LOCUS       62056_14085        8233 bp DNA     circular SYN 16-DEC-2019
DEFINITION  Cloning vector pLG371, complete sequence.
ACCESSION   MN443782
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8233)
  AUTHORS   Gonzalez LM, Mukhitov N, Voigt CA.
  TITLE     Resilient living materials built by printing bacterial spores
  JOURNAL   Nat. Chem. Biol. (2019) In press
  PUBMED    31792444
REFERENCE   2  (bases 1 to 8233)
  AUTHORS   Mukhitov N, Gonzalez LM, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-SEP-2019) Biological Engineering, MIT, NE47-140, 500 
            Tech Square, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 8233)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol. (2019) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-SEP-2019) Biological Engineering, MIT, NE47-140, 500 Tech 
            Square, Cambridge, MA 02139, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        106..210
                     /label=AmpR promoter
     CDS             211..1068
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1242..1830
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2963..4138
                     /label=similar to AmyE
                     /note="similar to AmyE"
     gene            complement(4263..5045)
                     /gene="specR"
                     /label=specR
     terminator      5341..5427
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5446..5473
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     protein_bind    5574..5601
                     /label=CuO
                     /note="CymR-binding P2 operator sequence from the p-cmt
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     regulatory      5606..5634
                     /label=pSpank
                     /note="pSpank"
                     /regulatory_class="promoter"
     misc_binding    5635..5666
                     /label=CymO binding site
                     /bound_moiety="CymO"
     misc_RNA        5668..5718
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      5749..5760
                     /label=SpoVG
                     /note="SpoVG"
                     /regulatory_class="ribosome_binding_site"
     CDS             5769..6476
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     terminator      6532..6626
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      6734..6804
                     /label=Ppen
                     /note="Ppen"
                     /regulatory_class="promoter"
     regulatory      6812..6828
                     /label=SpoVG
                     /note="SpoVG"
                     /regulatory_class="ribosome_binding_site"
     CDS             6829..7437
                     /label=CymR
                     /note="cumate repressor (Mullick et al., 2006)"
     misc_feature    7599..8233
                     /label=similar to AmyE
                     /note="similar to AmyE"
ORIGIN
        1 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
       61 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      121 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      181 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      241 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      301 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      361 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
      421 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
      481 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
      541 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
      601 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
      661 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
      721 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
      781 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
      841 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
      901 gataaatctg gagccggtga gcgtgggtcg cgcggtatca ttgcagcact ggggccagat
