LOCUS       40924_43318        3268 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTHSSe_58, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3268)
  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 3268)
  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 3268)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3268)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Biotechnol. (2018) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAR-2018) Synthetic Biology Center, Department of Biological 
            Engineering, Massachusetts Institute of Technology, 77 Massachusetts
            Avenue, Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      complement(13..102)
                     /label=ECK120029600
                     /note="ECK120029600"
                     /regulatory_class="terminator"
     regulatory      107..175
                     /label=PT7A1w1
                     /note="PT7A1w1"
                     /regulatory_class="promoter"
     misc_RNA        177..227
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      251..269
                     /note="B0032; derived from BBa_B0032"
                     /regulatory_class="ribosome_binding_site"
     CDS             270..983
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     terminator      1003..1074
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1090..1117
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1136..1156
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1157..1214
                     /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      1327..1872
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     terminator      complement(2243..2337)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2361..3017)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3018..3121)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(3201..3244)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3247..3268
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
ORIGIN
        1 gaagacacgg agttcagcca aaaaacttaa gaccgccggt cttgtccact accttgcagt
       61 aatgcggtgg acaggatcgg cggttttctt ttctcttctc aagcttggcg cgcctcagtc
      121 agagtattga cttaaagtct aacctatagg acctttacag ccatcgagag ctgcgagctg
      181 tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct ctacaaataa
      241 ttttgtttaa tcacacagga aagtactaga tgagcaaagg agaagaactt ttcactggag
      301 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtccgtg
      361 gagagggtga aggtgatgct acaaacggaa aactcaccct taaatttatt tgcactactg
      421 gaaaactacc tgttccgtgg ccaacacttg tcactactct gacctatggt gttcaatgct
      481 tttcccgtta tccggatcac atgaaacggc atgacttttt caagagtgcc atgcccgaag
      541 gttatgtaca ggaacgcact atatctttca aagatgacgg gacctacaag acgcgtgctg
      601 aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaagggt attgatttta
      661 aagaagatgg aaacattctt ggacacaaac tcgagtacaa ctttaactca cacaatgtat
      721 acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt cgccacaacg
      781 ttgaagatgg ttccgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg
      841 gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgtcctt tcgaaagatc
      901 ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct gggattacac
      961 atggcatgga tgagctctac aaataatact agagccaggc atcaaataaa acgaaaggct
     1021 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag
     1081 agtcacactg gctcaccttc gggtgggcct ttctgcgttt atacgctatg tcttctacta
     1141 gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc cctttttctt
     1201 taaaaccgaa aagattactt cgcgttatgc aggcttcctc gctcactgac tcgctgcgct
     1261 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaatt taataagatg
     1321 atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc
     1381 cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg
     1441 gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga
     1501 cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat
     1561 gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa
     1621 cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca
     1681 ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca
     1741 ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct
     1801 gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg
     1861 agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg
     1921 catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga
     1981 ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct
     2041 gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc
     2101 agtgccaaca tagtaagcca gtatacactc cgctacgatt atcaaaaagg atcttcacct
     2161 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     2221 ggtctgacag ctcgaggctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     2281 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     2341 gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     2401 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     2461 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     2521 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     2581 gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     2641 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     2701 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     2761 tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc
     2821 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     2881 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     2941 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     3001 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     3061 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc
     3121 aactcgagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3181 taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga
     3241 tttctggaat tcgcggccgc ttctagag
//
