LOCUS Exported 5238 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRAJ31 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5238) AUTHORS Rodrigo G, Landrain TE, Jaramillo A TITLE De novo automated design of small RNA circuits for engineering synthetic riboregulation in living cells. JOURNAL Proc Natl Acad Sci U S A. 2012 Sep 18;109(38):15271-6. Epub 2012 Sep 4. PUBMED 22949707 REFERENCE 2 (bases 1 to 5238) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5238 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..22 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" CDS complement(26..703) /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" RBS 710..721 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" protein_bind 953..971 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 978..996 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 1014..1028 /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)." protein_bind 1035..1051 /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 1098..1109 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 1125..1835 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" misc_feature 1842..1862 /label=BioBrick suffix /note="universal suffix for all parts" terminator 1898..1929 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" CDS complement(2028..2843) /codon_start=1 /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 2751..2770 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2835..2854 /label=pENTR-R /note="pENTR vectors, reverse primer" CDS complement(3201..4151) /codon_start=1 /gene="rep101" /product="RepA protein needed for replication with the pSC101 origin" /label=Rep101 /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI" rep_origin 4199..4421 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" terminator 4913..5007 /label=lambda t0 terminator /note="transcription terminator from phage lambda" terminator 5163..5192 /label=T3Te terminator /note="phage T3 early transcription terminator" ORIGIN 1 gaattcgcgg ccgcttctag agttattaag caccggtgga gtgacgacct tcagcacgtt 61 cgtactgttc aacgatggtg tagtcttcgt tgtgggaggt gatgtccagt ttgatgtcgg 121 ttttgtaagc acccggcagc tgaaccggtt ttttagccat gtaggtggtt ttaacttcag 181 cgtcgtagtg accaccgtct ttcagtttca gacgcatttt gatttcacct ttcagagcac 241 cgtcttccgg gtacatacgt tcggtggaag cttcccaacc catggttttt ttctgcataa 301 ccggaccgtc ggacgggaag ttggtaccac gcagtttaac tttgtagatg aactcaccgt 361 cttgcaggga ggagtcctgg gtaacggtaa caacaccacc gtcttcgaag ttcataacac 421 gttcccattt gaaaccttcc gggaaggaca gtttcaggta gtccgggatg tcagccgggt 481 gtttaacgta agctttggaa ccgtactgga actgcgggga caggatgtcc caagcgaacg 541 gcagcggacc acctttggta actttcagtt tagcggtctg ggtaccttcg tacggacgac 601 cttcaccttc accttcgatt tcgaactcgt gaccgttaac ggaaccttcc atacgaactt 661 tgaaacgcat gaactctttg ataacgtctt cggaggaagc catctagtat ttctcctctt 721 tctctagtgg atccgctagc atcgatgcga aaaaaccccg ccgaagcggg gttttttgcg 781 gctagcaaaa aaagatcgta gccatgtagc aaatagctac taagaccaat acagtgacag 841 cggataggga cttaattgcg gatgagacga cggtaaagct taatattgat cctagcaacc 901 atttaccaca taactacgac ccaggcgctg gacgtttgat gtgtgctcag tatctctatc 961 actgataggg atgtcaatct ctatcactga tagggaccta ggatgcatat aaatgtgagc 1021 ggataacatt gacattgtga gcggataaca agatactgag cacagtggat atgctttacc 1081 tcttggtgag acgacataaa gaggagaaac ataatatgac tagccgtaaa ggcgaagagc 1141 tgttcactgg tgtcgtccct attctggtgg aactggatgg tgatgtcaac ggtcataagt 1201 tttccgtgcg tggcgagggt gaaggtgacg caactaatgg taaactgacg ctgaagttca 1261 tctgtactac tggtaaactg ccggtacctt ggccgactct ggtaacgacg ctgacttatg 1321 gtgttcagtg ctttgctcgt tatccggacc atatgaagca gcatgacttc ttcaagtccg 1381 ccatgccgga aggctatgtg caggaacgca cgatttcctt taaggatgac ggcacgtaca 1441 aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt aaaccgcatt gagctgaaag 1501 gcattgactt taaagaagac ggcaatatcc tgggccataa gctggaatac aattttaaca 1561 gccacaatgt ttacatcacc gccgataaac aaaaaaatgg cattaaagcg aattttaaaa 1621 ttcgccacaa cgtggaggat ggcagcgtgc agctggctga tcactaccag caaaacactc 1681 caatcggtga tggtcctgtt ctgctgccag acaatcacta tctgagcacg caaagcgttc 1741 tgtctaaaga tccgaacgag aaacgcgatc atatggttct gctggagttc gtaaccgcag 1801 cgggcatcac gcatggtatg gatgaactgt acaaatgatg atactagtag cggccgctgc 1861 aggagtcact aagggttagt tagttagatt agcagaaagt caaaagcctc cgaccggagg 1921 cttttgacta aaacttccct tggggttatc attggggctc actcaaaggc ggtaatcaga 1981 taaaaaaaat ccttagcttt cgctaaggat gatttctgct agtattatta gaaaaactca 2041 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga 2101 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccaaag aatggcaagg 2161 tcctggtaac ggtctgcgat tccgacccgt ccaacatcaa tacaacctat taatttcccc 2221 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag 2281 aatggcaaga gcttgtgcat ttctttccag acttgttcaa caggccagcc attacgctcg 2341 tcatcaaaat cactcgcatc aaccaaaccg ttattcatgc gtgattgcgc ctgagcaaga 2401 cgaaatacac gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg taaccggcgc 2461 aggaacacgg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc 2521 tggaaggctg ttttcccagg aatcgcggtg gtgagtaacc acgcatcatc aggagtacgg 2581 ataaaatgct tgatggtcgg gagaggcata aactccgtca gccagttgag acggaccatc 2641 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca 2701 tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc 2761 catttatacc catataaatc agcgtccatg ttggagttta agcgcggacg ggagcaagac 2821 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt 2881 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac 2941 acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca gctctagtag 3001 ttacattgtc gatctgttca tggtgaacag ctttaaatgc accaaaaact cgtaaaagct 3061 ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg 3121 atatctaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag 3181 atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg 3241 gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatgctt actttgcatg 3301 tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt 3361 agtgtttttc ttagtccgtt acgtaggtag gaatctgatg taatggttgt tggtattttg 3421 tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct 3481 agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc 3541 atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt 3601 ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc 3661 tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc 3721 ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt 3781 tttaactgga aaagataagg caatatctct tcactaaaaa ctaattctaa tttttcgctt 3841 gagaacttgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg 3901 atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt 3961 tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct 4021 ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg 4081 gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact 4141 aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg 4201 gctagtcaat gataattaca tgtccttttc ctttgagttg tgggtatctg taaattctgc 4261 tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct 4321 ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa 4381 aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg 4441 cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac 4501 cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc 4561 tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac 4621 ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag 4681 gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg 4741 tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc 4801 agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta 4861 aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt ggattctcac 4921 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg 4981 tcattactgg atctatcaac aggagtccaa gcgagctcgt aaacttggtc tgacagctct 5041 agctccggca aaaaaacggg caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa 5101 agattacttc gcgtttgcca cctgacgtct aagaaaagga atattcagca atttgcccgt 5161 gccgaagaaa ggcccacccg tgaaggtgag ccagtgagtt gattgctacg taattagtta 5221 gttagccctt agtgactc //