LOCUS Exported 8249 bp ds-DNA circular SYN 30-APR-2019 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 8249) AUTHORS timothyaballo TITLE Direct Submission JOURNAL Exported Wednesday, Apr 22, 2020 from SnapGene Viewer 5.0.7 https://www.snapgene.com FEATURES Location/Qualifiers source 1..8249 /organism="recombinant plasmid" /mol_type="other DNA" promoter 351..1529 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" /note="color: #ffffff; direction: RIGHT" intron 582..1520 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" /note="color: #------" primer_bind 1481..1501 /label=Upstrm pEF6 Myc-His Seq /note="color: black; sequence: GCCTCAGACAGTGGTTCAAAG" promoter 1546..1564 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff; direction: RIGHT" primer_bind 1626..1649 /label=Lck Forward (EcoRI) /note="color: black; sequence: TATATGAATtcgccatgggctgtggctgc" CDS 1635..3161 /codon_start=1 /label=Lck /note="color: #ccffcc" /translation="MGCGCSSHPEDDWMENIDVCENCHYPIVPLDGKGTLLIRNGSEVR DPLVTYEGSNPPASPLQDNLVIALHSYEPSHDGDLGFEKGEQLRILEQSGEWWKAQSLT TGQEGFIPFNFVAKANSLEPEPWFFKNLSRKDAERQLLAPGNTHGSFLIRESESTAGSF SLSVRDFDQNQGEVVKHYKIRNLDNGGFYISPRITFPGLHELVRHYTNASDGLCTRLSR PCQTQKPQKPWWEDEWEVPRETLKLVERLGAGQFGEVWMGYYNGHTKVAVKSLKQGSMS PDAFLAEANLMKQLQHQRLVRLYAVVTQEPIYIITEYMENGSLVDFLKTPSGIKLTINK LLDMAAQIAEGMAFIEERNYIHRDLRAANILVSDTLSCKIADFGLARLIEDNEYTAREG AKFPIKWTAPEAINYGTFTIKSDVWSFGILLTEIVTHGRIPYPGMTNPEVIQNLERGYR MVRPDNCPEELYQLMRLCWKERPEDRPTFDYLRSVLEDFFTATEGQYQPQP" primer_bind 2112..2131 /label=LGB-S1N1 /note="color: black; sequence: GCGGGATCGTTTTCACTGTC" primer_bind 2774..2793 /label=LGB-S2N1 /note="color: black; sequence: TGCAGACTTTGGCCTAGCAC" primer_bind complement(3144..3161) /label=Lck Reverse (HA, STOP, PmeI) /note="color: black; sequence: ATATGTTTAAACTCAAGCGTAATCTGGAACATCGTATGGGTAaggctgaggctggtact g" primer_bind 3153..3191 /label=TurboID Forward (BbvCI, Pro, Gly) /note="color: black; sequence: TATATCCTCAGCCTGGAgctagcaaagacaatactgtgcctctg" CDS 3162..3164 /codon_start=1 /label=Gly /note="color: #ff0000" /translation="G" CDS 3165..4133 /codon_start=1 /label=TurboID /note="color: #99cc00" /translation="ASKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLR DWGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKS GDACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRK LGADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQG WITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIG DKEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEKLQ" primer_bind 3516..3535 /label=Turbo Seq Forward /note="color: black; sequence: ggaagcagagggaggaaatg" primer_bind complement(3597..3621) /label=TurboID M157T Reverse (Set 1) /note="color: black; sequence: tgaccgggcccaggccgattgctgc; added: 2019-04-30; 5' phosphorylated" primer_bind complement(3622..3647) /label=TurboID M157T Reverse (Set 2) /note="color: black; sequence: cagcgcttctgccgtgacaattccga; added: 2019-03-14; 5' phosphorylated" primer_bind 3622..3645 /label=TurboID M157T Forward (Set 1) /note="color: black; sequence: tcggaattgtcacggcagaagcgc; added: 2019-04-30; 5' phosphorylated" primer_bind 3648..3672 /label=TurboID M157T Forward (Set 2) /note="color: black; sequence: cgaaagctgggagcagacaaggtgc; added: 2019-03-14; 5' phosphorylated" primer_bind complement(3792..3804) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" /note="color: black; sequence: ATAATACCGCGCCACATAGC" CDS 4134..4160 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /note="color: #993366" /translation="LPPLERLTL" primer_bind complement(4145..4204) /label=Turbo Reverse (Gly, HA, STOP, PmeI) /note="color: black; sequence: TATATGTTTAAACTTAAGCGTAATCTGGAACATCGTATGGGTATCCgtccagggtcagg cgctcc" CDS 4167..4193 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /note="color: #cc99b2" /translation="YPYDVPDYA" polyA_signal 4222..4446 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" /note="color: #a6acb3" primer_bind complement(4351..4371) /label=Dnstrm pEF6 Myc-His Seq /note="color: black; sequence: CACCCCCCAGAATAGAATGAC" rep_origin 4492..4920 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /note="color: #ffff00" promoter 4934..5263 /label=SV40 promoter /note="SV40 enhancer and early promoter" /note="color: #ffffff; direction: RIGHT" rep_origin 5114..5249 /label=SV40 ori /note="SV40 origin of replication" /note="color: #ffff00" promoter 5311..5358 /label=EM7 promoter /note="synthetic bacterial promoter " /note="color: #ffffff; direction: RIGHT" CDS 5377..5775 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=Blasticidin R /note="confers resistance to blasticidin" /note="color: #ccffcc" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 5933..6054 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" /note="color: #a6acb3" primer_bind complement(6103..6119) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: LEFT" protein_bind 6127..6143 /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)." /note="color: #31849b" promoter complement(6151..6181) /label=lac promoter /note="promoter for the E. coli lac operon" /note="This reverse directional feature has 3 segments: 1: 6151 .. 