LOCUS Exported 3273 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION For course in ""Synthetic Biology: A Lab Manual"". ACCESSION . VERSION . KEYWORDS pSB1K3 containing mRFP1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3273) 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 3273) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3273 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 15..45 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 53..69 /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 87..98 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 105..782 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" terminator 827..898 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 914..941 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_feature 948..968 /label=BioBrick suffix /note="universal suffix for all parts" terminator 969..1026 /label=his operon terminator /note="This putative transcriptin terminator from the E. coli his operon has a 2-bp deletion introduced during synthesis. Its efficiency has not been determined." primer_bind complement(1050..1067) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1221..1809) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1301..1320) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2042..2857) /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 2765..2784 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2849..2868 /label=pENTR-R /note="pENTR vectors, reverse primer" primer_bind 3062..3080 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 3084..3127 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 3130..3151 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" protein_bind 3252..3273 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc 61 ggataacaat ttcacacata ctagagaaag aggagaaata ctagatggct tcctccgaag 121 acgttatcaa agagttcatg cgtttcaaag ttcgtatgga aggttccgtt aacggtcacg 181 agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga aggtacccag accgctaaac 241 tgaaagttac caaaggtggt ccgctgccgt tcgcttggga catcctgtcc ccgcagttcc 301 agtacggttc caaagcttac gttaaacacc cggctgacat cccggactac ctgaaactgt 361 ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt cgaagacggt ggtgttgtta 421 ccgttaccca ggactcctcc ctgcaagacg gtgagttcat ctacaaagtt aaactgcgtg 481 gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa aaccatgggt tgggaagctt 541 ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg tgaaatcaaa atgcgtctga 601 aactgaaaga cggtggtcac tacgacgctg aagttaaaac cacctacatg gctaaaaaac 661 cggttcagct gccgggtgct tacaaaaccg acatcaaact ggacatcacc tcccacaacg 721 aagactacac catcgttgaa cagtacgaac gtgctgaagg tcgtcactcc accggtgctt 781 aataacgctg atagtgctag tgtagatcgc tactagagcc aggcatcaaa taaaacgaaa 841 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta 901 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatatac tagtagcggc 961 cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc tttaaaaccg 1021 aaaagattac ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt 1081 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 1141 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 1201 aaggccgcgt tgctggcgtt tttccacagg ctccgccccc ctgacgagca tcacaaaaat 1261 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 1321 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 1381 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 1441 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 1501 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 1561 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 1621 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 1681 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 1741 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 1801 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 1861 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 1921 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc 1981 tcgagtcccg tcaagtcagc gtaatgctct gccagtgtta caaccaatta accaattctg 2041 attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa 2101 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc 2161 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac 2221 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga 2281 ctgaatccgg tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc 2341 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt 2401 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg 2461 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat 2521 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat 2581 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt 2641 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa 2701 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga 2761 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg 2821 gcctggagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta 2881 tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc 2941 agagattttg agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg 3001 atcagctcga gtgccacttg acgtctaaga aaccattatt atcatgacat taacctataa 3061 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga 3121 tgatttctgg aattcgcggc cgcttctaga gcaatacgca aaccgcctct ccccgcgcgt 3181 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag 3241 cgcaacgcaa ttaatgtgag ttagctcact cat //