LOCUS Exported 6926 bp ds-DNA circular SYN 09-DEC-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6926) AUTHORS . TITLE Direct Submission JOURNAL Exported Tuesday, Nov 24, 2020 from SnapGene Viewer 5.0.8 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6926 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 142..158 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 182..219 /label=Primer 1 misc_feature 202..1461 /label=salI arm primer_bind complement(1444..1481) /label=Primer 2 CDS 1468..1863 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" CDS 1873..1929 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 1930..1953 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 1963..2283 /codon_start=1 /product="F36V mutant of FK506-binding protein FKBP12" /label=DmrB /note="binds synthetic ligands that do not bind wild-type FKBP" /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" primer_bind 2290..2334 CDS 2290..2316 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 2317..3030 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind 2893..2912 /label=EGFP/EYFP AA 195 5' primer_bind 3047..3084 /label=Primer 3 misc_feature 3067..4338 /label=KpniI arm CDS 3067..3139 /codon_start=1 /translation="PGPPSAKRPKLSSPALSASASDGT" primer_bind complement(4321..4373) /label=Primer 4 promoter complement(4429..4447) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4468..4484) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 4492..4508 /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(4516..4546) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4561..4582 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4870..5458) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5629..6489) /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" promoter complement(6490..6594) /gene="bla" /label=AmpR promoter ORIGIN 1 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc 61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag 121 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc cactagtggc cgcggcggcc 181 gcagcttgcc tgcaggtcga cggcagaggc aagaaacaac agccgccatc ttgtttgtgt 241 gctagcctgt ggggggggtg gggggaggag gagagagagc ggagagggac cgggagagag 301 agagtggggc aagcgggacg cccggctgag tgcgaactgc gcgagcaggg agtgaggagg 361 gtacctggct ggagagcagc ggccgggacc gagacagtgg ttgggggccc cggggcgcaa 421 aggctgaggc gaccctcagt cccctccttg cccgcacaca cccccaccgc agcccgggcc 481 ggcgtcgacg cctaaggggg ggaaaccatt acataagccc gcgccccgcg gcgcctctgc 541 ccgccgtccg aggcggcccc cgagccgagc ccccggcagg cacctggctg ccgggggggt 601 gagggccagg gaggcggccg gactgatttc tcctccccgg gcttgcgagt ggaggcggag 661 aagcgcggcg cgcggagagg ccgcgtcggt gggctcggcg tcagcgcggg cttgcgggcg 721 gacggccctg cttggcccgg cccggccccc ctggcccttc gggccggcga gctccacgct 781 cggcgtcgtc gctgctgcgc ctctgcttgt ccgctgcgcg atgtgaggcg gccccggcgc 841 cagtctggct ctcggctcgg ctgagctcag tgtggccatt aggggccggt gcggcagcgg 901 cggcgcggag cgcagcggca ggaggaggag gagggcggct ctcagggtgg gggcgcaggc 961 ccgggagggg gcaccgggag gaggtgagtg tctctggtcg cctcctcctc tccccctttt 1021 tcgcccccgc ctccttgtgg cgatgagaag gaggaggaca gcgccgagga ggaagaagct 1081 gatggcggcg gcggagctcc gagagacctc ggctgggcag gggctcgccg tggcgggctg 1141 gggactgcgt ctgtagagct gtgagttccc gagaattcgc tgcagcggac gtgctcgtgg 1201 gatcagtgtt gctgtaccct cctcccggcc tggtttaggc ccggcccctc gcacttgccc 1261 ctaccatttc tgttgagtcc gcatccctct ccggccaccg caaccaggcg aagagaaaaa 1321 ggaacttccc cccacccctg ttgggtggcg gcggagctcc ggagcccacc cttaggagcg 1381 gggcggggac cccgtttcgg agaagagatt tcctcaggat tctcgttttt ttcctcggtt 1441 gtatctccga aagaattaaa agtcgacatg gccaagcctt tgtctcaaga agaatccacc 1501 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc 1561 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt 1621 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc 1681 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga 1741 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt 1801 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag 1861 ggcggttccg gagccacgaa cttctctctg ttaaagcaag caggagacgt ggaagaaaac 1921 cccggtcccg attacaagga tgacgatgac aagggttccg gcggcgtgca ggtggagact 1981 atctccccag gagacgggcg caccttcccc aagcgcggcc agacctgcgt ggtgcactac 2041 accgggatgc ttgaagatgg aaagaaagtt gattcctccc gggacagaaa caagcccttt 2101 aagtttatgc taggcaagca ggaggtgatc cgaggctggg aagaaggggt tgcccagatg 2161 agtgtgggtc agagagccaa actgactata tctccagatt atgcctatgg tgccactggg 2221 cacccaggca tcatcccacc acatgccact ctcgtcttcg atgtggagct tctaaaactg 2281 gaagtcgatt atccgtacga cgtaccagac tacgcagtga gcaagggcga ggagctgttc 2341 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc 2401 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc 2461 