagtga accgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     5701 catgcatcga acaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     5761 agatatcata acttcgtata gcatacatta tacgaagtta tcggctagct ggtccggact
     5821 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5881 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5941 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6001 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     6061 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     6121 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     6181 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     6241 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     6301 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     6361 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     6421 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     6481 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     6541 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     6601 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6661 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6721 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     6781 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     6841 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6901 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6961 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     7021 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     7081 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     7141 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     7201 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     7261 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     7321 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     7381 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     7441 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     7501 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     7561 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     7621 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     7681 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7741 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7801 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     7861 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     7921 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     7981 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     8041 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     8101 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     8161 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     8221 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     8281 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8341 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8401 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8461 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     8521 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8581 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8641 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8701 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8761 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8821 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8881 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     8941 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     9001 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     9061 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     9121 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     9181 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     9241 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     9301 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     9361 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     9421 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     9481 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     9541 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     9601 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     9661 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     9721 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     9781 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     9841 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     9901 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     9961 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    10021 cg
//
