LOCUS       sequence-334418-       11291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded lentiviral vector to express ERalpha (Y537S) 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 11291)
  AUTHORS   Parekh U, McDonald D, Dailamy A, Wu Y, Cordes T, Zhang K, Tipps A, 
            Metallo C, Mali P
  TITLE     Charting oncogenicity of genes and variants across lineages via 
            multiplexed screens in teratomas.
  JOURNAL   iScience. 2021 Sep 20;24(10):103149. doi: 
            10.1016/j.isci.2021.103149. eCollection 2021 Oct 22.
  PUBMED    34646987
REFERENCE   2  (bases 1 to 11291)
  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: "iScience.";
            date: "2021-09-20"; volume: "24(10):103149. doi"; pages: " 
            10.1016/j.isci.2021.103149. eCollection 2021 Oct 22"
COMMENT     Sequence Label: LV_EF1a_ERalpha(Y537S)-P2A-Hygro_Barcode
FEATURES             Location/Qualifiers
     source          1..11291
                     /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             4816..5751
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLSDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             4864..5754
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLSDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     CDS             5764..5820
                     /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             5821..6846
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5821..6846
                     /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(7038..7071)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             7096..7276
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7302..7319)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7308..7532
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7578..8006
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7665..7684)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7875..7896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8015..8035)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8020..8349
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8200..8335
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8262..8281
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8397..8444
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8463..8837
                     /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    8967..9088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9004..9023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9058..9077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9137..9153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9137..9153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9150..9172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9161..9177
                     /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(9185..9191,9192..9209,9210..9215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9230..9251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9368..9385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9539..10127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9619..9638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10298..11089,11090..11158))
                     /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     10921..10940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11159..11263)
                     /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 tgaccatgac cctccacacc aaagcatctg ggatggccct actgcatcag
     4021 atccaaggga acgagctgga gcccctgaac cgtccgcagc tcaagatccc cctggagcgg
     4081 cccctgggcg aggtgtacct ggacagcagc aagcccgccg tgtacaacta ccccgagggc
     4141 gccgcctacg agttcaacgc cgcggccgcc gccaacgcgc aggtctacgg tcagaccggc
     4201 ctcccctacg gccccgggtc tgaggctgcg gcgttcggct ccaacggcct ggggggtttc
     4261 cccccactca acagcgtgtc tccgagcccg ctgatgctac tgcacccgcc gccgcagctg
     4321 tcgcctttcc tgcagcccca cggccagcag gtgccctact acctggagaa cgagcccagc
     4381 ggctacacgg tgcgcgaggc cggcccgccg gcattctaca ggccaaattc agataatcga
     4441 cgccagggtg gcagagaaag attggccagt accaatgaca agggaagtat ggctatggaa
     4501 tctgccaagg agactcgcta ctgtgcagtg tgcaatgact atgcttcagg ctaccattat
     4561 ggagtctggt cctgtgaggg ctgcaaggcc ttcttcaaga gaagtattca aggacataac
     4621 gactatatgt gtccagccac caaccagtgc accattgata aaaacaggag gaagagctgc
     4681 caggcctgcc ggctccgcaa atgctacgaa gtgggaatga tgaaaggtgg gatacgaaaa
     4741 gaccgaagag gagggagaat gttgaaacac aagcgccaga gagatgatgg ggagggcagg
     4801 ggtgaagtgg ggtctgctgg agacatgaga gctgccaacc tttggccaag cccgctcatg
     4861 atcaaacgct ctaagaagaa cagcctggcc ttgtccctga cggccgacca gatggtcagt
     4921 gccttgttgg atgctgagcc ccccatactc tattccgagt atgatcctac cagacccttc
     4981 agtgaagctt cgatgatggg cttactgacc aacctggcag acagggagct ggttcacatg
     5041 atcaactggg cgaagagggt gccaggcttt gtggatttga ccctccatga tcaggtccac
     5101 cttctagaat gtgcctggct agagatcctg atgattggtc tcgtctggcg ctccatggag
     5161 cacccaggga agctactgtt tgctcctaac ttgctcttgg acaggaacca gggaaaatgt
     5221 gtagagggca tggtggagat cttcgacatg ctgctggcta catcatctcg gttccgcatg
     5281 atgaatctgc agggagagga gtttgtgtgc ctcaaatcta ttattttgct taattctgga
     5341 gtgtacacat ttctgtccag caccctgaag tctctggaag agaaggacca tatccaccga
     5401 gtcctggaca agatcacaga cactttgatc cacctgatgg ccaaggcagg cctgaccctg
     5461 cagcagcagc accagcggct ggcccagctc ctcctcatcc tctcccacat caggcacatg
     5521 agtaacaaag gcatggagca tctgtacagc atgaagtgca agaacgtggt gcccctctca
     5581 gatctgctgc tggagatgct ggacgcccac cgcctacatg cgcccactag ccgtggaggg
     5641 gcatccgtgg aggagacgga ccaaagccac ttggccactg cgggctctac ttcatcgcat
     5701 tccttgcaaa agtattacat cacgggggag gcagagggtt tccctgccac ggtcggatcc
     5761 ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg
     5821 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     5881 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     5941 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     6001 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     6061 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     6121 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     6181 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     6241 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     6301 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     6361 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     6421 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     6481 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     6541 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     6601 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     6661 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     6721 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     6781 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     6841 gaataagtta gaactatttc ctggctgtta cgcgtacccc acatggattg aatactactg
     6901 acgggcttgt taaacagtga accgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     6961 cgcacgaccc catgcatcga acaaaagaaa aggggggact ggaagggcta attcactccc
     7021 aacgaagaca agatatcata acttcgtata gcatacatta tacgaagtta tcggctagct
     7081 ggtccggact gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     7141 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     7201 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     7261 tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     7321 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     7381 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     7441 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     7501 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     7561 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     7621 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     7681 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     7741 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     7801 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     7861 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     7921 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     7981 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     8041 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     8101 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     8161 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     8221 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     8281 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     8341 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
     8401 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     8461 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
     8521 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
     8581 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
     8641 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
     8701 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
     8761 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
     8821 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     8881 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     8941 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     9001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     9061 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     9121 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     9181 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     9241 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     9301 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     9361 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     9421 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     9481 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     9541 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     9601 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     9661 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     9721 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     9781 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     9841 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     9901 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     9961 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    10021 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    10081 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    10141 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    10201 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    10261 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    10321 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    10381 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    10441 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    10501 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    10561 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    10621 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    10681 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    10741 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    10801 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    10861 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    10921 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    10981 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    11041 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    11101 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    11161 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    11221 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    11281 tgccacctga c
//
