LOCUS       sequence-475930-        8557 bp    DNA     circular SYN 01-JUL-2025
DEFINITION  Expresses V5-tagged CHIC2 (with K117R mutation) with Tomato 
            selectable marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8557)
  TITLE     A STUB1-CHIC2 complex inhibits CD8+ T cells to restrain tumor 
            immunity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8557)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pRRL-SFFV_V5-CHIC2-K117R_Tomato
FEATURES             Location/Qualifiers
     source          1..8557
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    296..421
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             468..648
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(649..875)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(903..923)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(903..921)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(942..958)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(942..958)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(955..977)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    966..982
                     /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(990..996,997..1014,1015..1020))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1035..1056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1173..1190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1344..1932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1424..1443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2103..2894,2895..2963))
                     /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     2726..2745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2964..3068)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3094..3549)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3231..3252)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3443..3462
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3676..3698
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3690..3707
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3691..3707
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3717..3736
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3717..3735
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3756..3891
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3931..4052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3942..3961)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3996..4015
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             4124..4357
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    4447..4850
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5085..5786)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(5168..5191)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(5366..5385)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5635..5653
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    join(5787..5795,5796..5798,5799..6373)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5960..5979)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     6189..6206
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    6431..6455
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(6952..6993)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(6996..7020)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(7074..7480)
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    7514..7631
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(7898..7942)
                     /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    8127..8360
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 tcttgtctaa agcttccttg gtgtctttta tctctatcct ttgatgcaca caatagaggg
       61 ttgctactgt attatataat gatctaagtt cttctgatcc tgtctgaagg gatggttgta
      121 gctgtcccag tatttgtcta cagccttctg atgtttctaa caggccagga ttaactgcga
      181 atcgttctag ctccctgctt gcccatacta tatgttttaa tttatatttt ttctttcccc
      241 ctggccttaa ccgaattttt tcccatcgcg atctaattct cccccgctta atactgacgc
      301 tctcgcaccc atctctctcc ttctagcctc cgctagtcaa aatttttggc gtactcacca
      361 gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag ccgagtcctg cgtcgagaga
      421 gctcctctgg tttccctttc gctttcaagt ccctgttcgg gcgccactgc tagagatttt
      481 ccacactgac taaaagggtc tgagggatct ctagttacca gagtcacaca acagacgggc
      541 acacactact tgaagcactc aaggcaagct ttattgaggc ttaagcagtg ggttccctag
      601 ttagccagag agctcccagg ctcagatctg gtctaaccag agagacccgt ttatgtatcg
      661 agctaggcac ttaaatacaa tatctctgca atgcggcaat tcagtggttc gtccaatcca
      721 tgtcagaccc gtctgttgcc ttcctaataa ggcacgatcg taccacctta cttccaccaa
      781 tcggcatgca cggtgctttt tctctccttg taaggcatgt tgctaactca tcgttaccat
      841 gttgcaagac tacaagagta ttgcataaga ctacattaag cttgcagctc cagcttttgt
      901 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
      961 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1021 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     1081 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     1141 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     1201 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1261 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1321 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1381 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1441 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1501 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1561 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1621 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1681 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1741 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1801 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1861 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1921 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1981 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2041 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2101 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2161 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2221 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2281 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2341 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2401 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2461 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2521 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2581 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2641 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2701 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2761 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2821 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2881 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2941 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3001 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3061 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     3121 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     3181 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     3241 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     3301 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     3361 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3421 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3481 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3541 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3601 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     3661 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat
