LOCUS       62056_18100        9993 bp DNA     circular SYN 16-DEC-2019
DEFINITION  Cloning vector pNM58, complete sequence.
ACCESSION   MN443783
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9993)
  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 9993)
  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 9993)
  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..9993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      251..795
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             1643..2350
                     /codon_start=1
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     terminator      2383..2430
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(6308..7165)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHF"
     promoter        complement(7166..7270)
                     /label=AmpR promoter
     CDS             complement(7437..8810)
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="VNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     CDS             complement(join(9013..9993,1..21))
                     /codon_start=1
                     /label=repB
                     /note="RepB replication protein"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
ORIGIN
        1 aatattaaaa gaaaccccca tctatttagt tatttgtttg gtcacttata actttaacag
       61 atggggtttt tctgtgcaac caattttaag ggttttcaat actttaaaac acatacatac
      121 caacacttca acgcaccttt cagcaaccct ttggtattat ttttttgtgt ggaaaatacg
      181 gctatgggag aagacttgct caccaaattt cgttccactg agcgtcagac cccttaataa
      241 gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa
      301 ccgccttgca gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta
      361 actggcttgg aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc
      421 atgacttcaa gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt
      481 gcatgtcttt ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac
      541 tgaacggggg gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt
      601 gtcaggcgtg gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa
      661 ggcaggaaca ggagagcgca cgagggagcc gccaggggaa acgcctggta tctttatagt
      721 cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg
      781 cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc
      841 tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa
      901 cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct
      961 gctgacgcac cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc
     1021 atcagtgcca acatagtaag ccagtataca ctccgctagc gctgaggtct gcctcgtgca
     1081 gcgagtcagt gagttatccc ctgattctgt ggataaccgt attaccgcct ttgagtggag
     1141 agcaagcccg tagtaaaata aaaatgacgt tatttctata tgtatcaaga taagaaagaa
     1201 caagttcaaa accatcaaaa aaagacacct tttcaggtgc tttttttatt ttataaactc
     1261 attccctgat ctcgacttcg ttcttttttt acctctcggt tatgagttag ttcaaattcg
     1321 ttctttttag gttctaaatc gtgtttttct tggaattgtg ctgttttatc ctttaccttg
     1381 tctacaaacc ccttaaaaac gtttttaaag gcttttaagc cgtctgtacg ttccttaagg
     1441 cctaactgta ggaaataaat acttaactgt taaatgtaat ttgtatttaa tattttaaca
     1501 taaaaaaatt tacagttaag aataaaaaac gactagttaa gaaaaattgg aaaataaatg
     1561 cttttagcat gttttaatat aactagatca cagagatgtg gtcagtcagt cttaattaac
     1621 caaggaggtg aaatgtacaa tgaaaggaga agaacttttc actggagttg tcccaattct
     1681 tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg
