LOCUS       sequence-73358-e        8270 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AND gate in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8270)
  AUTHORS   Stanton BC, Nielsen AA, Tamsir A, Clancy K, Peterson T, Voigt CA
  TITLE     Genomic mining of prokaryotic repressors for orthogonal logic gates.
  JOURNAL   Nat Chem Biol. 2013 Dec 8. doi: 10.1038/nchembio.1411.
  PUBMED    24316737
REFERENCE   2  (bases 1 to 8270)
  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: "Nat Chem 
            Biol. 2013 Dec 8. doi: 10.1038/nchembio.1411."
COMMENT     Sequence Label: pAND
FEATURES             Location/Qualifiers
     source          1..8270
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(284..828)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(364..383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(917..1732)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(917..1732)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1640..1659
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1724..1743
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     terminator      2634..2705
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2721..2748
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    2779..2797
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2804..2822
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      3460..3531
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3547..3574
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        join(3587..3592,3593..3608,3609..3615)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    3621..3640
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     terminator      4300..4371
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4387..4414
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             4496..4507
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(4514..4516,4517..4519,4520..5233)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4559..4580)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4820..4839)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5167..5188
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      5257..5328
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5344..5371
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             5464..6546
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5483..5502)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             7357..7980
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(7430..7449)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(8163..8182)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 cacgaggcag acctcagcgc tagcggagtg tatactggct tactatgttg gcactgatga
       61 gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag
      121 aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
      181 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
      241 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
      301 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
      361 tataaagata ccaggcgttt cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt
      421 tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac
      481 tcagttccgg gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt
      541 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa
      601 aagcaccact ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc
      661 gccggttaag gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta
      721 cctcggttca aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt
      781 ttttcgtttt cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt
      841 attaaggggt ctgacgctca gtggaacgaa aaatcaatct aaagtatata tgagtaaact
      901 tggtctgaca gttaccttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
      961 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     1021 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     1081 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     1141 caccatgagt gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga
     1201 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     1261 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     1321 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     1381 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     1441 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     1501 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     1561 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     1621 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     1681 tggaatttaa tcgcggcctc gagcaagacg tttcccgttg aatatggctc ataacacccc
     1741 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     1801 gtgcaatgta catcagagat tttgagacac aaggtctccc gctcgacgta cggtggaatc
     1861 tgattcgtta ccaattgaca tgatacgaaa cgtaccgtat cgttaaggtg gtatggaagc
     1921 tatacgttac caattgacag ctagctcagt cctactttag tatatagacc gtgcgatcgg
     1981 tctatagcta cgacttgctc atttgacaga ggataactac tagtgccgaa actgggtatg
     2041 cagagcattc gtcgtcgtca gctgattgat gcaaccctgg aagcaattaa tgaagttggt
     2101 atgcatgatg caaccattgc acagattgca cgtcgtgccg gtgttagcac cggtattatt
     2161 agccattatt tccgcgataa aaacggtctg ctggaagcaa ccatgcgtga tattaccagc
     2221 cagctgcgtg atgcagttct gaatcgtctg catgcactgc cgcagggtag cgcagaacag
     2281 cgtctgcagg caattgttgg tggtaatttt gatgaaaccc aggttagcag cgcagcaatg
     2341 aaagcatggc tggcattttg ggcaagcagc atgcatcagc cgatgctgta tcgtctgcag
     2401 caggttagca gtcgtcgtct gctgagcaat ctggttagcg aatttcgtcg tgaactgcct
     2461 cgtgaacagg cacaagaggc aggttatggt ctggcagcac tgattgatgg tctgtggctg
     2521 cgtgcagcac tgagcggtaa accgctggat aaaacccgtg caaatagcct gacccgtcat
     2581 tttatcaccc agcatctgcc gaccgattaa tcgtaaggcg agcccccagg catcaaataa
     2641 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2701 ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatagcttta
     2761 ctccaccgtt ggcttttttc cctatcagtg atagagattg acatccctat cagtgataga
     2821 gataatgagc acgccatgcc attggctttt gatagaggac aactactaga tgaacctgaa
     2881 agataaaatt ctgggcgttg ccaaagaact gtttatcaaa aatggctata acgcaaccac
     2941 caccggtgaa attgttaaac tgagcgaaag cagcaaaggc aatctgtatt atcactttaa
