LOCUS       sequence-353438-       10789 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral vector for overexpressing transcription factor ORFs with 
            unique 24-bp barcodes. Barcodes facilitate identification of 
            transcription factors in pooled screens..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10789)
  AUTHORS   Joung J, Ma S, Tay T, Geiger-Schuller KR, Kirchgatterer PC, Verdine 
            VK, Guo B, Arias-Garcia MA, Allen WE, Singh A, Kuksenko O, Abudayyeh
            OO, Gootenberg JS, Fu Z, Macrae RK, Buenrostro JD, Regev A, Zhang F
  TITLE     A transcription factor atlas of directed differentiation.
  JOURNAL   Cell. 2023 Jan 5;186(1):209-229.e26. doi: 
            10.1016/j.cell.2022.11.026.
  PUBMED    36608654
REFERENCE   2  (bases 1 to 10789)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2022.11"; journalName: "Cell"; date: "2023-01-5- 5";
            volume: "186"; issue: "1"; pages: "209-229.e26"
COMMENT     Sequence Label: TFORF0473
FEATURES             Location/Qualifiers
     source          1..10789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1190..1199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3727..4315
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3780..3800)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4198..4209)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4387..4620
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4698..4819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4735..4754)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4789..4808
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4859..4994
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5014..5033)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5015..5033)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5043..5060)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5043..5059)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5052..5074)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5201..5656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5288..5307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5498..5519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5682..5786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5787..5855,5856..6647)
                     /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     complement(6005..6024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6818..7406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7307..7326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7560..7577
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7694..7715
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7730..7735,7736..7753,7754..7760)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7768..7784
                     /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)."
     primer_bind     7773..7795
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7792..7808
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7792..7808
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7827..7847
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7829..7847
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7875..8101
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             8102..8282
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8329..8454
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8947..9180
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9365..9409
                     /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"
     promoter        9640..9850
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        9662..9858
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9709..9844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             9890..10492
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGAS"
     primer_bind     complement(9890..9909)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10386..10406
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10535..10652
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gctagcaccg ccaccatggc
     1201 ccagcagggc cagcaggggc agatggcaag tggagacagc aatctcaact tcaggatggt
     1261 cgcagagatc cagaacgtag agggtcagaa cctgcaggag caggtttgcc ctgagcccat
     1321 tttcaggttc ttcagagaaa acaaggtgga gattgcaagt gcaataacaa ggccatttcc
     1381 tttccttatg ggcctccgag accgctcctt catctccgag cagatgtatg aacattttca
     1441 agaagctttt agaaacctgg tcccagtgac aagagtgatg tattgtgtac tcagtgaact
     1501 ggagaagaca tttggctggt cacatctgga agcattgttc agcaggatta acctgatggc
     1561 ctatcctgat ttaaacgaga tttacagaag cttccagaat gtatgctatg aacactcacc
     1621 tctccaaatg aataatgtaa acgatttaga agatagaccc agattactac catatggtaa
     1681 acaagagaac agcaatgcct gtcatgaaat ggatgatata gcagtgcctc aggaagcctt
     1741 gagctcctcg ccaaggtgtg agccaggttt ctcttcagag tcttgtgagc agttagctct
     1801 cccaaaggct ggtggaggag atgctgaaga tgcacccagc ctactaccag gtgggggagt
     1861 tctagaaagc aacgggatga tagatgcggc aaggacatac agcacagcac caggggagaa
     1921 acagggagag gaggaaggca ggaacagtcc cagaaaaaga aaccaagaca aggagaagta
     1981 ccaagagagt ccagagggaa gagacaaaga gacctttgat ctaaaaactc cccaagtcac
     2041 taatgaagga gaaccagaga aggggctctg tctactacca ggtgaaggag aagagggcag
     2101 tgatgactgt tcagaaatgt gtgatggaga agagccccag gaagcctcta gctccctagc
     2161 aagatgtggg tcagagggca gtgatgactg ttcggaaatg tgtgatggag aagagcgcca
     2221 ggaagcctct agctccctag caagacgtgg gtcagtgtct agtgaactag aaaatcaccc
     2281 aatgaatgaa gaaggagaat cagaagagct tgcttctagc ctgctatatg ataatgtacc
     2341 aggagcggag caatcagcat atgaaaatga gaagtgttcc tgtgtcatgt gtttctcaga
     2401 agaggtgcca ggaagcccag aagcaaggac ggaaagtgat caagcgtgtg gcacaatgga
     2461 tactgtggat attgcaaaca actccacttt gggaaaaccc aagaggaaaa gaagaaaaaa
     2521 gagggggcat ggctggagca gaatgagaat gagaaggcag gaaaacagcc aacaaaatga
     2581 taatagcaaa gccgacggcc aggtggtctc cagtgaaaag aaggcgaacg tgaatctgaa
     2641 agacctttcc aagattaggg ggagaaagag aggcaaacct ggaacccgct tcactcagag
     2701 tgacagagct gcacagaaaa gagtccgatc aagagcttca agaaagcaca aagatgaaac
     2761 tgtggatttt aaggctcctt tgcttccagt gacctgtggt ggggtgaagg gaattttaca
     2821 taagaagaaa ttgcagcaag gaatcttggt gaagtgtata cagactgagg atggaaaatg
     2881 gttcaccccc acggaatttg aaatcaaagg aggccatgca agatcaaaga actggaggct
     2941 gagtgtgcgc tgtggcgggt ggcccctacg atggctgatg gagaatggat ttctgcctga
     3001 tcctccaaga atacgttaca ggaaaaaaaa gagaatactg aagtctcaaa acaatagctc
     3061 agttgaccct tgtatgagaa acctggatga gtgtgaggtg tgccgggacg gaggggagct
     3121 gttctgttgc gacacttgtt caagagtctt ccatgaggac tgtcacatcc cgcctgtgga
     3181 agctgagagg accccgtgga attgcatctt ctgcaggatg aaggagtctc cgggaagcca
     3241 acagtgttgt caggaatctg aggtcctgga gaggcagatg tgtcctgagg aacagttgaa
     3301 atgtgagttc ctcctcttga aagtctattg ctgttctgag agctcctttt ttgccaagat
     3361 tccatactat tattatatta gagaggcgtg tcaaggcctg aaggagccca tgtggttgga
     3421 taaaatcaag aaaaggctga atgagcacgg ttacccccaa gtggaggggt ttgtacaaga
     3481 catgcgcctc atcttccaga accacagggc ctcttacaag tacaaggatt ttggccaaat
