LOCUS       62056_14075        8388 bp DNA     circular SYN 16-DEC-2019
DEFINITION  Cloning vector pLG320, complete sequence.
ACCESSION   MN443780
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8388)
  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 8388)
  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 8388)
  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..8388
                     /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
                     /direction=RIGHT
                     /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"
     misc_binding    5593..5604
                     /label=VanO binding site
                     /bound_moiety="VanO"
     regulatory      5621..5649
                     /label=pSpank
                     /note="pSpank"
                     /regulatory_class="promoter"
     misc_binding    5647..5658
                     /label=VanO binding site
                     /bound_moiety="VanO"
     misc_RNA        5660..5710
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      5741..5752
                     /label=SpoVG
                     /note="SpoVG"
                     /regulatory_class="ribosome_binding_site"
     CDS             5761..6468
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     terminator      6524..6618
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      6726..6796
                     /label=Ppen
                     /note="Ppen"
                     /regulatory_class="promoter"
     regulatory      6804..6815
                     /label=SpoVG
                     /note="SpoVG"
                     /regulatory_class="ribosome_binding_site"
     CDS             6821..7555
                     /codon_start=1
                     /transl_table=11
                     /product="VanR-IVS"
                     /label=VanR-IVS
                     /protein_id="QGV56734.1"
                     /translation="MDMPRIKPGQRVMMALRKMIASGEIKSGERIAEIPTAAALGVSRM
                     PVRIALRSLEQEGLVVRLGARGYAARGVSSDQIRDAIEVRGVLEGFAARRLAERGMTAE
                     THARFVVLIAEGEALFAAGRLNGEDLDRYAAYNQAFHDTLVSAAGNGAVESALARNGFE
                     PFAAAGALALDLMDLSAEYEHLLAAHRQHQAVLDAVSCGDAEGAERIMRDHALAAIRNA
                     KVFEAAASAGAPLGAAWSIRAD"
     misc_feature    7721..8388
                     /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 cgcggcaaga
     5581 acgttgctcg agattggatc caatcatttt gcaaaagttg ttgactttat ctacaaggtg
     5641 tggcatattg gatccaatag ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag
     5701 gacgaaacag cctctacaaa taattttgtt taatcggctg aaaggtggtg aactactatg
     5761 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     5821 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
     5881 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     5941 actttcgcgt atggtcttca atgctttgcg agatacccag atcatatgaa acagcatgac
     6001 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     6061 gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga
     6121 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
     6181 tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa
     6241 gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     6301 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     6361 acacaatcta agctttcgaa agatcccaac gaaaagaggg accacatggt ccttcttgag
     6421 tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata ctagtaagct
     6481 tgcggccgcg tcgtgactgg gaaaaccctg gcgactagtc ttggactcct gttgatagat
     6541 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     6601 ttttttattg gtgagaatcc aggggtcccc aataattacg atttaaattg gagcttagtc
     6661 gacagctagc cgcatgcaag ctaattcggt ggaaacgagg tcatcatttc cttccgaaaa
     6721 aacggttgca tttaaatctt acatatgtaa tactttcaaa gactacattt gtaagatttg
     6781 atgtttgtag cgcccgtcgg ctgaaaggtg gtgaactact atggacatgc ctcgtattaa
     6841 accgggtcag cgtgttatga tggcactgcg taaaatgatt gcaagcggtg aaatcaaaag
     6901 tggtgaacgt attgcagaaa ttccgaccgc agcagcactg ggtgttagcc gtatgccggt
     6961 tcgtatcgca ctgcgttcac tggaacaaga aggtctggtt gttcgtctgg gtgcacgtgg
     7021 ttatgcagcc cgtggtgtta gcagcgatca gattcgtgat gcaattgaag ttcgtggtgt
     7081 tctggaaggt tttgcagcac gtcgtctggc agaacgtggt atgaccgcag aaacccatgc
     7141 acgttttgtt gtactgattg cagaaggtga agcactgttt gcagccggtc gcctgaatgg
     7201 tgaagatctg gatcgttatg ccgcatataa tcaggcattt catgataccc tggttagcgc
     7261 agcaggtaat ggtgcagttg aaagcgcact ggcacgtaat ggttttgaac cgtttgcagc
     7321 agccggtgca ctggccctgg atctgatgga cctgtctgcc gaatatgaac atctgctggc
     7381 agcacatcgt cagcatcagg cagttctgga tgcagttagc tgtggtgatg ccgaaggtgc
     7441 agaacgtatt atgcgtgatc atgcactggc agcaattcgt aatgcaaaag tttttgaagc
     7501 agcagcaagc gcaggcgcac cgctgggtgc agcatggtca attcgtgcag attgataagg
     7561 atcctaattg gtaacgaatc agacaattga cggcgcgcaa cgcaattaat gtgagttagg
     7621 atcctgagcg ccggtcgcta ccattaccag ttggtctggt gtcaaaaata ataataaccg
     7681 ggcaggccat gtctgcccgt atttcgcgta aggaaatcca ttatgtacta tttaattctg
     7741 cgtgacatcc catcgatcag accagttttt aatttgtgtg tttccatgtg tccagtttgg
     7801 aatactctta acctcattgg aaatcgcggc ataatcactg gtggtatgat tgatgaccgc
     7861 gtcaacaatg acctttatgc catattcttc agcggctgca cacatttctt taaattcttg
     7921 ttcagtacct aagtaacggt tgccaatttg atacgatgtc ggctgataca gccagtacca
     7981 gttcgacatg cttttatctc cttgattccc ttcctttact tggttaatcg gagatgtctg
     8041 aatggctgta tatcctgcat catgaatatc cttcatattg tgttttaacg tattgaacga
     8101 ccaattccat gcatgaagaa tggttccgct tttgatcgac ggtgctgtaa gctcattcga
     8161 tttgttcgcc gtttcagcac tcgcagccgc cggtcctgcc agaaccaaat gaaacagcaa
     8221 taaaaatcca gcgaataacg gcagtaaaga ggttttgaat cgttttgcaa acattcttga
     8281 cactccttat ttgatttttt gaagacttac ttcggagtca aaaatccctc ttacttcatt
     8341 cttccgcttc ctcctttcaa accgatgtga agactggaga attttgtt
//
