LOCUS Exported 3642 bp ds-DNA circular SYN 11-DEC-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3642) AUTHORS Molecular Function Laboratory TITLE Direct Submission JOURNAL Exported Dec 11, 2018 from SnapGene Viewer 4.2.9 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3642 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 4..459 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 701..706 /label=Kozak seq CDS 707..1102 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" CDS 1115..1171 /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 1178..1381 /codon_start=1 /label=mAID /note="note=""direct strand translation""translation=""KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNV MVSCQKSSGGPEAAAFVKVSMDGAPYLRKIDLRMYK""" /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA AAFVKVSMDGAPYLRKIDLRMYK" CDS 1382..1399 /codon_start=1 /label=Linker /translation="GAGAGA" CDS 1388..1399 /codon_start=1 /label=Linker-mAID-Clover /translation="GAGA" misc_feature 1388..1397 /label=(965)_pAID_Linker_5'_BamHI_SacI promoter complement(1454..1472) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1493..1509) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 1517..1533 /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(1541..1571) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1586..1607 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(1895..2483) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2654..3514) /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(3515..3619) /gene="bla" /label=AmpR promoter ORIGIN 1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg 481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg 661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gccaccatgg ccaagccttt 721 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat 781 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg 841 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc 901 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg 961 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa 1021 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc 1081 ctctggttat gtgtgggagg gctcaggaag cggagctact aacttcagcc tgctgaagca 1141 ggctggagac gtggaggaga accctggacc tagatctaag gagaagagtg cttgtcctaa 1201 agatccagcc aaacctccgg ccaaggcaca agttgtggga tggccaccgg tgagatcata 1261 ccggaagaac gtgatggttt cctgccaaaa atcaagcggt ggcccggagg cggcggcgtt 1321 cgtgaaggta tcaatggacg gagcaccgta cttgaggaaa atcgatttga ggatgtataa 1381 aggcgctggt gcaggcgccg gatccactag ttctagagcg gccgccaccg cggtggagct 1441 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc 1501 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 1561 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 1621 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 1681 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 1741 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 1801 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 1861 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 1921 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 1981 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 2041 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 2101 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 2161 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 2221 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 2281 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 2341 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 2401 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 2461 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 2521 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 2581 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 2641 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 2701 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 2761 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 2821 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 2881 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 2941 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 3001 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 3061 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 3121 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 3181 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 3241 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 3301 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 3361 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 3421 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 3481 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 3541 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 3601 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc ac //