     3541 gggatttaga ctggaggctg agtttgagaa gaatttcaag gaagtgtttg ctattcagga
     3601 aacaaatggg aacaattagt aatgaactag tccgtaactt gaaagtattt cgatttcttg
     3661 gctttatata tcttgtggaa aggacgatta cagtctgcta ccggtttaaa tacgcgttaa
     3721 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3781 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3841 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3901 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3961 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4021 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4081 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     4141 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4201 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4261 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga
     4321 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     4381 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     4441 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     4501 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     4561 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     4621 gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     4681 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4741 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4801 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     4861 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4921 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4981 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     5041 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     5101 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5161 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     5221 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5281 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5341 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5401 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5461 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5521 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     5581 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5641 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5701 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5761 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5881 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5941 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6001 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6061 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6181 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6301 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6361 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6481 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6661 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6721 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6781 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6841 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6901 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6961 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7021 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7081 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7141 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7201 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7261 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7321 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7381 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7441 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7501 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     7561 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     7621 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7681 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     7741 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     7801 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     7861 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     7921 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     7981 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     8041 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     8101 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     8161 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     8221 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     8281 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     8341 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     8401 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     8461 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     8521 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     8581 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     8641 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     8701 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     8761 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     8821 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     8881 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     8941 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     9001 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     9061 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     9121 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     9181 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     9241 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     9301 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     9361 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     9421 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     9481 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     9541 accattatcg tttcagaccc acctcccaac cccgagggga cccatgcatt gcatctcaat
     9601 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     9661 atgcgtctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     9721 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     9781 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     9841 ggcctaggct tttgcaaaaa gctttctaga ggtaccaccg gagcttacca tgaccgagta
     9901 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc
     9961 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga
    10021 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
    10081 gtgggtcgcg gacgacggcg ccgccgtggc ggtctggacc acgccggaga gcgtcgaagc
    10141 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
    10201 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
    10261 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
    10321 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
    10381 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
    10441 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgccagtt aacgaattct
    10501 agatcttgag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
    10561 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
    10621 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
    10681 cactttggcg ccggctcgag gcctgcaggt gcaaagatgg ataaagtttt aaacagagag
    10741 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaatt
//