6157 / #ffffff / -10 2: 6158 .. 6175 / #ffffff 3: 6176 .. 6181 / #ffffff / -35" protein_bind 6196..6217 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /note="color: #31849b" rep_origin complement(6494..7082) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(7253..8113) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 7253 .. 8044 / #ccffcc 2: 8045 .. 8113 / #ccffcc / signal sequence Cleavage site after base 8044" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7876..7895 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" /note="color: black; sequence: ATAATACCGCGCCACATAGC" promoter complement(8114..8218) /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: LEFT" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac taggcttttg caaaaagctt tgcaaagatg gataaagttt taaacagaga 301 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg 361 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg 421 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt 481 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg 541 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt 601 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct 661 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga 721 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct 781 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt 841 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc 901 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg 961 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac 1021 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc 1081 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag 1141 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct 1201 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg 1261 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga 1321 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc 1381 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg 1441 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa 1501 gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata cgactcacta 1561 tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac tagtccagtg 1621 tggtggaatt cgccatgggc tgtggctgca gctcacaccc ggaagatgac tggatggaaa 1681 acatcgatgt gtgtgagaac tgccattatc ccatagtccc actggatggc aagggcacgc 1741 tgctcatccg aaatggctct gaggtgcggg acccactggt tacctacgaa ggctccaatc 1801 cgccggcttc cccactgcaa gacaacctgg ttatcgctct gcacagctat gagccctctc 1861 acgacggaga tctgggcttt gagaaggggg aacagctccg catcctggag cagagcggcg 1921 agtggtggaa ggcgcagtcc ctgaccacgg gccaggaagg cttcatcccc ttcaattttg 1981 tggccaaagc gaacagcctg gagcccgaac cctggttctt caagaacctg agccgcaagg 2041 acgcggagcg gcagctcctg gcgcccggga acactcacgg ctccttcctc atccgggaga 2101 gcgagagcac cgcgggatcg ttttcactgt cggtccggga cttcgaccag aaccagggag 2161 aggtggtgaa acattacaag atccgtaatc tggacaacgg tggcttctac atctcccctc 2221 gaatcacttt tcccggcctg catgaactgg tccgccatta caccaatgct tcagatgggc 2281 tgtgcacacg gttgagccgc ccctgccaga cccagaagcc ccagaagccg tggtgggagg 2341 acgagtggga ggttcccagg gagacgctga agctggtgga gcggctgggg gctggacagt 2401 tcggggaggt gtggatgggg tactacaacg ggcacacgaa ggtggcggtg aagagcctga 2461 agcagggcag catgtccccg gacgccttcc tggccgaggc caacctcatg aagcagctgc 2521 aacaccagcg gctggttcgg ctctacgctg tggtcaccca ggagcccatc tacatcatca 2581 ctgaatacat ggagaatggg agtctagtgg attttctcaa gaccccttca ggcatcaagt 2641 tgaccatcaa caaactcctg gacatggcag cccaaattgc agaaggcatg gcattcattg 2701 aagagcggaa ttatattcat cgtgaccttc gggctgccaa cattctggtg tctgacaccc 2761 tgagctgcaa gattgcagac tttggcctag cacgcctcat tgaggacaac gagtacacag 2821 ccagggaggg ggccaagttt cccattaagt ggacagcgcc agaagccatt aactacggga 2881 cattcaccat caagtcagat gtgtggtctt ttgggatcct gctgacggaa attgtcaccc 2941 acggccgcat cccttaccca gggatgacca acccggaggt gattcagaac ctggagcgag 3001 gctaccgcat ggtgcgccct gacaactgtc cagaggagct gtaccaactc atgaggctgt 3061 gctggaagga gcgcccagag gaccggccca cctttgacta cctgcgcagt gtgctggagg 3121 acttcttcac ggccacagag ggccagtacc agcctcagcc tggagctagc aaagacaata 3181 ctgtgcctct gaagctgatc gctctcctgg ctaatggcga gttccatagt ggcgaacagc 3241 tgggagaaac cctgggcatg tccagggccg ctatcaacaa gcacattcag actctgcgcg 3301 actggggcgt ggacgtgttc accgtgcccg gaaagggcta ctctctgccc gagcctatcc 3361 cgctgctgaa cgctaaacag attctgggac agctggacgg cgggagcgtg gcagtcctgc 3421 ctgtggtcga ctccaccaat cagtacctgc tggatcgaat cggcgagctg aagagtgggg 3481 atgcttgcat tgcagaatat cagcaggcag ggagaggaag cagagggagg aaatggttct 3541 ctccttttgg agctaacctg tacctgagta