LOCUS Exported 3741 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For course in ""Synthetic Biology: A Lab Manual"". ACCESSION . VERSION . KEYWORDS pSB3K3 containing mRFP1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3741) AUTHORS Liljeruhm J, Funk SK, Tietscher S, Edlund AD, Jamal S, Wistrand-Yuen P, Dyrhage K, Gynna A, Ivermark K, Lovgren J, Tornblom V, Virtanen A, Lundin ER, Wistrand-Yuen E, Forster AC TITLE Engineering a palette of eukaryotic chromoproteins for bacterial synthetic biology. JOURNAL J Biol Eng. 2018 May 10;12:8. doi: 10.1186/s13036-018-0100-0. eCollection 2018. PUBMED 29760772 REFERENCE 2 (bases 1 to 3741) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3741 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 17..60 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 63..84 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" protein_bind 185..206 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 221..251 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 259..275 /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)." RBS 293..304 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 311..988 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" terminator 1033..1104 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1120..1147 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_feature 1154..1174 /label=BioBrick suffix /note="universal suffix for all parts" CDS complement(1314..2129) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 2037..2056 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2121..2140 /label=pENTR-R /note="pENTR vectors, reverse primer" rep_origin complement(2724..3268) /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(2803..2822) /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3736..13 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc 61 tggaattcgc ggccgcttct agagcaatac gcaaaccgcc tctccccgcg cgttggccga 121 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg 181 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg 241 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacatactag agaaagagga 301 gaaatactag atggcttcct ccgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg 361 tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc 421 gtacgaaggt acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc 481 ttgggacatc ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc 541 tgacatcccg gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat 601 gaacttcgaa gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga 661 gttcatctac aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca 721 gaaaaaaacc atgggttggg aagcttccac cgaacgtatg tacccggaag acggtgctct 781 gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt 841 taaaaccacc tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat 901 caaactggac atcacctccc acaacgaaga ctacaccatc gttgaacagt acgaacgtgc 961 tgaaggtcgt cactccaccg gtgcttaata acgctgatag tgctagtgta gatcgctact 1021 agagccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc 1081 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct 1141 ttctgcgttt atatactagt agcggccgct gcaggcttcc tcgctcactg actcgctgcg 1201 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tctcgagtcc 1261 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa 1321 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc gataccatat 1381 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg 1441 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat 1501 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc 1561 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta 1621 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga 1681 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac 1741 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct 1801 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga 1861 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg 1921 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct 1981 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg 2041 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag 2101 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca 2161 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt 2221 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagatc 2281 acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac 2341 tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg gctggatgat 2401 ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag acctcagcgc 2461 tagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc 2521 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata 2581 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat 2641 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt 2701 gagagggccg cggcaaagcc gtttttccat aggctccgcc ccctgacaag catcacgaaa 2761 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc 2821 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc 2881 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt 2941 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc 3001 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc 3061 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga 3121 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt 3181 agctcagaga accttcaaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga 3241 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaaggggt ctgacgctca 3301 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 3361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 3421 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 3481 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 3541 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 3601 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 3661 cgcctccatc cagtctattc catggtgcca cctgacgtct aagaaaccat tattatcatg 3721 acattaacct ataaaaatag g //