LOCUS       40924_43253        3721 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTHSSe_36, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3721)
  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 3721)
  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 3721)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3721)
  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..3721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..22)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     terminator      25..68
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     rep_origin      558..1102
                     /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."
     CDS             1697..2485
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     terminator      complement(2646..2703)
                     /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."
     misc_feature    complement(2704..2724)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      complement(2731..2758)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2774..2845)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2865..3578)
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     regulatory      complement(3579..3597)
                     /note="B0032; derived from BBa_B0032"
                     /regulatory_class="ribosome_binding_site"
     misc_RNA        complement(3621..3671)
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      complement(3673..3721)
                     /label=PPhlF
                     /note="PPhlF"
                     /regulatory_class="promoter"
     protein_bind    complement(3673..3703)
                     /label=PhlF binding site
                     /bound_moiety="PhlF"
ORIGIN
        1 ctctagaagc ggccgcgaat tccagaaatc atccttagcg aaagctaagg atttttttta
       61 tctgaaattc tgcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
      121 ttcttagacg tcaggtggca ccatggaata gactggatgg aggcggataa agttgcagga
      181 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
      241 gagcgtgggt atcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
      301 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
      361 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
      421 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
      481 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
      541 ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac
      601 gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac
      661 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
      721 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
      781 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      841 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
      901 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
      961 accgaaaggc aggaacagga gagcgcacga gggagccgcc aggggaaacg cctggtatct
     1021 ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc
     1081 aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt
     1141 atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc
     1201 agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac
     1261 atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga
     1321 caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgct gaggtctgcc
     1381 tcgtgaagaa ggtgttgctg actcatacca ggcctgaatc gccccatcat ccagccagaa
     1441 agtgagggag ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa
     1501 cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa
     1561 ctcagcaaaa gttcgattta ttcaacaaag ccacgttgtg tctcaaaatc tctgatgtta
     1621 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     1681 taatacaagg ggtgttatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga
     1741 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
     1801 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
     1861 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
     1921 ttcccctgta gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
     1981 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
     2041 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
     2101 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
     2161 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
     2221 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
     2281 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
     2341 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
     2401 agaagatcgc ttggcctcgc gcacagatca gttggaagaa tttgtccact acgtgaaagg
     2461 cgagatcacc aaggtagtcg gcaaataatc agaattggtt aattggttgt aacactggca
     2521 gagcattacg ctgacttgac gggactcgag attaccgcct ttgagtgagc tgataccgct
     2581 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcctg cataacgcga
     2641 agtaatcttt tcggttttaa agaaaaaggg cagggtggtg acaccttgcc cttttttgcc
     2701 ggactgcagc ggccgctact agtatataaa cgcagaaagg cccacccgaa ggtgagccag
     2761 tgtgactcta gtagagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     2821 tcgactgagc ctttcgtttt atttgatgcc tggctctagt attatttgta gagctcatcc
     2881 atgccatgtg taatcccagc agcagttaca aactcaagaa ggaccatgtg gtcacgcttt
     2941 tcgttgggat ctttcgaaag gacagattgt gtcgacaggt aatggttgtc tggtaaaagg
     3001 acagggccat cgccaattgg agtattttgt tgataatggt ctgctagttg aacggaacca
     3061 tcttcaacgt tgtggcgaat tttgaagtta gctttgattc cattcttttg tttgtctgcc
     3121 gtgatgtata cattgtgtga gttaaagttg tactcgagtt tgtgtccaag aatgtttcca
     3181 tcttctttaa aatcaatacc ctttaactcg atacgattaa caagggtatc accttcaaac
     3241 ttgacttcag cacgcgtctt gtaggtcccg tcatctttga aagatatagt gcgttcctgt
     3301 acataacctt cgggcatggc actcttgaaa aagtcatgcc gtttcatgtg atccggataa
     3361 cgggaaaagc attgaacacc ataggtcaga gtagtgacaa gtgttggcca cggaacaggt
     3421 agttttccag tagtgcaaat aaatttaagg gtgagttttc cgtttgtagc atcaccttca
     3481 ccctctccac ggacagaaaa tttgtgccca ttaacatcac catctaattc aacaagaatt
     3541 gggacaactc cagtgaaaag ttcttctcct ttgctcatct agtactttcc tgtgtgatta
     3601 aacaaaatta tttgtagagg ctgtttcgtc ctcacggact catcagaccg gaaagcacat
     3661 ccggtgacag ctaaccttaa cgatacggta cgtttcgtat catgtcaatt ggtaacgaat
     3721 c
//
