LOCUS       sequence-11428-e        8246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8246)
  AUTHORS   Wilson AA, Kwok LW, Hovav AH, Ohle SJ, Little FF, Fine A, Kotton DN
  TITLE     Sustained expression of alpha1-antitrypsin after transplantation of 
            manipulated hematopoietic stem cells.
  JOURNAL   Am J Respir Cell Mol Biol. 2008 Aug . 39(2):133-41.
  PUBMED    18323534
REFERENCE   2  (bases 1 to 8246)
  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: "Am J Respir
            Cell Mol Biol. 2008 Aug . 39(2):133-41."
COMMENT     Sequence Label: pHAGE-CMV-dsRed-UBC-GFP-W
FEATURES             Location/Qualifiers
     source          1..8246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2227..2530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2531..2734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2685..2705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2731..2755
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2806..2815
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2812..3489
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(2942..2960)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(2992..3012)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3210..3229
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3404..3427
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     promoter        3496..3894
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             join(3903..3905,3906..3908,3909..4622)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3903..3905,3906..3908,3909..4619)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3948..3969)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4209..4228)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4556..4577
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4636..5224
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4689..4709)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5107..5118)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5299..5532
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5651..5669)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5737..5841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5842..5910,5911..6702)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6060..6079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6873..7461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7362..7381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7615..7632
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7872..7893
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7908..7913,7914..7931,7932..7938)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7946..7962
                     /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     7951..7973
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7970..7986
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7970..7986
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8036..8055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttcacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagtgga
     2221 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     2281 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     2341 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     2401 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     2461 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     2521 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     2581 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     2641 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     2701 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     2761 gagacgccat ccacgctgtt ttgacctcca tagaagacac cggcggccgc catggcctcc
     2821 tccgaggacg tcatcaagga gttcatgcgc ttcaaggtgc gcatggaggg ctccgtgaac
     2881 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     2941 gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat cctgtccccc
     3001 cagttccagt acggctccaa ggtgtacgtg aagcaccccg ccgacatccc cgactacaag
     3061 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     3121 gtggtgaccg tgacccagga ctcctccctg caggacggct ccttcatcta caaggtgaag
     3181 ttcatcggcg tgaacttccc ctccgacggc cccgtaatgc agaagaagac tatgggctgg
     3241 gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga gatccacaag
     3301 gccctgaagc tgaaggacgg cggccactac ctggtggagt tcaagtccat ctacatggcc
     3361 aagaagcccg tgcagctgcc cggctactac tacgtggact ccaagctgga catcacctcc
     3421 cacaacgagg actacaccat cgtggagcag tacgagcgcg ccgagggccg ccaccacctg
     3481 ttcctgtagg gatccggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcc
     3541 tcacggcgag cgctgccacg tcagacgaag ggcgcagcga gcgtcctgat ccttccgccc
     3601 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     3661 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     3721 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     3781 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     3841 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggccgccc
     3901 atatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     3961 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     4021 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     4081 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     4141 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4201 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4261 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4321 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     4381 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     4441 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4501 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     4561 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     4621 aaatcgatag atcctaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     4681 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     4741 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     4801 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     4861 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4921 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4981 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     5041 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     5101 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     5161 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     5221 ccgcctgaga tcctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     5281 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     5341 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     5401 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     5461 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     5521 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     5581 aatgaatatc agagagtgag aggcccgggt taattaagga aagggctaga tcattcttga
     5641 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5701 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5821 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5881 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5941 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6001 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6061 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     6121 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6181 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6241 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6301 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6361 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6421 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6481 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6541 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6601 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6661 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6721 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6781 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6841 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6901 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6961 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7021 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7081 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7141 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7201 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7261 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7321 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7381 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7441 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7501 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7561 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7621 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7681 ccgattcatt aatgcagcaa gctcatggct gactaatttt ttttatttat gcagaggccg
     7741 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     7801 gcttttgcaa aaagctcccc gtggcacgac aggtttcccg actggaaagc gggcagtgag
     7861 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7921 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7981 tatgacatga ttacgaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     8041 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
     8101 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
     8161 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
     8221 taaatatgta ctacaaactt agtagt
//