     3721 acgactcact atagggcgaa ttgggtacgt ccctaggcct ccaaaaaagc ctcctcacta
     3781 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
     3841 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     3901 taggggcggg atagctagag ccagacatga taagatacat tgatgagttt ggacaaacca
     3961 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4021 ttgtaaccat tataagctgc aataaacaag ttcctctcac tctctgatat tcatttcttt
     4081 gcaagttata aatactgaat aataagatga catgaactac tactgctaga gattttccac
     4141 actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac
     4201 actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag
     4261 ccagagagct cccaggctca gatctggtct aaccagagag acccagtaca agcaaaaagc
     4321 agatcttgtc ttcgttggga gttacgtagc ccttccagtc cccccttttc ttttaaaaag
     4381 tggctaagat ctacagctgc cttgtaagtc attggtctta aaggtacctg atatcgaatt
     4441 cgaaaggacg tcagcttccc cgacaacacc acggaattat cagtgcccag caacctagcc
     4501 cctgtccagc agcgggcaag gcaggcggcg atgagttctg ccgtggcgat cgggaggggg
     4561 aaagcgaaag tcccagaaag gagttgacag gtggtggcaa tgccccagcc agtgggggtt
     4621 gcgtcagcaa acacagagca caccacgcca cgttgacgga caacgggcca caactcctca
     4681 taaagagaca gcaaccagga tttatacaag gaggagaaaa cgaaagccgt acgggaagca
     4741 atagcatgat acagaggcat taaagcagca tatccacaca gcgtaaaagg agcaacatag
     4801 ttaagaatat cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt aacaggaaca
     4861 gagcgtaaat aacgggaaag tttcttaaca tgtttgtctt gtggcaatac acctgaacta
     4921 gtaattacat atccctaaaa atgtaaatga ttgccccacc attttgtttt attaacattt
     4981 aaatgtatac ccaaatcaag aaaaacagaa caaatatggg aataaatggc ggtaagatgc
     5041 tcgaattcct gcagcccggg ggatccgtcg acgcggccgc tttacttgta cagctcgtcc
     5101 atgccgtaca ggaacaggtg gtggcggccc tcggagcgct cgtactgttc cacgatggtg
     5161 tagtcctcgt tgtgggaggt gatgtccagc ttggtgtcca cgtagtagta gccgggcagt
     5221 tgcacgggct tcttggccat gtagatggtc ttgaactcca ccaggtagtg gccgccgtcc
     5281 ttcagcttca gggcctggtg gatctcgccc ttcagcacgc cgtcgcgggg gtacaggcgc
     5341 tcggtggagg cctcccagcc catggtcttc ttctgcatta cggggccgtc gggggggaag
     5401 ttggtgccgc gcatcttcac cttgtagatc agcgtgccgt cctgcaggga ggagtcctgg
     5461 gtcacggtca ccagaccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg
     5521 gggaaggaca gcttcttgta atcggggatg tcggcggggt gcttcacgta cgccttggag
     5581 ccgtacatga actgggggga caggatgtcc caggcgaagg gcagggggcc gcccttggtc
     5641 accttcagct tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc
     5701 tcgaactcgt ggccgttcat ggagccctcc atgcgcacct tgaagcgcat gaactctttg
     5761 atgacctcct cgcccttgct caccatggtt gtggccatat tatcatcgtg tttttcaaag
     5821 gaaaaccacg tccccgtggt tcggggggcc tagacgtttt tttaacctcg actaaacaca
     5881 tgtaaagcat gtgcaccgag gccccagatc agatcccata caatggggta ccttctgggc
     5941 atccttcagc cccttgttga atacgcttga ggagagccat ttgactcttt ccacaactat
     6001 ccaactcaca acgtggcact ggggttgtgc cgcctttgca ggtgtatctt atacacgtgg
     6061 cttttggccg cagaggcacc tgtcgccagg tggggggttc cgctgcctgc aaagggtcgc
     6121 tacagacgtt gtttgtcttc aagaagcttc cagaggaact gcttccttca cgacattcaa
     6181 cagaccttgc attcctttgg cgagagggga aagaccccta ggaatgctcg tcaagaagac
     6241 agggccaggt ttccgggccc tcacattgcc aaaagacggc aatatggtgg aaaataacat
     6301 atagacaaac gcacaccggc cttattccaa gcggcttcgg ccagtaacgt tagggggggg
     6361 ggagggagag gggcggatcc tcagttactc atgcatacgc gtcatatggg ccgcggatcc
     6421 gatttatcga accactttgt acaagaaagt tgggcatcaa tctggtcgaa aaatcggtgt
     6481 ctttggtaaa aattctatga ggatgacata ttccatcatg ttattcgttt cacatttcct
     6541 tttgctcagt ctccaatgca agcacagctt gtggtataac ctattgtttt cccattctaa
     6601 taaccgctca atcgatcttc gtgttctttt actgaggcaa ataactggcc acatactgca
     6661 acctaatgtg cagcagcaac aaaggcagcc acaaagtagc caacgtacat taacaggaag
     6721 gttcttctta agacaactgt taactctgtt gatgctggct ttaaattctt caggagctac
     6781 ttttccagtt aatgaagaag ggaattcaga ttcaaatttg ttgctcagtc caaatacggt
     6841 gacgtgaccg gagccgcgga cgaccaccgg gtccggcgag tacttgagca gctgctcctc
     6901 cagggcccgc tcctcgtcct cctcttcctc atagatttcg tcgaaatccg cggtgctatc
     6961 caggcccagc agcgggttcg gaatcggttt gcccatgcca acttttttgt acaaacttgt
     7021 ttgataaatt cgaaccggtg gatccgtcga cggtatcgat aagcttgatt cgaccggccg
     7081 actcagtctg tcggaggact ggcgcgccga gtgaggggtt gtgagctctt ttatagagct
     7141 cgggaagcag aagcgcgcga acagaagcga gaagcaggct gattggttaa ttcaaataag
     7201 gcgcagggtc atttcaggtc cttgggggag cctggaaaca tctgatgggt cttaagaaac
     7261 tgctgagggt tgggccatat ctggggacca tctgttcttg gcctcgggcc ggggccgaaa
     7321 ctgcggtgac catctgttct tggccccggg ccggggccga aactgctcac cgcagatatc
     7381 ctgtttggcc caacgttagc tattttcatg tacccgccct tgatctgaac ttctctattc
     7441 ttggtttggt atttttccat gccttgcaaa atggcgttac tgcagctagc tcgaggctag
     7501 tctcgtgatc gataaaattt tgaatttttg taatttgttt ttgtaattct ttagtttgta
     7561 tgtctgttgc tattatgtct actattcttt cccctgcact gtacccccca atcccccctt
     7621 ttcttttaaa agttaaccga taccgtcgag atccgttcac taatcgaatg gatctgtctc
     7681 tgtctctctc tccaccttct tcttctattc cttcgggcct gtcgggtccc ctcggggttg
     7741 ggaggtgggt ctgaaacgat aatggtgaat atccctgcct aactctattc actatagaaa
     7801 gtacagcaaa aactattctt aaacctacca agcctcctac tatcattatg aataatttta
     7861 tataccacag ccaatttgtt atgttaaacc aattccacaa acttgcccat ttatctaatt
     7921 ccaataattc ttgttcattc ttttcttgct ggttttgcga ttcttcaatt aaggagtgta
     7981 ttaagcttgt gtaattgtta atttctctgt cccactccat ccaggtcgtg tgattccaaa
     8041 tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg gtgcaaatga
     8101 gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat ctttccacag
     8161 ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac agatgctgtt
     8221 gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac aataattgtc
     8281 tggcctgtac cgtcagcgtc attgacgctg cgcccatagt gcttcctgct gctcccaaga
     8341 acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc actcttctct
     8401 ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat ttatataatt
     8461 cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagc ggccgcttgc
     8521 tgtgcggtgg tcttactttt gttttgctct tcctcta
//