     1741 tgatgcaaca tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt
     1801 tccatggcca acacttgtca ctactttcgc gtatggtctt caatgctttg cgagataccc
     1861 agatcatatg aaacagcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     1921 aagaactata tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga
     1981 aggtgatacc cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa
     2041 cattcttgga cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga
     2101 caaacaaaag aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag
     2161 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     2221 accagacaac cattacctgt ccacacaatc taagctttcg aaagatccca acgaaaagag
     2281 agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga
     2341 actatacaaa taacctaggc tgctgccacc gctgagcaat aactagcata accccttggg
     2401 gcctctaaac gggtcttgag gggttttttg taaaaatatc ccactttatc caattttcgt
     2461 ttgttgaact aatgggtgct ttagttgaag aataaaagac cacattaaaa aatgtggtct
     2521 tttgtgtttt tttaaaggat ttgagcgtag cgaaaaatcc ttttctttct tatcttgata
     2581 ataagggtaa ctattgccgg atcgtcctca ggagtaggcg acatcgctaa ataatgatct
     2641 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccctttg
     2701 cgttcactta actaacttat aggggtaaca cttaaaaaag aatcaataac gatagaaacc
     2761 gctcctaaag caggtgcatt ttttcctaac gaagaaggca atagttcaca tttattgtct
     2821 aaatgagaat ggactctaga agaaacttcg tttttaatcg tatttaaaac aatgggatga
     2881 gattcaatta tatgatttct caagataaca gcttctatat caaatgtatt aaggatattg
     2941 gttaatccaa ttccgatata aaagccaaag ttttgaagtg catttaacat ttctacatca
     3001 tttttatttg cgcgttccac aatctctttt cgagaaatat tcttttcttc tttagagagc
     3061 gaagccagta acgctttttc agaagcatat aattcccaac agcctcgatt tccacagctg
     3121 catttgggtc cattaaaatc tatcgtcata tgacccattt ccccagaaaa accctgaaca
     3181 cctttataca attcgttgtt aataacaagt ccagttccaa ttccgatatt aatactgatg
     3241 taaacgatgt tttcatagtt ttttgtcata ccaaatactt tttcaccgta tgctcctgca
     3301 ttagcttcat tttcaacaaa aaccggaaca ttaaactcac tctcaattaa aaactgcaaa
     3361 tctttgatat tccaatttaa gttaggcatg aaaataattt gctgatgacg atctacaagg
     3421 cctggaacac aaattcctat tccgactaga ccataagggg actcaggcat atgggttaca
     3481 aaaccatgaa taagtgcaaa taaaatctct tttacttcac tagcggaaga actagacaag
     3541 tcagaagtct tctcgagaat aatatttcct tctaagtcgg ttagaattcc gttaagatag
     3601 tcgactccta tatcaatacc aatcgagtag cctgcattct tattaaaaac aagcattaca
     3661 ggtcttctgc cgcctctaga ttgccctgcc ccaatttcaa aaataaaatc tttttcaagc
     3721 agtgtattta cttgagagga gacagtagac ttgtttaatc ctgtaatctc agagagagtt
     3781 gccctggaga caggggagtt cttcaaaatt tcatctaata ttaatttttg attcattttt
     3841 tttactaaag cttgatctgc aatttgaata ataaccactc ctttgtttat ccaccgaact
     3901 aagttggtgt tttttgaagc ttgaattaga tatttaaaag tatcatatct aatattataa
     3961 ctaaattttc taaaaaaaac attgaaataa acatttattt tgtatatgac gagataaagt
     4021 tagtttattg gataaacaaa ctaactcaat taagatagtt gatggataaa cttgttcact
     4081 taaatcaaag ggggaaatga caaatggtcc aaactagtat ggaattgtta aacagttaca
     4141 actttgtttt gttcgtatta actcaaatga tattaatgtt tacaatacca gctataatta
     4201 gtggtattaa gtacagtaaa cttgattatt ttttcatcat agtaatttcg acattatcgt
     4261 tatttctatt taaaatgttt gatagcgcgt ccttaatcat attaacttca tttattatta
     4321 taatgtattt tgtcaaaatc aaatggtatt ctattttgtt gattatgact tcgcagatta
     4381 ttctatactg tgctaactac atgtatatag ttatatatgc atatatcacc aaaatttctg
