LOCUS       62056_17465        5672 bp DNA     circular SYN 19-MAY-2020
DEFINITION  Cloning vector pMT405, complete sequence.
ACCESSION   MN991273
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5672)
  AUTHORS   Taketani M, Zhang J, Zhang S, Triassi AJ, Huang YJ, Griffith LG, 
            Voigt CA.
  TITLE     Genetic circuit design automation for the gut resident species 
            Bacteroides thetaiotaomicron
  JOURNAL   Nat. Biotechnol. (2020) In press
  PUBMED    32231334
REFERENCE   2  (bases 1 to 5672)
  AUTHORS   Taketani M, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JAN-2020) Biological Engineering, MIT, Synthetic 
            Biology Center 500 Technology Square NE47-140, Cambridge, MA 02139, 
            USA
REFERENCE   3  (bases 1 to 5672)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Biotechnol. (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JAN-2020) Biological Engineering, MIT, Synthetic Biology Center 
            500 Technology Square NE47-140, Cambridge, MA 02139, USA"
FEATURES             Location/Qualifiers
     source          1..5672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(156..890)
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="QJT41631.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGSIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     CDS             1378..2235
                     /label=AmpR
                     /note="beta-lactamase"
     oriT            complement(2369..2478)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2649..3037
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     regulatory      3078..3107
                     /label=tL17
                     /note="tL17"
                     /regulatory_class="terminator"
     misc_feature    3204..3272
                     /label=LacZ alpha
                     /note="LacZ alpha"
     primer_bind     3344..3360
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3361..3417
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3430..3446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3454..3470)
                     /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)."
     promoter        complement(3478..3508)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3523..3544)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     regulatory      3544..3564
                     /label=lacO3
                     /note="lacO3"
                     /regulatory_class="other"
     regulatory      3656..3716
                     /label=L3S2P21
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     misc_recomb     3960..4289
                     /label=attN1 region
                     /note="attN1 region"
     misc_feature    4087..4100
                     /label=CC Region
                     /note="CC Region"
     misc_difference 4099
                     /replace="g"
                     /label=mutated for higher integration efficiency
                     /note="mutated for higher integration efficiency"
     CDS             4305..5642
                     /codon_start=1
                     /transl_table=11
                     /product="IntN1"
                     /label=IntN1
                     /note="NBU1 integrase"
                     /protein_id="QJT41633.1"
                     /translation="MKVTFIIKKAAKRYDTESMATIYVRFRNGRQLDSVAPTQLAINPN
                     LWDDKDECVKTKAVCNEEMRTHINEEIRQLKTYIEKVYQQEKEAIDKEWLKTTLDKFYH
                     PEKYFLPEEVVIKPTIGELFDEFLNKHPLSEVRKKNFRVVKRALLRYELYVRATKRGQK
                     GFILDVDLVTPDTLRDMWDFFQNEYQYYELYPSIYEAIPEKRTPQPRSKNTLIDCFSRI
