LOCUS Exported 3176 bp ds-DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pTHSSe_53, complete sequence. ACCESSION MH029770 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3176) AUTHORS Segall-Shapiro TH, Sontag ED, Voigt CA. TITLE Engineered promoters enable constant gene expression at any copy number in bacteria JOURNAL Nat. Biotechnol. (2018) In press PUBMED 29553576 REFERENCE 2 (bases 1 to 3176) AUTHORS Segall-Shapiro TH, Sontag ED, Voigt CA. TITLE Direct Submission JOURNAL Submitted (06-MAR-2018) Synthetic Biology Center, Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA REFERENCE 3 (bases 1 to 3176) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..3176 /organism="Cloning vector pTHSSe_53" /mol_type="other DNA" /label=pUC BbsI cloning vector /note="pUC BbsI cloning vector" /db_xref="taxon:2137934" regulatory 13..60 /regulatory_class="promoter" /note="PJ23105; derived from BBa_J23105" ncRNA 61..135 /product="RiboJ" /label=ribozyme /note="insulator" /ncRNA_class="ribozyme" misc_RNA 63..112 /label=sTRSV HHRz /note="hammerhead ribozyme from the tobacco ringspot virus satellite RNA (Khvorova et al., 2003)" regulatory 136..154 /regulatory_class="ribosome_binding_site" /note="B0032; derived from BBa_B0032" CDS 155..871 /codon_start=1 /product="sfGFP" /label=sfGFP /protein_id="AVR55154.1" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" CDS 155..868 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" regulatory 880..1008 /regulatory_class="terminator" /note="B0015; BBa_B0015" terminator 888..959 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 975..1002 /label=T7Te terminator /note="phage T7 early transcription terminator" misc_feature 1021..1041 /label=BioBrick suffix /note="universal suffix for all parts" terminator 1042..1099 /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." rep_origin 1195..1958 /label=pUC origin /note="pUC origin" rep_origin 1271..1859 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2054..2914) /codon_start=1 /product="TEM beta-lactamase" /label=TEM beta-lactamase /note="ampicillin resistance protein" /protein_id="AVR55153.1" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(2915..3019) /gene="bla" /label=AmpR promoter terminator 3109..3152 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 3155..3176 /label=BioBrick prefix /note="BioBrick prefix for parts that do not start with ""ATG""" ORIGIN 1 ggaggtgtct tcggcgcgcc tttacggcta gctcagtcct aggtactatg ctagcaaggt 61 agctgtcacc ggatgtgctt tccggtctga tgagtccgtg aggacgaaac agcctctaca 121 aataattttg tttaatcaca caggaaagta ctagatgagc aaaggagaag aacttttcac 181 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt 241 ccgtggagag ggtgaaggtg atgctacaaa cggaaaactc acccttaaat ttatttgcac 301 tactggaaaa ctacctgttc cgtggccaac acttgtcact actctgacct atggtgttca 361 atgcttttcc cgttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc 421 cgaaggttat gtacaggaac gcactatatc tttcaaagat gacgggacct acaagacgcg 481 tgctgaagtc aagtttgaag gtgataccct tgttaatcgt atcgagttaa agggtattga 541 ttttaaagaa gatggaaaca ttcttggaca caaactcgag tacaacttta actcacacaa 601 tgtatacatc acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca 661 caacgttgaa gatggttccg ttcaactagc agaccattat caacaaaata ctccaattgg 721 cgatggccct gtccttttac cagacaacca ttacctgtcg acacaatctg tcctttcgaa 781 agatcccaac gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat 841 tacacatggc atggatgagc tctacaaata atactagagc caggcatcaa ataaaacgaa 901 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct 961 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttataga agacatcgct 1021 tactagtagc ggccgctgca gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt 1081 ttctttaaaa ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca ctgactcgct 1141 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taattcatga 1201 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 1261 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 1321 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 1381 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag 1441 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 1501 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 1561 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 1621 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 1681 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 1741 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 1801 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 1861 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 1921 tctttcctgc gttatcccct gattctgtgg ataaccgtcg attatcaaaa aggatcttca 1981 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 2041 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 2101 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 2161 taccatctgg ccccagtgct gcaatgatac cgcgagaacc acgctcaccg gctccagatt 2221 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 2281 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 2341 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 2401 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 2461 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 2521 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 2581 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 2641 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 2701 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 2761 cgctgttgag atccagttcg atataaccca ctcgtgcacc caactgatct tcagcatctt 2821 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 2881 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 2941 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 3001 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca 3061 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa 3121 aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt ctagag //