      961 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1021 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1081 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1141 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1381 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     1741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     1801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcccg acctcgagct
     1861 ggatacttcc cgtccgccag ggggacatgc cggcgatgct gaaggtcgcg cgcattcccg
     1921 atgaagaggc cggttaccgc ctgtttgagg atatagtaat ctttctaaat agctttggat
     1981 tggaggagta tggccactaa tactaagttc agctaataaa aaaatttgct aaagaactcc
     2041 agctggattt cactgatgag aatatcgtcg gagataaata taataattcc acggactata
     2101 gactatacta gtatactccg tctactgtac gatacacttc cgctcaggtc cttgtccttt
     2161 aacgaggatt gttacgtacg ctaatggcgt caaaacaaag actctagacc taggccttaa
     2221 gatctgatca tatgcatccg cgggcccggg ttaacgcgta atccatggat caagagacag
     2281 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggtgccct
     2341 gaatgaactg cagaaagagc tggtagttgg cgcactgttc gaagaactgc cgatgtccag
     2401 taagattctt actatgctgg ttgaaccgga tgctggtaaa gctacttggg ttgctgcttc
     2461 tacttatggt accgatacaa ctactggtga ggaagttaaa ggagctctta aagaaatcca
     2521 cttcagtagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     2581 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     2641 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     2701 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     2761 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     2821 gcgactaaga aaatgccgtc aaatccgctc gccatgactt cactaacgat gcctttgaaa
     2881 atcttcaagt tcttttctac taattcaagg cgtgtctcac caggtttttg gtttgctccg
     2941 gcgcaaatgc agacaatatc agcatccttg cagggtatgt ttctctttga tgtctttttg
     3001 tttgtgaagt atttcacatt tatattgtgc aacacttcac aaacttttgc aagagaaaag
     3061 ttttgtctga tttatgaaca aaaaagaaac catcattgat ggtttctttc ggtaagtccc
     3121 gtctagcctt gccctcaatg gggaagagaa ccgcttaagc ccgagtcatt atataaacca
     3181 tttagcacgt aatcaaagcc aggctgattc tgaccgggca cttgggcgct gccattatta
     3241 aaaatcactt ttgcgttggt tgtatccgtg tccgcaggca gcgtcagcgt gtaaattccg
     3301 tctgcatttt tagtcattgg ttttccaggc caagatccgg tcaattcaat tactcggctc
     3361 ccatcatgtt tatagatata agcatttacc tggctccaat gattcggatt ttgatagccg
     3421 atggttttgg ccgacgctgg atctctttta acaaaactgt atttctcggt cctcgttaca
     3481 ccatcactgt tcgttccttt taacatgatg gtgtatgttt tgccaaatgg atctcctttt
     3541 ccgattgtga attgatctcc atccttaaac gcctgtcgtc tggtccatta ttgatttgat
     3601 aaacggcttt tgttgtattc gcatctgcac gcaaggtaat cgtcagttga tcattgaaag
     3661 aatgtgttac acctgttttg taattctcaa ggaaaacatg aggcgctttt gcaatatcat
     3721 caggataaag cacagctaca gacctggcat tgatcgtgcc tgtcagttta ccatcgttca
     3781 cttgaaatga acccgctcca gctttattgt catacctgcc atcaggcaat tttgttgccg
     3841 tattgataga gacagaggat gaacctgcat ttgccagcac aacgccatgt gagccgcgct
     3901 gattcataaa tatctggttg tttccattcg ggttcgagag ttcctcaggc tgtccagcca
     3961 tcacattgtg aaatctattg accgcagtga tagcctgatc ttcaaataaa gcactcccgc
     4021 gatcgcctat ttggcttttc cccgggaacc tcacaccatt tccgcctccc tcaggtctgg
     4081 aaaagaaaag aggcgtactg cctgaacgag aagctatcac cgcccagcct aaacggattc
     4141 cccctatgca agggtttatt gttttctaaa atctgattac caattagaat gaatatttcc
     4201 caaatattaa ataataaaac aaaaaaattg aaaaaagtgt ttccaccatt ttttcaattt
     4261 ttttataatt tttttaatct gttatttaaa tagtttatag ttaaatttac attttcatta
     4321 gtccattcaa tattctctcc aagataacta cgaactgcta acaaaattct ctccctatgt
     4381 tctaatggag aagattcagc cactgcattt cccgcaatat cttttggtat gattttaccc
     4441 gtgtccatag ttaaaatcat acggcataaa gttaatatag agttggtttc atcatcctga
     4501 taattatcta ttaattcctc tgacgaatcc ataatggctc ttctcacatc agaaaatgga
     4561 atatcaggta gtaattcctc taagtcataa tttccgtata ttcttttatt ttttcgtttt
     4621 gcttggtaaa gcattatggt taaatctgaa tttaattcct tctgaggaat gtatccttgt
     4681 tcataaagct cttgtaacca ttctccataa ataaattctt gtttgggagg atgattccac
     4741 ggtaccattt cttgctgaat aataattgtt aattcaatat atcgtaagtt gcttttatct
     4801 cctatttttt ttgaaatagg tctaattttt tgtataagta tttctttact ttgatctgtc
     4861 aatggttcag atacgacgac taaaaagtca agatcactat ttggttttag tccactctca