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcctg 2521 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg 2581 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc 2641 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc 2701 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac 2761 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc 2821 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc 2881 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc 2941 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 3001 atcactctcg gcatggacga gctgtacaag ggtggaggtg gatctggtgg cggcggttca 3061 ggtacccccg ggccgccttc agccaagcgg cctaaactct catctccggc cctctcggcg 3121 tccgccagcg atggcacagg ttagtctcgg gactcccggc cttccatagt tcccttcgtc 3181 ttttctgctc gttgcacctt cattccaggg tgcttctgtg tatctcttcc catttccccg 3241 gccggtgttt aattttaaag gtttttgaat ggactggtcg aagacgtcca gtagcccaac 3301 cattttcttt gccttctgat acattcattg caacactatg tcatgtcaga tcgttgtgtg 3361 ttctggaatt aagaggttgg aagtgggagc tgcggggacc tttattgttg gctccatgca 3421 atttaatttg ggaaatgccg gtacaatcgt gtgtgagggc ttttatgcac ttttactatt 3481 tttgttactg gtgaatgagg gatttcagct ctttctcccc tgttgcaatg aattcttttt 3541 ttctctgctt tcccaccttt aatgattttt gctcctgtga ggcagatttt ctgttgagga 3601 atgggggcgg aagacaaatc ttggtgtctg cgatccaagg ttttaagtgt aactaaagtt 3661 caacaaaatt tcattcttag gtcccctctc cccatccttg cctcaagagt cagtatggac 3721 cgcagtttat tttatatcca tttgccatga aaccaaaatg tctcaaactg cgtgttgagg 3781 tgtagacctg aggcgtttcc tttgcagttt gtcacactct tacttagaag ggaaagaaaa 3841 aagatgaagc ctttctggaa ttaggtactt agaaatggga gataataagt tgaaagactc 3901 ccagtcatga acttccaact aaagtgattg tacaagttta aaaatgaaca ggaggttttg 3961 tgcttgcatg tttgttttgt tttgcttttt ggacctttgc aattccagtt tacaagatga 4021 aggaattctt ttctccagtc agcccctttg ttaggagacc tgaagggaga ctctcccttg 4081 tctgttcttt ttcctgctta ctgttagttt agtgttattc tgatgatcac tgacctggtc 4141 ctctcttgct gtctgacttg tttgagaggc caggataatg aagaagaaga agaagaaaaa 4201 gaagaagaag aagaagaatg gctagtcatc tgacagtcca ctagatgttc ctccttactg 4261 caagcatccc tttaactcag agaagaggct ccagatctct ccttctgcca atgagagtgt 4321 atagtggagt taaacgtagg taccgggccc cccctcgagg tcgatcgacg gtatcgataa 4381 gcttgatcta gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg 4441 gttaattccg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 4501 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 4561 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 4621 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 4681 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 4741 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 4801 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 4861 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 4921 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 4981 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 5041 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 5101 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 5161 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 5221 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 5281 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 5341 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 5401 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 5461 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5521 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 5581 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 5641 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 5701 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 5761 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 5821 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 5881 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 5941 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6001 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6061 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6121 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6181 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6241 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 6301 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6361 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6421 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6481 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6541 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6601 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa 6661 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct 6721 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag 6781 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 6841 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg 6901 cgtaaggaga aaataccgca tcaggc //