     4441 atagtatatt tgtaatattc cctagctttt ttgtagttta tgtgactatt agtatactat
     4501 tctcatatat aataaataga gttctcaaaa aaattagcac accatatcta atactaaaca
     4561 aaggattttt aatagttatt tcgactatct tactgcttac tttttcatta tttttctttt
     4621 attcacaaat aaactcggat gaagctaaag taataaggca gtattctttt atttttattg
     4681 gtatcactat atttttaagt atattaacat ttgttatttc tcaatttctc cttaaagaga
     4741 tgaaatataa acgtaatcaa gaagaaattg aaacctatta tgaatataca ttgaagattg
     4801 aagctatcaa caacgaaatg cgcaagttcc gtcatgatta tgtcaatatc ttaacgacac
     4861 tttcagaata cattcgagaa gatgacatgc ctggcctacg tgattatttc aataaaaata
     4921 ttgtacctat gaaagacaat ttacaaatga atgctataaa attaaatggt atcgagaatc
     4981 ttaaagtacg tgaaattaaa ggcttaatta ctgcgaaaat tttacgtgca caagaaatga
     5041 atattccgat tagtatcgaa atacccgatg aagtaagtag cattaacttg aatatgatcg
     5101 atttaagtcg cagtattggt attattcttg ataatgcaat tgaggcatca actgaaattg
     5161 atgaccctat cattcgcgtt gcatttattg aaagtgaaaa ttcagtaacg tttattgtta
     5221 tgaataaatg cgctgatgat ataccacgca ttcatgaatt gttccaagaa agtttttcta
     5281 ctaaaggtga aggtcgtggt ttaggtctat caactttaaa agaaattgct gataatgcag
     5341 acaatgtctt attagataca attatcgaaa atggtttctt tattcaaaaa gttgaaatta
     5401 ttaacaacta gccataagga tgtgaatgta tgaaaatttt catttgcgaa gacgatccaa
     5461 aacaaagaga aaacatggtt accattatta aaaattatat aatgatagaa gaaaagccta
     5521 tggaaattgc cctcgcaact gataatcctt atgaggtgct tgagcaagct aaaaatatga
     5581 atgacatagg ctgttacttt ttagatattc aactttcaac cgatattaat ggtatcaaat
     5641 taggcagtga aattcgtaag catgacccag ttggtaacat tattttcgtt acgagtcaca
     5701 gtgaacttac ctatttaaca tttgtctaca aagttgcagc gatggatttt atttttaaag
     5761 atgatccagc tgaattaaga actcgaatta tagactgttt agaaactgca catacacgct
     5821 tacaattgtt gtctaaagat aatagcgttg aaacgattga attaaaacgt ggcagtaatt
     5881 cagtgtatgt tcaatatgat gatattatgt tttttgaatc atcaacaaaa tctcacagac
     5941 tcattgccca tttagataac cgtcaaattg aattttatgg taatttaaaa gaactgagtc
     6001 aattagatga tcgtttcttt agatgtcata atagctttgt cgtcaatcgc cataatattg
     6061 aatctataga ttcgaaagag cgaattgtct attttaaaaa taaagaacac tgctatgcat
     6121 cggtgagaaa cgttaaaaaa atataataag ataataaagt cagttaacgg cgtattcaat
     6181 tgtggatcct ctagagtcat ttatattcaa ctgttaacta aaactgtctg caaattaatc
     6241 aatgtctcta gtaaaattcc gattttatac ccactcctat tttcatcaat aggacatgat
     6301 ataataaaaa atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6361 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6421 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6481 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6541 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6601 gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6661 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6721 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6781 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6841 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6901 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
     6961 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7021 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7081 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7141 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7201 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7261 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7321 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattcctgt
