LOCUS Exported 5153 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRAJ12 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5153) 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 5153) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5153 /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 868..886 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 893..911 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 929..943 /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 950..966 /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 1013..1024 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 1040..1750 /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 1757..1777 /label=BioBrick suffix /note="universal suffix for all parts" terminator 1813..1844 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" CDS complement(1943..2758) /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 2666..2685 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2750..2769 /label=pENTR-R /note="pENTR vectors, reverse primer" CDS complement(3116..4066) /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 4114..4336 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" terminator 4828..4922 /label=lambda t0 terminator /note="transcription terminator from phage lambda" terminator 5078..5107 /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 gctagcccca gaatcccttc gagcaaatcc tagcgaggtg ccgctagatt cgctcaaagg 841 aacccttctt cctgcccgtg ctcagtatct ctatcactga tagggatgtc aatctctatc 901 actgataggg acctaggatg catataaatg tgagcggata acattgacat tgtgagcgga 961 taacaagata ctgagcacac ccagtatcat tctcttcttc ctgcccacgc ggaaagagga 1021 gaaaggtgta atgactagcc gtaaaggcga agagctgttc actggtgtcg tccctattct 1081 ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg agggtgaagg 1141 tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta aactgccggt 1201 accttggccg actctggtaa cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc 1261 ggaccatatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct atgtgcagga 1321 acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag tgaaatttga 1381 aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag aagacggcaa 1441 tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca tcaccgccga 1501 taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg aggatggcag 1561 cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc ctgttctgct 1621 gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga acgagaaacg 1681 cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg gtatggatga 1741 actgtacaaa tgatgatact agtagcggcc gctgcaggag tcactaaggg ttagttagtt 1801 agattagcag aaagtcaaaa gcctccgacc ggaggctttt gactaaaact tcccttgggg 1861 ttatcattgg ggctcactca aaggcggtaa tcagataaaa aaaatcctta gctttcgcta 1921 aggatgattt ctgctagtat tattagaaaa actcatcgag catcaaatga aactgcaatt 1981 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag 2041 aaaactcacc gaggcagttc caaagaatgg caaggtcctg gtaacggtct gcgattccga 2101 cccgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg 2161 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caagagcttg tgcatttctt 2221 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca 2281 aaccgttatt catgcgtgat tgcgcctgag caagacgaaa tacacgatcg ctgttaaaag 2341 gacaattaca aacaggaatc gaatgtaacc ggcgcaggaa cacggccagc gcatcaacaa 2401 tattttcacc tgaatcagga tattcttcta atacctggaa ggctgttttc ccaggaatcg 2461 cggtggtgag taaccacgca tcatcaggag tacggataaa atgcttgatg gtcgggagag 2521 gcataaactc cgtcagccag ttgagacgga ccatctcatc tgtaacatca ttggcaacgc 2581 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aagcgataga 2641 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcgt 2701 ccatgttgga gtttaagcgc ggacgggagc aagacgtttc ccgttgaata tggctcataa 2761 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt 2821 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttgt tgaataaatc 2881 gaacttttgc tgagttgaag gatcagctct agtagttaca ttgtcgatct gttcatggtg 2941 aacagcttta aatgcaccaa aaactcgtaa aagctctgat gtatctatct tttttacacc 3001 gttttcatct gtgcatatgg acagttttcc ctttgatatc taacggtgaa cagttgttct 3061 acttttgttt gttagtcttg atgcttcact gatagataca agagccataa gaacctcaga 3121 tccttccgta tttagccagt atgttctcta gtgtggttcg ttgtttttgc gtgagccatg 3181 agaacgaacc attgagatca tgcttacttt gcatgtcact caaaaatttt gcctcaaaac 3241 tggtgagctg aatttttgca gttaaagcat cgtgtagtgt ttttcttagt ccgttacgta 3301 ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc attcattttt atctggttgt 3361 tctcaagttc ggttacgaga tccatttgtc tatctagttc aacttggaaa atcaacgtat 3421 cagtcgggcg gcctcgctta tcaaccacca atttcatatt gctgtaagtg tttaaatctt 3481 tacttattgg tttcaaaacc cattggttaa gccttttaaa ctcatggtag ttattttcaa 3541 gcattaacat gaacttaaat tcatcaaggc taatctctat atttgccttg tgagttttct 3601 tttgtgttag ttcttttaat aaccactcat aaatcctcat agagtatttg ttttcaaaag 3661 acttaacatg ttccagatta tattttatga atttttttaa ctggaaaaga taaggcaata 3721 tctcttcact aaaaactaat tctaattttt cgcttgagaa cttggcatag tttgtccact 3781 ggaaaatctc aaagccttta accaaaggat tcctgatttc cacagttctc gtcatcagct 3841 ctctggttgc tttagctaat acaccataag cattttccct actgatgttc atcatctgag 3901 cgtattggtt ataagtgaac gataccgtcc gttctttcct tgtagggttt tcaatcgtgg 3961 ggttgagtag tgccacacag cataaaatta gcttggtttc atgctccgtt aagtcatagc 4021 gactaatcgc tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt 4081 ctaggtgatt ttaatcacta taccaattga gatgggctag tcaatgataa ttacatgtcc 4141 ttttcctttg agttgtgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa 4201 attctgctag accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc 4261 aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca 4321 gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa 4381 cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg 4441 caagctcggg caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc 4501 tgtctttttc gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa 4561 tggcactaca ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag 4621 cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc tatctgactt 4681 tttgctgttc agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg 4741 tgacaggtca ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaggctt 4801 acccgtctta ctgtccctag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc 4861 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag 4921 tccaagcgag ctcgtaaact tggtctgaca gctctagctc cggcaaaaaa acgggcaagg 4981 tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt tgccacctga 5041 cgtctaagaa aaggaatatt cagcaatttg cccgtgccga agaaaggccc acccgtgaag 5101 gtgagccagt gagttgattg ctacgtaatt agttagttag cccttagtga ctc //