LOCUS       40924_43313        5346 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTHSSe_57, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5346)
  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 5346)
  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 5346)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5346)
  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..5346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..199)
                     /label=Tn7 right end
                     /note="Tn7 right end"
     terminator      complement(1294..1337)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1340..1361
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      1374..1421
                     /note="PJ23105; derived from BBa_J23105"
                     /regulatory_class="promoter"
     misc_RNA        1423..1473
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      1497..1515
                     /note="B0032; derived from BBa_B0032"
                     /regulatory_class="ribosome_binding_site"
     CDS             1516..2229
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     terminator      2249..2320
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2336..2363
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             2633..3421
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     mobile_element  complement(3517..3682)
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        3751..3842
                     /label=AmpR promoter
     CDS             3843..4499
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      4506..4545
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            4693..4801
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      4812..5200
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     terminator      complement(5231..5317)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 tgtgggcgga caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt
       61 cttaaactag acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat
      121 actggacttt tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc
      181 gtattaaaga ggggcgtggt tgtgtctcag gccgcctagg ccagacttaa gccagttcat
      241 tgcttcaacc gcagctacca ccgtcacgct cacagattcc caaatcagct tagccatgta
      301 tccagcagcc tgattcggct gagtgaacat cagacccttg ccggaatcaa tagctggctg
      361 aatggtatct tccagcactt gttgacggaa gccgaactct ttggacccgt aagccagcgt
      421 catgactgaa ctcttagtca cactgcgagt aacaccgtaa gccagccatt gaccagccag
      481 tgccttagtg cccagcttga ctttctcaga gatttcacca gtgttctcat cggtcacggt
      541 aactacttcg ttatcggtcc cattgattgc gtctgcttgt agaatctcgt tgactttctt
      601 agcaacaatc ccgtagatgt cctgaacggt ttcactagga agcaagttaa ccgcgcgacc
      661 acctacctca tctcggagca tcgcggagaa gtgctggatg ccagagcaag acccgtcaaa
      721 ttacgccagc ggaagggagc agttatagct caggccgtgg tgctgtaccc cagcgtactc
      781 aaagcagaac gcaaggaagc agaacggaga atcttgctca gcccaccaag tgttctccag
      841 tggagactta gcgcaagcca tgatgttctc gtggttttcc tcaatgaact tgatgcgctc
      901 agggaacgga accttgtcga cacccgcaca gtttgcaccg tggattttca gccagtagta
      961 accttcctta ccgattggtt tacctttcgc cagcgtaagc agtcctttgg tcatatcgtt
     1021 accttgcggg ttgaacattg acacagcgta aacacgaccg cgccagtcca tgttgtaagg
     1081 gaaccagatg gccttatggt tagcaaactt attggcttgc tcaagcatga actcaaggct
     1141 gatacggcga gacttgcgag ccttgtcctt gcggtacaca gcagcggcag cacgtttcca
     1201 cgcggtgaga gcctcaggat ttgccacctg acgtctaaga aaccattatt atcatgacat
     1261 taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt
     1321 tcgctaagga tgatttctgg aattcgcggc cgcttctaga gggaggtgtc ttcggcgcgc
     1381 ctttacggct agctcagtcc taggtactat gctagcaagg tagctgtcac cggatgtgct
     1441 ttccggtctg atgagtccgt gaggacgaaa cagcctctac aaataatttt gtttaatcac
     1501 acaggaaagt actagatgag caaaggagaa gaacttttca ctggagttgt cccaattctt
     1561 gttgaattag atggtgatgt taatgggcac aaattttctg tccgtggaga gggtgaaggt
     1621 gatgctacaa acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt
     1681 ccgtggccaa cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg
     1741 gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
     1801 cgcactatat ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt caagtttgaa
     1861 ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac
     1921 attcttggac acaaactcga gtacaacttt aactcacaca atgtatacat cacggcagac
     1981 aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacgttga agatggttcc
     2041 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     2101 ccagacaacc attacctgtc gacacaatct gtcctttcga aagatcccaa cgaaaagcgt
     2161 gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag
     2221 ctctacaaat aatactagag ccaggcatca aataaaacga aaggctcagt cgaaagactg
     2281 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     2341 accttcgggt gggcctttct gcgtttatag aagacatcgc ttactagtag cggccgccaa
     2401 agcccgccga aaggcgggct tttctgtatt taaatgaacc ttgaccgaac gcagcggtgg
     2461 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     2521 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     2581 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatgaggga
     2641 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     2701 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     2761 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     2821 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     2881 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     2941 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     3001 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     3061 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     3121 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     3181 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     3241 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     3301 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     3361 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     3421 ataagacaat tgtctaatta attgcggacc ctagaggtcc ccttttttat tttaaaaatt
     3481 ttttcacaaa acggtttaca agcataaaat ctctgaaacc agataagtga aatctagttc
     3541 caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat
     3601 ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc
     3661 ccaactattt tgtccgccca cagcggggca tttttcttcg gcgatcgcta gtagcccgcc
     3721 taatgagcgg gctttttttt aattccccta tttgtttatt tttctaaata cattcaaata
     3781 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3841 gtatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     3901 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     3961 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     4021 ttcacattct tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg
     4081 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     4141 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     4201 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     4261 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     4321 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     4381 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     4441 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     4501 aaaccgatac aattaaaggc tccttttgga gccttttttt ttggacacgc gtttgtcctt
     4561 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     4621 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     4681 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     4741 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     4801 gtaggccggc cgatctgaag atcagcagtt caacctgttg atagtacgta ctaagctctc
     4861 atgtttcacg tactaagctc tcatgtttaa cgtactaagc tctcatgttt aacgaactaa
     4921 accctcatgg ctaacgtact aagctctcat ggctaacgta ctaagctctc atgtttcacg
     4981 tactaagctc tcatgtttga acaataaaat taatataaat cagcaactta aatagcctct
     5041 aaggttttaa gttttataag aaaaaaaaga atatataagg cttttaaagc ctttaaggtt
     5101 taacggttgt ggacaacaag ccagggatgt aacgcactga gaagccctta gagcctctca
     5161 aagcaatttt gagtgacaca ggaacactta acggctgaca tggggcgcgc ccagctgtct
     5221 agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     5281 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cctttaatta accctgcgta
     5341 agcggg
//