     3001 aaccaaagag aacctgtttc tggaaatcct gaacatcgaa gaaagcaaat ggcaagagca
     3061 gtggaaaaaa gaacaaatca aatgcaaaac caaccgcgag aaattctatc tgtataatga
     3121 actgagcctg accaccgaat attactatcc gctgcagaat gccatcatcg agttttatac
     3181 cgagtactat aaaaccaaca gcatcaacga gaaaatgaac aaactggaaa acaaatacat
     3241 cgatgcctac cacgtgatct ttaaagaagg taatctgaac ggcgaatggt gcattaatga
     3301 tgttaatgcc gtgagcaaaa ttgcagcaaa tgccgttaat ggcattgtta cctttaccca
     3361 tgagcagaat atcaacgaac gcattaaact gatgaacaaa ttcagccaga tctttctgaa
     3421 tggcctgagc aaataaggac tgcgacgttc tccaggcatc aaataaaacg aaaggctcag
     3481 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     3541 cacactggct caccttcggg tgggcctttc tgcgtttata cgcttgttga caattaatca
     3601 tcggctcgta taatgtgtgg aattgtgagc gctcacaatt ctatggacta tgtttgaaag
     3661 ggagaaatac tagatggcac gtaccccgag ccgtagcagc attggtagcc tgcgtagtcc
     3721 gcatacccat aaagcaattc tgaccagcac cattgaaatc ctgaaagaat gtggttatag
     3781 cggtctgagc attgaaagcg ttgcacgtcg tgccggtgca agcaaaccga ccatttatcg
     3841 ttggtggacc aataaagcag cactgattgc cgaagtgtat gaaaatgaaa gcgaacaggt
     3901 gcgtaaattt ccggatctgg gtagctttaa agccgatctg gattttctgc tgcgtaatct
     3961 gtggaaagtt tggcgtgaaa ccatttgtgg tgaagcattt cgttgtgtta ttgcagaagc
     4021 acagctggac cctgcaaccc tgacccagct gaaagatcag tttatggaac gtcgtcgtga
     4081 gatgccgaaa aaactggttg aaaatgccat tagcaatggt gaactgccga aagataccaa
     4141 tcgtgaactg ctgctggata tgatttttgg tttttgttgg tatcgcctgc tgaccgaaca
     4201 gctgaccgtt gaacaggata ttgaagaatt taccttcctg ctgattaatg gtgtttgtcc
     4261 gggtacacag cgttaactag ggcccatacc cccaggcatc aaataaaacg aaaggctcag
     4321 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     4381 cacactggct caccttcggg tgggcctttc tgcgtttata ggagagcgcg ggtgagaggg
     4441 attcgttacc aattgacaat tgattggacg ttcaatataa tgctagctac tagagaaaga
     4501 ggagaaatac tagatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     4561 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     4621 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     4681 gccctggccc accctcgtga ccaccttcgg ctacggcctg caatgcttcg cccgctaccc
     4741 cgaccacatg aagctgcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     4801 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     4861 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     4921 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     4981 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     5041 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     5101 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     5161 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     5221 gctgtacaag taactagggc ccataccccc aggcatcaaa taaaacgaaa ggctcagtcg
     5281 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
     5341 actggctcac cttcgggtgg gcctttctgc gtttataaat ggcggcgcgc catcgaatgg
     5401 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     5461 aatatgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     5521 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     5581 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     5641 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     5701 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     5761 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     5821 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     5881 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     5941 attttctccc atgaggacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     6001 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     6061 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     6121 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     6181 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     6241 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagatagc
     6301 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     6361 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     6421 ccagtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     6481 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6541 cagtgataag atcctattcc agcgggtcac acaggaaagg cctcgatgaa aaacataaat
     6601 gccgacgaca catacagaat aattaataaa attaaagctt gtagaagcaa taatgatatt
     6661 aatcaatgct tatctgatat gactaaaatg gtacattgtg aatattattt actcgcgatc
     6721 atttatcctc attctatggt taaatctgat atttcaatcc tagataatta ccctaaaaaa
     6781 tggaggcaat attatgatga cgctaattta ataaaatatg atcctatagt agattattct
     6841 aactccaatc attcaccaat taattggaat atatttgaaa acaatgctgt aaataaaaaa
     6901 tctccaaatg taattaaaga agcgaaaaca tcaggtctta tcactgggtt tagtttccct
     6961 attcatacgg ctaacaatgg cttcggaatg cttagttttg cacattcaga aaaagacaac
     7021 tatatagata gtttattttt acatgcgtgt atgaacatac cattaattgt tccttctcta
     7081 gttgataatt atcgaaaaat aaatatagca aataataaat caaacaacga tttaaccaaa
     7141 agagaaaaag aatgtttagc gtgggcatgc gaaggaaaaa gctcttggga tatttcaaaa
     7201 atattaggtt gcagtgagcg tactgtcact ttccatttaa ccaatgcgca aatgaaactc
     7261 aatacaacaa accgctgcca aagtatttct aaagcaattt taacaggagc aattgattgc
     7321 ccatacttta aaaattaagg atccaggagg aaaaaaatgt ccagattaga taaaagtaaa
     7381 gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt
     7441 aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt aaaaaataag
     7501 cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac tcacttttgc
     7561 cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag ttttagatgt
     7621 gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc tacagaaaaa
     7681 cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt ttcactagag
     7741 aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt attggaagat
     7801 caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag tatgccgcca
     7861 ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc cttcttattc
     7921 ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag tgggtcctaa
     7981 taattggtaa cgaatcagac aattgacggc tcgagggagt agcatagggt ttgcagaatc
     8041 cctgcttcgt ccatttgaca ggcacattat gcatcgatga taagctgtca aacatgagca
     8101 gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     8161 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     8221 caaatatttt atctgaggtc gagaccgctt cctcgctcac tgactcgctg
//
