LOCUS pETDuet-1 5420 bp ds-DNA DEFINITION synthetic circular DNA KEYWORDS pETDuet-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5420) AUTHORS HonorGene: http://www.honorgene.com FEATURES Location/Qualifiers source 1..5420 /organism="synthetic DNA construct" /mol_type="other DNA" source 42..64 /organism="Enterobacteria phage T7" /mol_type="genomic DNA" /db_xref="taxon:10760" protein_bind 3..27 /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)." RBS 42..64 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 83..100 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" promoter 214..232 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 233..257 /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)." CDS 366..410 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /note="S-Tag" /translation="KETAAAKFERQHMDS" terminator 462..509 /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 546..1001 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(1116..1976) /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(1977..2069) /gene="bla" /note="AmpR promoter" rep_origin 2150..2738 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 2924..3066 /note="bom" /note="basis of mobility region from pBR322" CDS complement(3168..3359) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /note="rop" /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(3931..5013) /codon_start=1 /gene="lacI" /product="lac repressor" /note="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" promoter complement(5014..5091) /gene="lacI" /note="lacI promoter" promoter 5404..2 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag 61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag 121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag 181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt 241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata 301 tggcagatct caattggata tcggccggcc acgcgatcgc tgacgtcggt accctcgagt 361 ctggtaaaga aaccgctgct gcgaaatttg aacgccagca catggactcg tctactagcg 421 cagcttaatt aacctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg 481 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga 541 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt 601 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct 661 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg 721 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag 781 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa 841 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga 901 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa 961 atttaacgcg aattttaaca aaatattaac gtttacaatt tctggcggca cgatggcatg 1021 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 1081 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 1141 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 1201 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 1261 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 1321 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 1381 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 1441 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 1501 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 1561 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 1621 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 1681 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 1741 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 1801 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 1861 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 1921 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 1981 ttcctttttc aatcatgatt gaagcattta tcagggttat tgtctcatga gcggatacat 2041 atttgaatgt atttagaaaa ataaacaaat aggtcatgac caaaatccct taacgtgagt 2101 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 2161 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 2221 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 2281 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 2341 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 2401 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 2461 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 2521 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 2581 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 2641 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 2701 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 2761 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 2821 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 2881 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc 2941 tccttacgca tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg 3001 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg 3061 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg 3121 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca 3181 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc 3241 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt 3301 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact 3361 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag 3421 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag 3481 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc 3541 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg 3601 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca 3661 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg 3721 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag 3781 cctagccggg tcctcaacga caggagcacg atcatgctag tcatgccccg cgcccaccgg 3841 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga 3901 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 3961 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 4021 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc 4081 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc 4141 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc 4201 cactaccgag atgtccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc 4261 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat 4321 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg 4381 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac 4441 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc 4501 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc 4561 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc 4621 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg 4681 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc 4741 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc 4801 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac 4861 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc 4921 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata 4981 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc 5041 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc 5101 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt 5161 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg 5221 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag 5281 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc 5341 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatcgatctc gatcccgcga 5401 aattaatacg actcactata //