LOCUS       62056_14050        8591 bp DNA     circular SYN 16-DEC-2019
DEFINITION  Cloning vector pLG163, complete sequence.
ACCESSION   MN443775
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8591)
  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 8591)
  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 8591)
  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..8591
                     /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..4139
                     /label=similar to AmyE
                     /note="similar to AmyE"
     CDS             complement(4266..5018)
                     /codon_start=1
                     /transl_table=11
                     /product="SpecR"
                     /label=SpecR
                     /protein_id="QGV56725.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     terminator      5347..5433
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5452..5479
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      5559..5593
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     regulatory      5660..5688
                     /label=pSpank
                     /note="pSpank"
                     /regulatory_class="promoter"
     protein_bind    5697..5713
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     regulatory      5729..5745
                     /label=SpoVG
                     /note="SpoVG"
                     /regulatory_class="ribosome_binding_site"
     CDS             5749..6456
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     CDS             6715..7794
                     /label=lacI
                     /note="lac repressor"
     misc_feature    7924..8591
                     /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 gcgtgggtct 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 tatcaactct 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 agtgatcgaa 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 tgccaaattg gatctccttt
     3541 tccgattgtg aattgatctc catccttaaa cgcctgtcgt ctggtccatt attgatttga
     3601 taaacggctt ttgttgtatt cgcatctgca cgcaaggtaa tcgtcagttg atcattgaaa
     3661 gaatgtgtta cacctgtttt gtaattctca aggaaaacat gaggcgcttt tgcaatatca
     3721 tcaggataaa gcacagctac agacctggca ttgatcgtgc ctgtcagttt accatcgttc
     3781 acttgaaatg aacccgctcc agctttattg tcatacctgc catcaggcaa ttttgttgcc
     3841 gtattgatag agacagagga tgaacctgca tttgccagca caacgccatg tgagccgcgc
     3901 tgattcataa atatctggtt gtttccattc gggttcgaga gttcctcagg ctgtccagcc
     3961 atcacattgt gaaatctatt gaccgcagtg atagcctgat cttcaaataa agcactcccg
     4021 cgatcgccta tttggctttt ccccgggaac ctcacaccat ttccgcctcc ctcaggtctg
     4081 gaaaagaaaa gaggcgtact gcctgaacga gaagctatca ccgcccagcc taaacggatg
     4141 atccccctat gcaagggttt attgttttct aaaatctgat taccaattag aatgaatatt
     4201 tcccaaatat taaataataa aacaaaaaaa ttgaaaaaag tgtttccacc attttttcaa
     4261 tttttttata atttttttaa tctgttattt aaatagttta tagttaaatt tacattttca
     4321 ttagtccatt caatattctc tccaagataa ctacgaactg ctaacaaaat tctctcccta
     4381 tgttctaatg gagaagattc agccactgca tttcccgcaa tatcttttgg tatgatttta
     4441 cccgtgtcca tagttaaaat catacggcat aaagttaata tagagttggt ttcatcatcc
     4501 tgataattat ctattaattc ctctgacgaa tccataatgg ctcttctcac atcagaaaat
     4561 ggaatatcag gtagtaattc ctctaagtca taatttccgt atattctttt attttttcgt
     4621 tttgcttggt aaagcattat ggttaaatct gaatttaatt ccttctgagg aatgtatcct
     4681 tgttcataaa gctcttgtaa ccattctcca taaataaatt cttgtttggg aggatgattc
     4741 cacggtacca tttcttgctg aataataatt gttaattcaa tatatcgtaa gttgctttta
     4801 tctcctattt tttttgaaat aggtctaatt ttttgtataa gtatttcttt actttgatct
     4861 gtcaatggtt cagatacgac gactaaaaag tcaagatcac tatttggttt tagtccactc
     4921 tcaactcctg atccaaacat gtaagtacca ataaggttat tttttaaatg tttccgaagt
     4981 atttttttca ctttattaat ttgttcgtat gtattcaaat atatcctcct cactattttg
     5041 attagtacct attttatatc catagttgtt aattaaataa acttaattta gtttatttat
     5101 agatttcatt ggcttctaaa ttttttatct agataataat tattttagtt aattttattc
