LOCUS       sequence-234532-       11402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Barcoded lentiviral vector to express HNF1A 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 11402)
  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 11402)
  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_HNF1A_P2A_Hygro_Barcode
FEATURES             Location/Qualifiers
     source          1..11402
                     /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"
     regulatory      5439..5448
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5874..5930
                     /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             5931..6956
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             5931..6956
                     /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(7148..7181)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             7206..7386
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7412..7429)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7418..7642
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7688..8116
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7775..7794)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7985..8006
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8125..8145)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8130..8459
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8310..8445
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8372..8391
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8507..8554
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8573..8947
                     /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    9077..9198
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9114..9133)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9168..9187
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9247..9263)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9247..9263)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9260..9282)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9271..9287
                     /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(9295..9301,9302..9319,9320..9325))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9340..9361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9478..9495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9649..10237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9729..9748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10408..11199,11200..11268))
                     /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     11031..11050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11269..11373)
                     /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 ggtttctaaa ctgagccagc tgcagacgga gctcctggcg gccctgctgg
     4021 agtcagggct gagcaaagag gcactgctcc aggcactggg tgagccgggg ccctacctcc
     4081 tggctggaga aggccccctg gacaaggggg agtcctgcgg cggcggtcga ggggagctgg
     4141 ctgagctgcc caatgggctg ggggagactc ggggctccga ggacgagacg gacgacgatg
     4201 gggaagactt cacgccaccc atcctcaaag agctggagaa cctcagccct gaggaggcgg
     4261 cccaccagaa agccgtggtg gagacccttc tgcaggagga cccgtggcgt gtggcgaaga
     4321 tggtcaagtc ctacctgcag cagcacaaca tcccacagcg ggaggtggtc gataccactg
     4381 gcctcaacca gtcccacctg tcccaacacc tcaacaaggg cactcccatg aagacgcaga
     4441 agcgggccgc cctgtacacc tggtacgtcc gcaagcagcg agaggtggcg cagcagttca
     4501 cccatgcagg gcagggaggg ctgattgaag agcccacagg tgatgagcta ccaaccaaga
     4561 aggggcggag gaaccgtttc aagtggggcc cagcatccca gcagatcctg ttccaggcct
     4621 atgagaggca gaagaaccct agcaaggagg agcgagagac gctagtggag gagtgcaata
     4681 gggcggaatg catccagaga ggggtgtccc catcacaggc acaggggctg ggctccaacc
     4741 tcgtcacgga ggtgcgtgtc tacaactggt ttgccaaccg gcgcaaagaa gaagccttcc
     4801 ggcacaagct ggccatggac acgtacagcg ggcccccccc agggccaggc ccgggacctg
     4861 cgctgcccgc tcacagctcc cctggcctgc ctccacctgc cctctccccc agtaaggtcc
     4921 acggtgtgcg ctatggacag cctgcgacca gtgagactgc agaagtaccc tcaagcagcg
     4981 gcggtccctt agtgacagtg tctacacccc tccaccaagt gtcccccacg ggcctggagc
     5041 ccagccacag cctgctgagt acagaagcca agctggtctc agcagctggg ggccccctcc
     5101 cccctgtcag caccctgaca gcactgcaca gcttggagca gacatcccca ggcctcaacc
     5161 agcagcccca gaacctcatc atggcctcac ttcctggggt catgaccatc gggcctggtg
     5221 agcctgcctc cctgggtcct acgttcacca acacaggtgc ctccaccctg gtcatcggcc
     5281 tggcctccac gcaggcacag agtgtgccgg tcatcaacag catgggcagc agcctgacca
     5341 ccctgcagcc cgtccagttc tcccagccgc tgcacccctc ctaccagcag ccgctcatgc
     5401 cacctgtgca gagccatgtg acccagagcc ccttcatggc caccatggct cagctgcaga
     5461 gcccccacgc cctctacagc cacaagcccg aggtggccca gtacacccac acaggcctgc
     5521 tcccgcagac tatgctcatc accgacacca ccaacctgag cgccctggcc agcctcacgc
     5581 ccaccaagca ggtcttcacc tcagacactg aggcctccag tgagtccggg cttcacacgc
     5641 cggcatctca ggccaccacc ctccacgtcc ccagccagga ccctgccggc atccagcacc
     5701 tgcagccggc ccaccggctc agcgccagcc ccacagtgtc ctccagcagc ctggtgctgt
     5761 accagagctc agactccagc aatggccaga gccacctgct gccatccaac cacagcgtca
     5821 tcgagacctt catctccacc cagatggcct cttcctccca gttgggatcc ggcgcaacaa
     5881 acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg atgaaaaagc
     5941 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     6001 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     6061 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     6121 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     6181 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     6241 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     6301 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     6361 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     6421 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     6481 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     6541 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     6601 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     6661 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     6721 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     6781 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     6841 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     6901 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataagtta
     6961 gaactatttc ctggctgtta cgcggcgttt ggcgctaatg tgatctactg acgggcttgt
     7021 taaacagtga accgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     7081 catgcatcga acaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     7141 agatatcata acttcgtata gcatacatta tacgaagtta tcggctagct ggtccggact
     7201 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     7261 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     7321 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     7381 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     7441 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     7501 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     7561 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     7621 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     7681 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     7741 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     7801 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     7861 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     7921 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     7981 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     8041 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     8101 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     8161 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     8221 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     8281 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     8341 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     8401 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     8461 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc
     8521 atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa
     8581 gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg
     8641 gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg
     8701 ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct
     8761 ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc
     8821 cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg
     8881 gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca
     8941 ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     9001 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     9061 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     9121 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     9181 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     9241 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     9301 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     9361 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     9421 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     9481 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     9541 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     9601 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     9661 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     9721 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     9781 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     9841 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     9901 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     9961 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
    10021 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
    10081 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
    10141 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
    10201 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
    10261 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
    10321 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
    10381 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
    10441 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
    10501 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
    10561 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
    10621 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
    10681 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
    10741 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
    10801 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
    10861 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
    10921 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
    10981 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
    11041 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
    11101 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
    11161 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
    11221 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
    11281 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
    11341 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
    11401 cg
//
