LOCUS Exported 4860 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pSTC1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4860) 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 4860) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4860 /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)" CDS 747..1457 /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 1464..1484 /label=BioBrick suffix /note="universal suffix for all parts" terminator 1520..1551 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" CDS complement(1650..2465) /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 2373..2392 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2457..2476 /label=pENTR-R /note="pENTR vectors, reverse primer" CDS complement(2823..3773) /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 3821..4043 /label=pSC101 ori /note="low-copy replication origin that requires the Rep101 protein" terminator 4535..4629 /label=lambda t0 terminator /note="transcription terminator from phage lambda" terminator 4785..4814 /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 atcccctagg gctagccgta aaggcgaaga gctgttcact ggtgtcgtcc 781 ctattctggt ggaactggat ggtgatgtca acggtcataa gttttccgtg cgtggcgagg 841 gtgaaggtga cgcaactaat ggtaaactga cgctgaagtt catctgtact actggtaaac 901 tgccggtacc ttggccgact ctggtaacga cgctgactta tggtgttcag tgctttgctc 961 gttatccgga ccatatgaag cagcatgact tcttcaagtc cgccatgccg gaaggctatg 1021 tgcaggaacg cacgatttcc tttaaggatg acggcacgta caaaacgcgt gcggaagtga 1081 aatttgaagg cgataccctg gtaaaccgca ttgagctgaa aggcattgac tttaaagaag 1141 acggcaatat cctgggccat aagctggaat acaattttaa cagccacaat gtttacatca 1201 ccgccgataa acaaaaaaat ggcattaaag cgaattttaa aattcgccac aacgtggagg 1261 atggcagcgt gcagctggct gatcactacc agcaaaacac tccaatcggt gatggtcctg 1321 ttctgctgcc agacaatcac tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg 1381 agaaacgcga tcatatggtt ctgctggagt tcgtaaccgc agcgggcatc acgcatggta 1441 tggatgaact gtacaaatga tgatactagt agcggccgct gcaggagtca ctaagggtta 1501 gttagttaga ttagcagaaa gtcaaaagcc tccgaccgga ggcttttgac taaaacttcc 1561 cttggggtta tcattggggc tcactcaaag gcggtaatca gataaaaaaa atccttagct 1621 ttcgctaagg atgatttctg ctagtattat tagaaaaact catcgagcat caaatgaaac 1681 tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat 1741 gaaggagaaa actcaccgag gcagttccaa agaatggcaa ggtcctggta acggtctgcg 1801 attccgaccc gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta 1861 tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa gagcttgtgc 1921 atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca 1981 tcaaccaaac cgttattcat gcgtgattgc gcctgagcaa gacgaaatac acgatcgctg 2041 ttaaaaggac aattacaaac aggaatcgaa tgtaaccggc gcaggaacac ggccagcgca 2101 tcaacaatat tttcacctga atcaggatat tcttctaata cctggaaggc tgttttccca 2161 ggaatcgcgg tggtgagtaa ccacgcatca tcaggagtac ggataaaatg cttgatggtc 2221 gggagaggca taaactccgt cagccagttg agacggacca tctcatctgt aacatcattg 2281 gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaag 2341 cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa 2401 tcagcgtcca tgttggagtt taagcgcgga cgggagcaag acgtttcccg ttgaatatgg 2461 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat 2521 atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttgttga 2581 ataaatcgaa cttttgctga gttgaaggat cagctctagt agttacattg tcgatctgtt 2641 catggtgaac agctttaaat gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt 2701 ttacaccgtt ttcatctgtg catatggaca gttttccctt tgatatctaa cggtgaacag 2761 ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga gccataagaa 2821 cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg tttttgcgtg 2881 agccatgaga acgaaccatt gagatcatgc ttactttgca tgtcactcaa aaattttgcc 2941 tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg 3001 ttacgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt catttttatc 3061 tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac ttggaaaatc 3121 aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct gtaagtgttt 3181 aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc atggtagtta 3241 ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt tgccttgtga 3301 gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga gtatttgttt 3361 tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg gaaaagataa 3421 ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt ggcatagttt 3481 gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac agttctcgtc 3541 atcagctctc tggttgcttt agctaataca ccataagcat tttccctact gatgttcatc 3601 atctgagcgt attggttata agtgaacgat accgtccgtt ctttccttgt agggttttca 3661 atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg ctccgttaag 3721 tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga catacatctc 3781 aattggtcta ggtgatttta atcactatac caattgagat gggctagtca atgataatta 3841 catgtccttt tcctttgagt tgtgggtatc tgtaaattct gctagacctt tgctggaaaa 3901 cttgtaaatt ctgctagacc ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt 3961 tatattcaag tggttataat ttatagaata aagaaagaat aaaaaaagat aaaaagaata 4021 gatcccagcc ctgtgtataa ctcactactt tagtcagttc cgcagtatta caaaaggatg 4081 tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa accctaaagg cttaagtagc 4141 accctcgcaa gctcgggcaa atcgctgaat attccttttg tctccgacca tcaggcacct 4201 gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc acggctctgg cagtgaatgg 4261 gggtaaatgg cactacaggc gccttttatg gattcatgca aggaaactac ccataataca 4321 agaaaagccc gtcacgggct tctcagggcg ttttatggcg ggtctgctat gtggtgctat 4381 ctgacttttt gctgttcagc agttcctgcc ctctgatttt ccagtctgac cacttcggat 4441 tatcccgtga caggtcattc agactggcta atgcacccag taaggcagcg gtatcatcaa 4501 caggcttacc cgtcttactg tccctagtgc ttggattctc accaataaaa aacgcccggc 4561 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca 4621 acaggagtcc aagcgagctc gtaaacttgg tctgacagct ctagctccgg caaaaaaacg 4681 ggcaaggtgt caccaccctg ccctttttct ttaaaaccga aaagattact tcgcgtttgc 4741 cacctgacgt ctaagaaaag gaatattcag caatttgccc gtgccgaaga aaggcccacc 4801 cgtgaaggtg agccagtgag ttgattgcta cgtaattagt tagttagccc ttagtgactc //