LOCUS       40924_43333        3092 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTHSSe_68, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3092)
  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 3092)
  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 3092)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3092)
  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..3092
                     /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..272
                     /note="RBS32B; derived from BBa_B0032"
                     /regulatory_class="ribosome_binding_site"
     CDS             273..875
                     /codon_start=1
                     /product="PhlF"
                     /label=PhlF
                     /protein_id="AVR55191.1"
                     /translation="MARTPSRSSIGSLRSPHTHKAILTSTIEILKECGYSGLSIESVAR
                     RAGASKPTIYRWWTNKAALIAEVYENESEQVRKFPDLGSFKADLDFLLRNLWKVWRETI
                     CGEAFRCVIAEAQLDPATLTQLKDQFMERRREMPKKLVENAISNGELPKDTNRELLLDM
                     IFGFCWYRLLTEQLTVEQDIEEFTFLLINGVCPGTQR"
     terminator      889..960
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      976..1003
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1022..1042
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1043..1100
                     /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      1272..1860
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2067..2161)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2185..2841)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(2842..2945)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(3025..3068)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    3071..3092
                     /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 tactcacaca ggaaagtact agatggcacg taccccgagc cgtagcagca
      301 ttggtagcct gcgtagtccg catacccata aagcaattct gaccagcacc attgaaatcc
      361 tgaaagaatg tggttatagc ggtctgagca ttgaaagcgt tgcacgtcgt gccggtgcaa
      421 gcaaaccgac catttatcgt tggtggacca ataaagcagc actgattgcc gaagtgtatg
      481 aaaatgaaag cgaacaggtg cgtaaatttc cggatctggg tagctttaaa gccgatctgg
      541 attttctgct gcgtaatctg tggaaagttt ggcgtgaaac catttgtggt gaagcatttc
      601 gttgtgttat tgcagaagca cagctggacc ctgcaaccct gacccagctg aaagatcagt
      661 ttatggaacg tcgtcgtgag atgccgaaaa aactggttga aaatgccatt agcaatggtg
      721 aactgccgaa agataccaat cgtgaactgc tgctggatat gatttttggt ttttgttggt
      781 atcgcctgct gaccgaacag ctgaccgttg aacaggatat tgaagaattt accttcctgc
      841 tgattaatgg tgtttgtccg ggtacacagc gttaataggt ccaggcatca aataaaacga
      901 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
      961 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatac gctatgtctt
     1021 ctactagtag cggccgctgc agtccggcaa aaaagggcaa ggtgtcacca ccctgccctt
     1081 tttctttaaa accgaaaaga ttacttcgcg ttatgcaggc ttcctcgctc actgactcgc
     1141 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaattcatg
     1201 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1261 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1321 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1381 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     1441 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1501 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1561 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1621 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1681 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1741 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1801 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1861 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1921 ttctttcctg cgttatcccc tgattctgtg gataaccgtc gattatcaaa aaggatcttc
     1981 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2041 acttggtctg acagctcgag gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
     2101 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
     2161 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa
     2221 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     2281 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
     2341 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     2401 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     2461 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     2521 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     2581 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
     2641 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     2701 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     2761 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     2821 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     2881 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc
     2941 gatcaactcg agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     3001 aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg
     3061 atgatttctg gaattcgcgg ccgcttctag ag
//