                     RTFFLWCFDNKRTTNRPFDKFPIEECTYGTPYYITLEERDRIFNADLSATPQLAIQRDI
                     FIFQTLIGCRVSDLYRMTKLNVVNEAIEYIPKKTKEGNPVTVRVPLNDKAKEILERYKE
                     YEGKLLPFISEQKYNDAIKKIFKLAGVDRIVTILDPLTHNEIKRPIYEVASSHLARRTF
                     IGNIYKKVKDPNLVSALSGHKEGSKAFRRYRDIDEEMKKDLVKLLD"
ORIGIN
        1 agagtagcga tattaaaaga atccgatgag aaaagactaa tatttatcta tccattcagt
       61 ttgatttttc aggactttac atcgtcctga aagtatttgt tggtaccggt accgaggacg
      121 cgtaaacatt tacagttgca tgtggcctat tgtttttagc cgttaaatat tttataacta
      181 ttaaatagcg atacaaattg ttcgaaacta atattgttta tatcatatat tctcgcatgt
      241 tttaaagctt tattaaattg attttttgta aacagttttt cgtactcttt gttaacccat
      301 ttcattacaa aagtttcata tttttttctc tctttaaatg ccatttttgc tggctttctt
      361 tttaatacaa ttaatgtgct atccacttta ggttttggat ggaaataata cctaggaatt
      421 tttgctaata tagaaatatc tacctctgcc attaacagca atgctagtga tctgtttgta
      481 tctaataaca ttttagcaaa accatattcc actattaaat aacttattgt ggctgaactt
      541 tcaaaaacaa tttttcgaat tatatttgtg cttatgttgt aaggtatgct gccaaatatt
      601 ttatatggat tgtggctagg aaatgtaaat ttcagtatat catcatttac tatttgatag
      661 ttaggataat ttaagagctt attacgagtt acctcacata atttagaatc aatttctatc
      721 gccgttacaa aattacatct ctttaccaat ccagcagtaa aatgaccttt ccctgcacct
      781 atttcaaaga tgttatcttt ttcatctaaa cttatgcaat tcattatttt ttctatgtga
      841 tattttgaag taataaaatt ttgactatct tttatattta ctttgttcat tataacctct
      901 ccttaattta ttgcatctct tttcgaatat ttatgttttt tgagaaaaga acgtactcat
      961 ggttcatccc gatatgcgta tcggtctgta tatcagcaac tttctatgtg tttcaactac
     1021 aatagtcatc tattctcatc tttctgagtc caccccctgc aaagcccctc tttacgacat
     1081 aaaaattcgg tcggaaaagg tatgcaaaag atgtttctct ctttaagaga aactcttcgg
     1141 gatgcaaaaa tatgaaaata actccaattc accaaattat atagcgactt ttttacaaaa
     1201 tgctaaaatt tgttgatttc cgtcaagcaa ttgttgagca aaaatgtctt ttacgataaa
     1261 atgatacctc aatatcaact gtttagcaaa acgatatttc tcttaaagag agaaacacct
     1321 ttttgttcac caatccccga cttttaatcc cgcggccatg attgaaaaag gaagagtatg
     1381 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1441 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1501 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     1561 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1621 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1681 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     1741 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1801 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1861 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1921 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1981 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2041 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtcacgc
     2101 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2161 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2221 ctgattaagc attggtaact gtcagaccaa gtttactcat aacgcgtcaa ttcgaggggg
     2281 atcaattccg tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc
     2341 ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg
     2401 agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct
     2461 acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc
     2521 ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata
     2581 atgaccccga agcagggtta tgcagcggaa aacggaattg atccggccac gatgcgtccg
     2641 gcgtagagga tctgaagatc agcagttcaa cctgttgata gtacgtacta agctctcatg
     2701 tttcacgtac taagctctca tgtttaacgt actaagctct catgtttaac gaactaaacc
