LOCUS Exported 4164 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRAJ11m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4164) 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 4164) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4164 /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""" protein_bind 33..47 /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 54..70 /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 117..128 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 147..857 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI CTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV TAAGITHGMDELYK" primer_bind complement(171..199) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 794..814 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 877..948 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 964..991 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_feature 998..1018 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(1102..1119) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1273..1861) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1353..1372) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2094..2909) /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 2817..2836 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2901..2920 /label=pENTR-R /note="pENTR vectors, reverse primer" CDS complement(3064..3924) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3687..3706 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3925..4029) /gene="bla" /label=AmpR promoter primer_bind 4097..4115 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 4119..4162 /label=bacterial terminator /note="putative bacterial transcription terminator" ORIGIN 1 gaattcgcgg ccgcttctag agctagcata aatgtgagcg gataacattg acattgtgag 61 cggataacaa gatactgagc accctcgcat aatttcactt cttcaatcct cccgttaaag 121 aggagaaatt atgaatgact agccgtaaag gagaagaact tttcactgga gttgtcccaa 181 ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg 241 aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac 301 ctgttccatg gccaacactt gtcactactt tcggttatgg tgttcaatgc tttgcgagat 361 acccagatca tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac 421 aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt 481 ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg 541 gaaacattct tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg 601 cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac attgaagatg 661 gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc 721 ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa 781 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg 841 atgaactata caaataataa tactagagcc aggcatcaaa taaaacgaaa ggctcagtcg 901 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac 961 actggctcac cttcgggtgg gcctttctgc gtttatatac tagtagcggc cgctgcagtc 1021 cggcaaaaaa acgggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt 1081 acttcgcgtt atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 1141 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 1201 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 1261 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 1321 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 1381 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 1441 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 1501 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 1561 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 1621 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 1681 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct 1741 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 1801 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 1861 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 1921 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 1981 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gctcgagtcc 2041 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa 2101 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat 2161 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg 2221 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat 2281 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc 2341 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta 2401 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga 2461 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac 2521 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct 2581 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga 2641 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg 2701 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct 2761 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg 2821 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag 2881 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca 2941 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt 3001 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagctc 3061 gagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 3121 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 3181 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 3241 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 3301 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 3361 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 3421 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 3481 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 3541 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 3601 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 3661 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 3721 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 3781 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 3841 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 3901 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 3961 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 4021 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat 4081 gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt 4141 agctttcgct aaggatgatt tctg //