     7381 tataaaaaaa ggatcaattt tgaactctct cccaaagttg atcccttaac gatttagaaa
     7441 tccctttgag aatgtttata tacattcaag gtaaccagcc aactaatgac aatgattcct
     7501 gaaaaaagta ataacaaatt actatacaga taagttgact gatcaacttc cataggtaac
     7561 aacctttgat caagtaaggg tatggataat aaaccaccta caattgcaat acctgttccc
     7621 tctgataaaa agctggtaaa gttaagcaaa ctcattccag caccagcttc ctgctgtttc
     7681 aagctacttg aaacaattgt tgatataact gttttggtga acgaaagccc acctaaaaca
     7741 aatacgatta taattgtcat gaaccatgat gttgtttcta aaagaaagga agcagttaaa
     7801 aagctaacag aaagaaatgt aactccgatg tttaacacgt ataaaggacc tcttctatca
     7861 acaagtatcc caccaatgta gccgaaaata atgacactca ttgttccagg gaaaataatt
     7921 acacttccga tttcggcagt acttagctgg tgaacatctt tcatcatata aggaaccata
     7981 gagacaaacc ctgctactgt tccaaatata attcccccac aaagaactcc aatcataaaa
     8041 ggtatatttt tccctaatcc gggatcaaca aaaggatctg ttactttcct gatatgtttt
     8101 acaaatatca ggaatgacag cacgctaacg ataagaaaag aaatgctata tgatgttgta
     8161 aacaacataa aaaatacaat gcctacagac attagtataa ttcctttgat atcaaaatga
     8221 ccttttatcc ttacttcttt ctttaataat ttcataagaa acggaacagt gataattgtt
     8281 atcataggaa tgagtagaag ataggaccaa tgaatataat gggctatcat tccaccaatc
     8341 gctggaccga ctccttctcc catggctact atcgatccaa taagaccaaa tgctttaccc
     8401 ctattttcct ttggaatata gcgcgcaact acaaccatta cgagtgctgg aaatgcagct
     8461 gcaccagccc cttgaataaa acgagccata ataagtaagg aaaagaaaga atggccaaca
     8521 aacccaatta ccgacccgaa acaatttatt ataattccaa ataggagtaa ccttttgatg
     8581 cctaattgat cagatagctt tccatataca gctgttccaa tggaaaaggt taacataaag
     8641 gctgtgttca cccagtttgt actcgcaggt ggtttattaa aatcatttgc aatatcaggt
     8701 aatgagacgt tcaaaaccat ttcatttaat acgctaaaaa aagataaaat gcaaagccaa
     8761 attaaaattt ggttgtgtcg taaattcgat tgtgaatagg atgtattcac atttcaccct
     8821 ccaataatga gggcagacgt agtttatagg gttaatgata cgcttccctc ttttaattga
     8881 accctgttac attcattaca cttcataatt aattcctcct aaacttgatt aaaacatttt
     8941 accacatata aactaagttt taaattcagt atttcatcac ttatacaaca atatggcccg
     9001 tttgttgaac tactctttaa taaaataatt tttccgttcc caattccaca ttgcaataat
     9061 agaaaatcca tcttcatcgg ctttttcgtc atcatctgta tgaatcaaat cgccttcttc
     9121 tgtgtcatca aggtttaatt ttttatgtat ttcttttaac aaaccaccat aggagattaa
     9181 ccttttacgg tgtaaacctt cctccaaatc agacaaacgt ttcaaattct tttcttcatc
     9241 atcggtcata aaatccgtat cctttacagg atattttgca gtttcgtcaa ttgccgattg
     9301 tatatccgat ttatatttat ttttcggtcg aatcatttga acttttacat ttggatcata
     9361 gtctaatttc attgcctttt tccaaaattg aatccattgt ttttgattca cgtagttttc
     9421 tgtattctta aaataagttg gttccacaca taccaataca tgcatgtgct gattataaga
     9481 attatcttta ttatttattg tcacttccgt tgcacgcata aaaccaacaa gatttttatt
     9541 aattttttta tattgcatca ttcggcgaaa tccttgagcc atatctgaca aactcttatt
     9601 taattcttcg ccatcataaa catttttaac tgttaatgtg agaaacaacc aacgaactgt
     9661 tggcttttgt ttaataactt cagcaacaac cttttgtgac tgaatgccat gtttcattgc
     9721 tctcctccag ttgcacattg gacaaagcct ggatttacaa aaccacactc gatacaactt
     9781 tctttcgcct gtttcacgat tttgtttata ctctaatatt tcagcacaat cttttactct
     9841 ttcagccttt ttaaattcaa gaatatgcag aagttcaaag taatcaacat tagcgatttt
     9901 cttttctctc catggtctca cttttccact ttttgtcttg tccactaaaa cccttgattt
     9961 ttcatctgaa taaatgctac tattaggaca cat
//
