LOCUS Exported 4274 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBbE7k-RFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4274) 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 4274) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4274 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(8..1090) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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" primer_bind 1052..1071 /label=LacI-R /note="LacI, reverse primer" promoter complement(1091..1168) /gene="lacI" /label=lacI promoter primer_bind 1374..1393 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 1481..1500 /label=T7 /note="T7 promoter, forward primer" promoter 1481..1499 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1504..1520 /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)." CDS 1561..2238 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" terminator 2271..2342 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2358..2385 /label=T7Te terminator /note="phage T7 early transcription terminator" rep_origin complement(2543..3131) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2623..2642) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 3219..3313 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3344..4138) /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(3455..3474) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4065..4084 /label=Neo-R /note="Neomycin resistance gene, reverse primer" ORIGIN 1 gacgtcctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 61 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca 121 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca 181 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg 241 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa 301 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa 361 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg 421 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat 481 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca 541 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt 601 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg 661 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa 721 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga 781 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt 841 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa 901 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct 961 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca 1021 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta 1081 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc 1141 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc 1201 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat 1261 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc 1321 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg 1381 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt 1441 ccggcgtaga ggatcgagat cgatctcgat cccgcgaaat taatacgact cactataggg 1501 gaattgtgag cggataacaa tttcagaatt caaaagatct tttaagaagg agatatacat 1561 atggcgagta gcgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt 1621 tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt 1681 acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc 1741 ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc tgacatcccg 1801 gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat gaacttcgaa 1861 gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga gttcatctac 1921 aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca gaaaaaaacc 1981 atgggttggg aagcttccac cgaacgtatg tacccggaag acggtgctct gaaaggtgaa 2041 atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt taaaaccacc 2101 tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat caaactggac 2161 atcacctccc acaacgaaga ctacaccatc gttgaacagt acgaacgtgc tgaaggtcgt 2221 cactccaccg gtgcttaagg atccaaactc gagtaaggat ctccaggcat caaataaaac 2281 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc 2341 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat acctagggcg 2401 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 2461 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 2521 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 2581 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 2641 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 2701 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 2761 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 2821 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 2881 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 2941 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct 3001 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 3061 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 3121 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 3181 actcacgtta agggattttg gtcatgacta gtgcttggat tctcaccaat aaaaaacgcc 3241 cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct 3301 atcaacagga gtccaagcga gctctcgaac cccagagtcc cgctcagaag aactcgtcaa 3361 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga 3421 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt 3481 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat 3541 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt 3601 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt 3661 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc 3721 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca 3781 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct 3841 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca 3901 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca 3961 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg 4021 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga 4081 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc 4141 gaaacgatcc tcatcctgtc tcttgatcag atcatgatcc cctgcgccat cagatccttg 4201 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc 4261 cagctggcaa ttcc //