LOCUS       62056_14070        8814 bp DNA     circular SYN 16-DEC-2019
DEFINITION  Cloning vector pLG231, complete sequence.
ACCESSION   MN443779
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8814)
  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 8814)
  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 8814)
  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..8814
                     /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"
     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"
     terminator      5553..5587
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     misc_binding    5642..5661
                     /label=LacO binding site
                     /bound_moiety="LacO"
     protein_bind    5715..5731
                     /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)."
     misc_RNA        5735..5785
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      5816..5832
                     /label=SpoVG
                     /note="SpoVG"
                     /regulatory_class="ribosome_binding_site"
     CDS             5836..6543
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     terminator      6599..6693
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      6801..6856
                     /label=Ppen
                     /note="Ppen"
                     /regulatory_class="promoter"
     CDS             6899..7978
                     /label=lacI
                     /note="lac repressor"
     misc_feature    8147..8814
                     /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 cgccgggcgt
     5581 tttttatgca gcaatggcaa gaacgttgct cgagggtctc ccgcttaacg atcgttggct
     5641 gaaatgtgag cactcacaat tcattttgca aaagttgttg actttatcta caaggtgtgg
     5701 cataatgtgt gtaattgtga gcggataaca attagctgtc accggatgtg ctttccggtc
     5761 tgatgagtcc gtgaggacga aacagcctct acaaataatt ttgtttaatc ggctgaaagg
     5821 tggtgaacta ctatgaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa
     5881 ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca
     5941 acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg
     6001 ccaacacttg tcactacttt cgcgtatggt cttcaatgct ttgcgagata cccagatcat
     6061 atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact
     6121 atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat
     6181 acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt
     6241 ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa
     6301 aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     6361 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     6421 aaccattacc tgtccacaca atctaagctt tcgaaagatc ccaacgaaaa gagggaccac
     6481 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
     6541 aaatactagt aagcttgcgg ccgcgtcgtg actgggaaaa ccctggcgac tagtcttgga
     6601 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
     6661 ggttgccgcc gggcgttttt tattggtgag aatccagggg tccccaataa ttacgattta
     6721 aattggagct tagtcgacag ctagccgcat gcaagctaat tcggtggaaa cgaggtcatc
     6781 atttccttcc gaaaaaacgg ttgcatttaa atcttacata tgtaatactt tcaaagacta
     6841 catttgtaag atttgatgtt tgtagcgccc ggaagagagt caattcaggg tggtgaatat
     6901 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttata tgaccgtttc
     6961 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     7021 gatggtggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     7081 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     7141 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     7201 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     7261 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     7321 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattattta
     7381 ctcccatgag gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     7441 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     7501 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
     7561 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     7621 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     7681 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag atagctcatg
     7741 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     7801 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgccagt
     7861 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     7921 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     7981 ataaggatcc taattggtaa cgaatcagac aattgacggc gcgcaacgca attaatgtga
     8041 gttaggatcc tgagcgccgg tcgctaccat taccagttgg tctggtgtca aaaataataa
     8101 taaccgggca ggccatgtct gcccgtattt cgcgtaagga aatccattat gtactattta
     8161 attctgcgtg acatcccatc gatcagacca gtttttaatt tgtgtgtttc catgtgtcca
     8221 gtttggaata ctcttaacct cattggaaat cgcggcataa tcactggtgg tatgattgat
     8281 gaccgcgtca acaatgacct ttatgccata ttcttcagcg gctgcacaca tttctttaaa
     8341 ttcttgttca gtacctaagt aacggttgcc aatttgatac gatgtcggct gatacagcca
     8401 gtaccagttc gacatgcttt tatctccttg attcccttcc tttacttggt taatcggaga
     8461 tgtctgaatg gctgtatatc ctgcatcatg aatatccttc atattgtgtt ttaacgtatt
     8521 gaacgaccaa ttccatgcat gaagaatggt tccgcttttg atcgacggtg ctgtaagctc
     8581 attcgatttg ttcgccgttt cagcactcgc agccgccggt cctgccagaa ccaaatgaaa
     8641 cagcaataaa aatccagcga ataacggcag taaagaggtt ttgaatcgtt ttgcaaacat
     8701 tcttgacact ccttatttga ttttttgaag acttacttcg gagtcaaaaa tccctcttac
     8761 ttcattcttc cgcttcctcc tttcaaaccg atgtgaagac tggagaattt tgtt
//