     2761 ctcatggcta acgtactaag ctctcatggc taacgtacta agctctcatg tttcacgtac
     2821 taagctctca tgtttgaaca ataaaattaa tataaatcag caacttaaat agcctctaag
     2881 gttttaagtt ttataagaaa aaaaagaata tataaggctt ttaaagcttt taaggtttaa
     2941 cggttgtgga caacaagcca gggatgtaac gcactgagaa gcccttagag cctctcaaag
     3001 caattttgag tgacacagga acacttaacg gctgacatgg gaattcccct ccaccgcggt
     3061 gtattaatgc ggctgcaatt tttttgggcg gggccgccca aaaaaatcct agcaccctgc
     3121 agggtctccc gcttctgtct tctgcaccat atgcggtgtg aaataccgca cagatgcgta
     3181 aggagaaaat accgcatcag gcgccattag ccattcaggc tgcgcaactg ttgggaaggg
     3241 cgatcggtgt gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     3301 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     3361 gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc
     3421 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3481 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3541 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3601 ttaatgaatc ggccaacgcg cggggaagac agggtctccc aatgaatggg ctttcctcgg
     3661 taccaaattc cagaaaagag gcctcccgaa aggggggcct tttttcgttt tggtccctaa
     3721 ttgcctatct tccagtgatg gaacagcatt tgtgcattgg ctgcaacaat cagccttact
     3781 tgtgcctgtt ctatttccga accgaccgct tgtatgaatc catcaaaatt cgttttctct
     3841 atgttggatt ccttgttgct catattgtga tgataatttc tacaaatata gtcattggta
     3901 actatctatg aaactgtttg atacttttac tcatcggatt cttttaatat cgctactctc
     3961 tacgttcaac caaaagaaat aatgacttac tgctatattt tttgcacgtg tggggaaaat
     4021 gtggggaaaa ttcaagcaaa agaaaaagct aagtattgaa ctatcaaata cttagctttc
     4081 tttcttgtac ccagaccccg catttgaaat aattaaagtg gggaaaatgt gggtaaaaag
     4141 aaaaatgcgg aaaaacgcca caattacact gtatttcaat atgttataat cctattaaat
     4201 tttaatccaa gtttaatcga attgcaaaat atttagcaga tgtggggaaa atgctgggga
     4261 aaatatttat atttgcagca gagtaaaatt caacctgaaa gtctatgaaa gtaaccttta
     4321 tcattaaaaa agcagccaaa cgatatgata cagaatccat ggctacaatc tatgtccgtt
     4381 ttagaaacgg aaggcagtta gactccgttg ctcctactca gttagccatc aatcccaatc
     4441 tatgggatga taaagacgaa tgtgtaaaaa cgaaagctgt ctgcaatgaa gaaatgcgta
     4501 cccatataaa tgaagagata cgccagttga aaacctatat cgagaaggta tatcaacaag
     4561 aaaaggaagc aatagacaaa gaatggctaa aaacaacact tgataaattt taccatcctg
     4621 aaaaatattt tttgccggag gaagtggtta tcaagcctac cattggagaa ctattcgatg
     4681 aatttctaaa caagcaccct ttgtcggaag tacgaaagaa aaatttccgg gttgtcaaaa
     4741 gagccttact gcgttatgaa ctatatgtaa gggctacaaa gagaggacaa aagggcttta
     4801 tccttgatgt ggatttggta acacctgaca cgcttcggga tatgtgggat ttctttcaga
     4861 acgaatacca gtattatgaa ctttacccga gcatttatga agccattccc gaaaagagga
     4921 caccacagcc cagaagcaaa aacacgctga tagactgttt ttcaagaata cgcacattct
     4981 tcctgtggtg cttcgataac aaacgcacca caaacagacc tttcgacaag tttccgatag
     5041 aggagtgtac atatggtaca ccttattata taacactcga agaaagggac aggattttta
     5101 atgcagacct ttctgccacc ccacaactgg caatacagag ggatatattc atatttcaga
     5161 cactgatagg atgcagggtg agcgacctgt accgaatgac caaactaaat gtggtcaatg
     5221 aagccataga atatattccc aagaaaacca aagaggggaa tccggttacg gtacgtgttc
     5281 cacttaacga caaagcgaaa gaaatccttg aacgctacaa agaatatgag ggaaaactgt
     5341 tgccgttcat atccgagcaa aagtacaatg atgccataaa aaagatattc aaattagctg
     5401 gagttgaccg catcgtaaca atcttagacc cgttgacgca caacgaaatc aaacgaccta
     5461 tttatgaagt ggcaagcagc catctggcaa gacgtacgtt tatcggcaat atctataaaa
     5521 aagtgaaaga cccgaacctt gtttccgcac tgtcgggaca caaggaggga agcaaagctt
     5581 tcagacgata cagggatatt gacgaagaaa tgaagaaaga ccttgtaaaa ctattagact
     5641 gataaaagta tcaaacagtt tcatagatag tt
//