tgttttggcg cctgaagcgg ggaccagcag 3601 caatcggcct gggcccggtc atcggaattg tcatggcaga agcgctgcga aagctgggag 3661 cagacaaggt gcgagtcaaa tggcccaatg acctgtatct gcaggataga aagctggcag 3721 gcatcctggt ggagctggcc ggaataacag gcgatgctgc acagatcgtc attggcgccg 3781 ggattaacgt ggctatgagg cgcgtggagg aaagcgtggt caatcagggc tggatcacac 3841 tgcaggaagc agggattaac ctggacagga atactctggc cgctacgctg atccgagagc 3901 tgcgggcagc cctggaactg ttcgagcagg aaggcctggc tccatatctg ccacggtggg 3961 agaagctgga taacttcatc aatagacccg tgaagctgat cattggggac aaagagattt 4021 tcgggattag ccgggggatt gataaacagg gagccctgct gctggaacag gacggagtta 4081 tcaaaccctg gatgggcgga gaaatcagtc tgcggtctgc cgaaaagctg cagctgcctc 4141 ccctggagcg cctgaccctg gacggatacc catacgatgt tccagattac gcttaagttt 4201 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 4261 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 4321 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 4381 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 4441 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct 4501 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 4561 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 4621 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 4681 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 4741 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 4801 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 4861 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 4921 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 4981 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 5041 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 5101 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 5161 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 5221 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 5281 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc 5341 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt gtctcaagaa 5401 gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac 5461 tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta 5521 tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg 5581 gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc 5641 ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg 5701 aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat 5761 gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt 5821 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 5881 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 5941 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 6001 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 6061 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 6121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 6181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 6241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 6301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 6361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 6421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc caggaaccgt aaaaaggccg 6481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 6541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 6601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 6661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 6721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 6781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 6841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 6901 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 6961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 7021 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 7081 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 7141 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 7201 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 7261 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 7321 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 7381 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 7441 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 7501 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 7561 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 7621 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 7681 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 7741 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 7801 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 7861 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 7921 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 7981 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 8041 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 8101 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 8161 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 8221 catttccccg aaaagtgcca cctgacgtc //