     5161 tagattatat atgatatgat ctttcatttc cataaaacta aagtaagtgt aaacctattc
     5221 attgttttaa aaatatctct tgccagtcac gttacgttat tagttatagt tattataaca
     5281 tgtattcacg aacgaaaatc gccattcgcc agggctgcag gaattcgact ctctagcttg
     5341 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     5401 ttgtcggtga acgctctcct gagtaggaca aatccgccgc tctagctaag cagaaggcca
     5461 tcctgacgga tggccttttt gcgtttctac aaactcttgt taactctaga gctgcctgcc
     5521 gcgtttcggt gatgaagatc ttcccgatga ttaattaatt cagaacgctc ggttgccgcc
     5581 gggcgttttt tatgcagcaa tggcaagaac gttgctcgag ggtaaatgtg agcactcaca
     5641 attcattttg caaaagttgt tgactttatc tacaaggtgt ggcataatgt gtgtaattgt
     5701 gagcggataa caattaagct ttcggctgaa aggtggtgaa ctactatgaa aggagaagaa
     5761 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
     5821 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
     5881 atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac tttcgcgtat
     5941 ggtcttcaat gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt
     6001 gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac
     6061 aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa
     6121 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac
     6181 tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa
     6241 attagacaca acattgaaga tggaagcgtt caactagcag accattatca acaaaatact
     6301 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctaag
     6361 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
     6421 gctgggatta cacatggcat ggatgaacta tacaaataag ctagccgcat gcaagctaat
     6481 tcggtggaaa cgaggtcatc atttccttcc gaaaaaacgg ttgcatttaa atcttacata
     6541 tgtaatactt tcaaagacta catttgtaag atttgatgtt tgagtcggct gaaagatcgt
     6601 acgtaccaat tattgtttcg tgattgttca agccataaca ctgtagggat agtggaaaga
     6661 gtgcttcatc tggttacgat caatcaaata ttcaaacgga gggagacgat tttgatgaaa
     6721 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     6781 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     6841 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     6901 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     6961 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     7021 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     7081 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     7141 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     7201 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     7261 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     7321 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     7381 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     7441 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     7501 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     7561 atcccgccgt caaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     7621 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     7681 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     7741 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     7801 caacgcaatt aatgtgagtt aggatcctga gcgccggtcg ctaccattac cagttggtct
     7861 ggtgtcaaaa ataataataa ccgggcaggc catgtctgcc cgtatttcgc gtaaggaaat
     7921 ccattatgta ctatttaatt ctgcgtgaca tcccatcgat cagaccagtt tttaatttgt
     7981 gtgtttccat gtgtccagtt tggaatactc ttaacctcat tggaaatcgc ggcataatca
     8041 ctggtggtat gattgatgac cgcgtcaaca atgaccttta tgccatattc ttcagcggct
     8101 gcacacattt ctttaaattc ttgttcagta cctaagtaac ggttgccaat ttgatacgat
     8161 gtcggctgat acagccagta ccagttcgac atgcttttat ctccttgatt cccttccttt
     8221 acttggttaa tcggagatgt ctgaatggct gtatatcctg catcatgaat atccttcata
     8281 ttgtgtttta acgtattgaa cgaccaattc catgcatgaa gaatggttcc gcttttgatc
     8341 gacggtgctg taagctcatt cgatttgttc gccgtttcag cactcgcagc cgccggtcct
     8401 gccagaacca aatgaaacag caataaaaat ccagcgaata acggcagtaa agaggttttg
     8461 aatcgttttg caaacattct tgacactcct tatttgattt tttgaagact tacttcggag
     8521 tcaaaaatcc ctcttacttc attcttccgc ttcctccttt caaaccgatg tgaagactgg
     8581 agaattttgt t
//
