LOCUS Exported 4349 bp ds-DNA circular SYN 29-NOV-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS p427-pCSDest-AcrE2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4349) AUTHORS Amrani TITLE Direct Submission JOURNAL Exported Tuesday, Nov 29, 2016 from SnapGene 3.1.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4349 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 35..53 /note="SP6 promoter" /note="promoter for bacteriophage SP6 RNA polymerase" misc_feature 97..105 /note="kozak" misc_feature 106..357 /note="AcrE2" promoter complement(376..393) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 398..532 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" promoter complement(645..663) /note="T3 promoter" /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(684..700) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind 708..724 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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(732..762) /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 777..798 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(1086..1674) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1845..2705) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(2706..2810) /gene="bla" /note="AmpR promoter" rep_origin complement(2836..3291) /direction=LEFT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3365..4349 /note="CMV IE94 promoter" /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" ORIGIN 1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag 61 ctacttgttc tttttgcagg atcccatcga ttcgaagccg ccaccatgaa tacctatctc 121 atcgaccccc gcaaaaacaa cgacaactcc ggcgagcgct tcacggttga cgctgtcgac 181 attacagccg ccgcgaagag cgcagcccaa cagattcttg gcgaggaatt cgagggcctc 241 gtataccgtg aaaccgggga gagtaacgga agtggcatgt tccaggccta ccaccacctg 301 cacggcacta accgcacgga gacgaccgtt ggctatccgt ttcatgtaat ggaactctag 361 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat 421 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat 481 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa 541 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc 601 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta 661 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 721 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 781 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 841 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 901 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 961 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 1021 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 1081 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 1141 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 1201 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 1261 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 1321 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 1381 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 1441 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 1501 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 1561 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 1621 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 1681 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 1741 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 1801 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 1861 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 1921 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 1981 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 2041 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 2101 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 2161 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 2221 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 2281 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 2341 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 2401 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 2461 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 2521 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 2581 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 2641 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 2701 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 2761 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 2821 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt 2881 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa 2941 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa 3001 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac 3061 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga 3121 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa 3181 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc 3241 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc 3301 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc 3361 agtcgaccat agccaattca atatggcgta tatggactca tgccaattca atatggtgga 3421 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga 3481 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga 3541 cttggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg 3601 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat 3661 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg 3721 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg 3781 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg 3841 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct 3901 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt 3961 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc 4021 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac 4081 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc 4141 aagtacatta ctattggaag gacgccaggg tacattggca gtactcccat tgacgtcaat 4201 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg 4261 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg 4321 tctatataag caatgctcgt ttagggaac //