LOCUS       62056_17470        6083 bp DNA     circular SYN 19-MAY-2020
DEFINITION  Cloning vector pMT406, complete sequence.
ACCESSION   MN991274
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6083)
  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 6083)
  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 6083)
  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..6083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(156..890)
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="QJT41634.1"
                     /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    3134..3416
                     /label=PAM3
                     /note="PAM3"
     regulatory      3377..3380
                     /label=-33 sigma factor recognitions site
                     /note="-33 sigma factor recognitions site"
                     /regulatory_class="other"
     regulatory      3403..3410
                     /label=-7 sigma factor recognitions site
                     /note="-7 sigma factor recognitions site"
                     /regulatory_class="other"
     regulatory      3417
                     /label=BT1311
                     /note="BT1311"
                     /regulatory_class="ribosome_binding_site"
     misc_RNA        3418..3468
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      3492..3534
                     /label=rpiL*
                     /note="rpiL*"
                     /regulatory_class="terminator"
     CDS             3535..4047
                     /label=Nluc
                     /note="NanoLuc(R) luciferase"
     regulatory      4067..4127
                     /label=L3S2P21
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     primer_bind     complement(4253..4273)
                     /label=pNBU1_seq.REV
                     /note="pNBU1_seq.REV"
     misc_recomb     4371..4700
                     /label=attN1 region
                     /note="attN1 region"
     misc_feature    4498..4511
                     /label=CC Region
                     /note="CC Region"
     misc_difference 4510
                     /replace="g"
                     /label=mutated for higher integration efficiency
                     /note="mutated for higher integration efficiency"
     CDS             4716..6053
                     /codon_start=1
                     /transl_table=11
                     /product="IntN1"
                     /label=IntN1
                     /note="NBU1 integrase"
                     /protein_id="QJT41637.1"
                     /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 gcttgatctg gaagaagcaa tgaaagctgc tgttaagtct ccgaatcagg
     3181 tattgttcct gacaggtgta ttcccatccg gtaaacgcgg atactttgca gttgatctga
     3241 ctcaggaata aattataaat taaggtaaga agattgtagg ataagctaat gaaatagaaa
     3301 aaggatgccg tcacacaact tgtcggcatt cttttttgtt ttattagttg aaaatatagt
     3361 gaaaactttg caacagcata gctcagcaca gattatttgt cgtaactttg cactccagct
     3421 gtcaccggat gtgctttccg gtctgatgag tccgtgagga cgaaacagcc tctacaaata
     3481 attttgttta acgcatttta aaataaaata aattatttat ttaattaaac gaatatggtt
     3541 tttactctgg aagattttgt tggcgattgg cgtcagaccg cgggttataa tttggatcaa
     3601 gtcctggaac agggtggcgt aagctctctg ttccagaacc tgggtgtgag cgtgacgccg
     3661 attcagcgca tcgttctgtc cggcgagaac ggtctgaaaa ttgatattca tgtgatcatc
     3721 ccgtacgaag gcctgagcgg tgaccaaatg ggtcaaatcg agaaaatctt taaagtcgtc
     3781 tacccagttg acgatcacca cttcaaggtt atcttgcatt acggtacgct ggtgattgat
     3841 ggtgtgaccc cgaatatgat tgactatttc ggccgtccgt atgaaggcat tgccgttttt
     3901 gacggtaaaa agatcaccgt caccggtacc ctgtggaatg gcaataagat tattgacgag
     3961 cgtctgatta acccggacgg cagcctgctg ttccgcgtga ccatcaacgg tgtcacgggt
     4021 tggcgtctgt gcgagcgcat cctggcataa caatgaatgg gctttcctcg gtaccaaatt
     4081 ccagaaaaga ggcctcccga aaggggggcc ttttttcgtt ttggtcccta attgcctatc
     4141 ttccagtgat ggaacagcat ttgtgcattg gctgcaacaa tcagccttac ttgtgcctgt
     4201 tctatttccg aaccgaccgc ttgtatgaat ccatcaaaat tcgttttctc tatgttggat
     4261 tccttgttgc tcatattgtg atgataattt ctacaaatat agtcattggt aactatctat
     4321 gaaactgttt gatactttta ctcatcggat tcttttaata tcgctactct ctacgttcaa
     4381 ccaaaagaaa taatgactta ctgctatatt ttttgcacgt gtggggaaaa tgtggggaaa
     4441 attcaagcaa aagaaaaagc taagtattga actatcaaat acttagcttt ctttcttgta
     4501 cccagacccc gcatttgaaa taattaaagt ggggaaaatg tgggtaaaaa gaaaaatgcg
     4561 gaaaaacgcc acaattacac tgtatttcaa tatgttataa tcctattaaa ttttaatcca
     4621 agtttaatcg aattgcaaaa tatttagcag atgtggggaa aatgctgggg aaaatattta
     4681 tatttgcagc agagtaaaat tcaacctgaa agtctatgaa agtaaccttt atcattaaaa
     4741 aagcagccaa acgatatgat acagaatcca tggctacaat ctatgtccgt tttagaaacg
     4801 gaaggcagtt agactccgtt gctcctactc agttagccat caatcccaat ctatgggatg
     4861 ataaagacga atgtgtaaaa acgaaagctg tctgcaatga agaaatgcgt acccatataa
     4921 atgaagagat acgccagttg aaaacctata tcgagaaggt atatcaacaa gaaaaggaag
     4981 caatagacaa agaatggcta aaaacaacac ttgataaatt ttaccatcct gaaaaatatt
     5041 ttttgccgga ggaagtggtt atcaagccta ccattggaga actattcgat gaatttctaa
     5101 acaagcaccc tttgtcggaa gtacgaaaga aaaatttccg ggttgtcaaa agagccttac
     5161 tgcgttatga actatatgta agggctacaa agagaggaca aaagggcttt atccttgatg
     5221 tggatttggt aacacctgac acgcttcggg atatgtggga tttctttcag aacgaatacc
     5281 agtattatga actttacccg agcatttatg aagccattcc cgaaaagagg acaccacagc
     5341 ccagaagcaa aaacacgctg atagactgtt tttcaagaat acgcacattc ttcctgtggt
     5401 gcttcgataa caaacgcacc acaaacagac ctttcgacaa gtttccgata gaggagtgta
     5461 catatggtac accttattat ataacactcg aagaaaggga caggattttt aatgcagacc
     5521 tttctgccac cccacaactg gcaatacaga gggatatatt catatttcag acactgatag
     5581 gatgcagggt gagcgacctg taccgaatga ccaaactaaa tgtggtcaat gaagccatag
     5641 aatatattcc caagaaaacc aaagagggga atccggttac ggtacgtgtt ccacttaacg
     5701 acaaagcgaa agaaatcctt gaacgctaca aagaatatga gggaaaactg ttgccgttca
     5761 tatccgagca aaagtacaat gatgccataa aaaagatatt caaattagct ggagttgacc
     5821 gcatcgtaac aatcttagac ccgttgacgc acaacgaaat caaacgacct atttatgaag
     5881 tggcaagcag ccatctggca agacgtacgt ttatcggcaa tatctataaa aaagtgaaag
     5941 acccgaacct tgtttccgca ctgtcgggac acaaggaggg aagcaaagct ttcagacgat
     6001 acagggatat tgacgaagaa atgaagaaag accttgtaaa actattagac tgataaaagt
     6061 atcaaacagt ttcatagata gtt
//
