LOCUS Exported 3442 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBbA2k-RFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3442) AUTHORS Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, Keasling JD TITLE BglBrick vectors and datasheets: A synthetic biology platform for gene expression. JOURNAL J Biol Eng. 2011 Sep 20;5:12. PUBMED 21933410 REFERENCE 2 (bases 1 to 3442) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3442 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(7..630) /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" primer_bind 538..557 /label=Tet-R /note="Tetracycline resistance gene, reverse primer" promoter 649..704 /gene="tetR" /label=tetR/tetA promoters /note="overlapping promoters for bacterial tetR and tetA" protein_bind 685..703 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" CDS 742..1419 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" terminator 1452..1523 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1539..1566 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(1594..1611) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1728..2273) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(1808..1827) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 2387..2481 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(2512..3306) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2623..2642) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 3233..3252 /label=Neo-R /note="Neomycin resistance gene, reverse primer" ORIGIN 1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc 61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg 121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg 181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa 241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc 301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag 361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc 421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa 481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc 541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt 601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta 661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcaaaagatc 721 ttttaagaag gagatataca tatggcgagt agcgaagacg ttatcaaaga gttcatgcgt 781 ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt 841 gaaggtcgtc cgtacgaagg tacccagacc gctaaactga aagttaccaa aggtggtccg 901 ctgccgttcg cttgggacat cctgtccccg cagttccagt acggttccaa agcttacgtt 961 aaacacccgg ctgacatccc ggactacctg aaactgtcct tcccggaagg tttcaaatgg 1021 gaacgtgtta tgaacttcga agacggtggt gttgttaccg ttacccagga ctcctccctg 1081 caagacggtg agttcatcta caaagttaaa ctgcgtggta ccaacttccc gtccgacggt 1141 ccggttatgc agaaaaaaac catgggttgg gaagcttcca ccgaacgtat gtacccggaa 1201 gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac 1261 gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac 1321 aaaaccgaca tcaaactgga catcacctcc cacaacgaag actacaccat cgttgaacag 1381 tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaag gatccaaact cgagtaagga 1441 tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct 1501 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt 1561 tctgcgttta tacctaggga tatattccgc ttcctcgctc actgactcgc tacgctcggt 1621 cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc 1681 caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc 1741 cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca 1801 ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct 1861 gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct 1921 gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt 1981 tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca 2041 tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt 2101 catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc 2161 agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg 2221 cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc 2281 atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag 2341 cacctgaagt cagccccata cgatataagt tgttactagt gcttggattc tcaccaataa 2401 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta 2461 ctggatctat caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa 2521 ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag 2581 cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa 2641 cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa 2701 gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc 2761 ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg 2821 atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg 2881 ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag 2941 ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag 3001 gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac 3061 gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc 3121 gtcctgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc 3181 ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc 3241 atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc 3301 aatcatgcga aacgatcctc atcctgtctc ttgatcagat catgatcccc tgcgccatca 3361 gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga 3421 gggcgcccca gctggcaatt cc //