     4921 actcctgatc caaacatgta agtaccaata aggttatttt ttaaatgttt ccgaagtatt
     4981 tttttcactt tattaatttg ttcgtatgta ttcaaatata tcctcctcac tattttgatt
     5041 agtacctatt ttatatccat agttgttaat taaataaact taatttagtt tatttataga
     5101 tttcattggc ttctaaattt tttatctaga taataattat tttagttaat tttattctag
     5161 attatatatg atatgatctt tcatttccat aaaactaaag taagtgtaaa cctattcatt
     5221 gttttaaaaa tatctcttgc cagtcacgtt acgttattag ttatagttat tataacatgt
     5281 attcacgaac gaaaatcgcc attcgccagc tgcaggaatt cgacctctag cttgaggcat
     5341 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     5401 gtgaacgctc tcctgagtag gacaaatccg ccgctctagc taagcagaag gccatcctga
     5461 cggatggcct ttttgcgttt ctacaaactc ttgttaactc tagagctgcc tgccgcgttt
     5521 cggtgatgaa gatcttcccg atgattaatt aattcagaac gctcggttgc cgcaacaaac
     5581 agacaatctg gtctgtttgt attatttgac tttatctaca aggtgtggca taataacaaa
     5641 cagacaatct ggtctgtttg tattatagct gtcaccggat gtgctttccg gtctgatgag
     5701 tccgtgagga cgaaacagcc tctacaaata attttgttta atcggctgaa aggtggtgaa
     5761 ctactatgaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
     5821 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
     5881 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac
     5941 ttgtcactac tttcgcgtat ggtcttcaat gctttgcgag atacccagat catatgaaac
     6001 agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt
     6061 tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg
     6121 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca
     6181 aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg
     6241 gaatcaaagt taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag
     6301 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
     6361 acctgtccac acaatctaag ctttcgaaag atcccaacga aaagagggac cacatggtcc
     6421 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatact
     6481 agtaagcttg cggccgcgtc gtgactggga aaaccctggc gactagtctt ggactcctgt
     6541 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     6601 gccgggcgtt ttttattggt gagaatccag gggtccccaa taattacgat ttaaattgga
     6661 gcttagtcga cagctagccg catgcaagct aattcggtgg aaacgaggtc atcatttcct
     6721 tccgaaaaaa cggttgcatt taaatcttac atatgtaata ctttcaaaga ctacatttgt
     6781 aagatttgat gtttgtagcg cccgtcggct gaaaggtggt gaactactat gagcccgaaa
     6841 cgtcgtaccc aggcagaacg tgcaatggaa acccagggta aactgattgc agcagcactg
     6901 ggtgttctgc gtgaaaaagg ttatgcaggt tttcgtattg cagatgttcc gggtgcagcc
     6961 ggtgttagcc gtggtgcaca gagccatcat tttccgacca aactggaact gctgctggca
     7021 acctttgaat ggctgtatga gcagattacc gaacgtagcc gtgcacgtct ggcaaaactg
     7081 aaaccggaag atgatgttat tcagcagatg ctggatgatg cagcagaatt ttttctggat
     7141 gatgatttta gcatcggcct ggatctgatt gttgcagcag atcgtgatcc ggcactgcgt
     7201 gaaggtattc agcgtaccgt tgaacgtaat cgttttgttg ttgaagatat gtggctgggt
     7261 gtgctggtga gccgtggtct gagccgtgat gatgccgaag atattctgtg gctgattttt
     7321 aacagcgttc gtggtctggt agttcgtagc ctgtggcaga aagataaaga acgttttgaa
     7381 cgtgtgcgta atagcaccct ggaaattgca cgtgaacgtt atgcaaaatt caaacgttga
     7441 taaggatcct aattggtaac gaatcagaca attgacggcg cgcaacgcaa ttaatgtgag
     7501 ttaggatcct gagcgccggt cgctaccatt accagttggt ctggtgtcaa aaataataat
     7561 aaccgggcag gccatgtctg cccgtatttc gcgtaaggcg atcagaccag tttttaattt
     7621 gtgtgtttcc atgtgtccag tttggaatac tcttaacctc attggaaatc gcggcataat
     7681 cactggtggt atgattgatg accgcgtcaa caatgacctt tatgccatat tcttcagcgg
     7741 ctgcacacat ttctttaaat tcttgttcag tacctaagta acggttgcca atttgatacg
     7801 atgtcggctg atacagccag taccagttcg acatgctttt atctccttga ttcccttcct
     7861 ttacttggtt aatcggagat gtctgaatgg ctgtatatcc tgcatcatga atatccttca
     7921 tattgtgttt taacgtattg aacgaccaat tccatgcatg aagaatggtt ccgcttttga
     7981 tcgacggtgc tgtaagctca ttcgatttgt tcgccgtttc agcactcgca gccgccggtc
     8041 ctgccagaac caaatgaaac agcaataaaa atccagcgaa taacggcagt aaagaggttt
     8101 tgaatcgttt tgcaaacatt cttgacactc cttatttgat tttttgaaga cttacttcgg
     8161 agtcaaaaat ccctcttact tcattcttcc gcttcctcct ttcaaaccga tgtgaagact
     8221 ggagaatttt gtt
//
