LOCUS       40924_4244        3951 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJM.773, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3951)
  AUTHORS   Meyer AJ, Segall-Shapiro TH, Voigt CA.
  TITLE     Marionette: E. coli containing 12 highly-optimized small molecule 
            sensors
  JOURNAL   bioRxivorg 285866, 1-26 (2018)
REFERENCE   2  (bases 1 to 3951)
  AUTHORS   Meyer AJ, Segall-Shapiro TH, Voigt CA.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAR-2018) Synthetic Biology Center, Department of 
            Biological Engineering, Massachusetts Institute of Technology, 77 
            Massachusetts Avenue, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 3951)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3951)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxivorg 
            285866, 1-26 (2018)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-MAR-2018) Synthetic Biology Center, Department of Biological 
            Engineering, Massachusetts Institute of Technology, 77 Massachusetts
            Avenue, Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3951
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..53
                     /label=L3S3P21
                     /note="L3S3P21"
                     /regulatory_class="terminator"
     regulatory      73..122
                     /label=PVanCC
                     /note="PVanCC"
                     /regulatory_class="promoter"
     misc_feature    73..84
                     /label=VanO operator
                     /note="VanO operator"
     regulatory      84..89
                     /regulatory_class="minus_35_signal"
     regulatory      107..112
                     /regulatory_class="minus_10_signal"
     misc_feature    111..122
                     /label=VanO operator
                     /note="VanO operator"
     regulatory      119
                     /label=+1
                     /note="+1"
                     /regulatory_class="other"
     misc_RNA        124..174
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     regulatory      206..217
                     /label=B0064
                     /note="B0064"
                     /regulatory_class="ribosome_binding_site"
     CDS             224..940
                     /label=EYFP
                     /note="enhanced YFP"
     regulatory      944..1004
                     /label=L3S2P21
                     /note="L3S2P21"
                     /regulatory_class="terminator"
     regulatory      1009..1062
                     /label=PLacIQ
                     /note="PLacIQ"
                     /regulatory_class="promoter"
     regulatory      1027..1032
                     /regulatory_class="minus_35_signal"
     regulatory      1050..1055
                     /regulatory_class="minus_10_signal"
     regulatory      1063..1091
                     /label=van2
                     /note="van2"
                     /regulatory_class="ribosome_binding_site"
     CDS             1092..1826
                     /codon_start=1
                     /product="VanR-AM"
                     /label=VanR-AM
                     /protein_id="AYJ72236.1"
                     /translation="MDMPRIKPGQRVMMALRKMIASGEIKSGERIAEIPTAAALGVSRM
                     PVRIALRSLEQEGLVVRLGARGYAARGVSSDQIRDAIEVRGVLEGFAARRLAERGMTAE
                     THARFVVLIAEGEALFAAGRLNGEDLDRYAAYNQAFHDTLVSAAGNGAVESALARNGFE
                     PFAAAGALALDLMDLSAEYEHLLAAHRQHQAVLDAVSCGDAEGAERIMRDHALAAIRNA
                     KVFEAAASAGAPLGAAWSIRAD"
     regulatory      1836..2090
                     /label=induction operon terminator region
                     /note="induction operon terminator region"
                     /regulatory_class="terminator"
     rep_origin      complement(2402..2947)
                     /direction=LEFT
                     /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             complement(3039..3851)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 ccaattattg aaggcctccc taacgggggg cctttttttg tttctggtct cccgcttaac
       61 gatcgttggc tgattggatc caattgacag ctagctcagt cctaggtacc attggatcca
      121 atagctgtca ccggatgtgc tttccggtct gatgagtccg tgaggacgaa acagcctcta
      181 caaataattt tgtttaatac tagagaaaga ggggaaatac tagatggtga gcaagggcga
      241 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      301 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      361 gttcatctgc accacaggca agctgcccgt gccctggccc accctcgtga ccaccttcgg
      421 ctacggcctg caatgcttcg cccgctaccc cgaccacatg aagctgcacg acttcttcaa
      481 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
      541 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      601 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      661 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
      721 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
      781 caccccaatc ggcgacggcc ccgtgctgct gcccgacaac cactacctta gctaccagtc
      841 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctactgg agttcgtgac
      901 cgccgccggg atcactctcg gcatggacga gctgtacaag taactcggta ccaaattcca
      961 gaaaagaggc ctcccgaaag gggggccttt tttcgttttg gtccaatggc ggcgcgccat
     1021 cgaatggtgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
     1081 gggggtgaat aatggacatg cctcgtatta aaccgggtca gcgtgttatg atggcactgc
     1141 gtaaaatgat tgcaagcggt gaaatcaaaa gtggtgaacg tattgcagaa attccgaccg
     1201 cagcagcact gggtgttagc cgtatgccgg ttcgtatcgc actgcgttca ctggaacaag
     1261 aaggtctggt tgttcgtctg ggtgcacgtg gttatgcagc ccgtggtgtt agcagcgatc
     1321 agattcgtga tgcaattgaa gttcgtggtg ttctggaagg ttttgcagca cgtcgtctgg
     1381 cagaacgtgg tatgaccgca gaaacccatg cacgttttgt tgtactgatt gcagaaggtg
     1441 aagcactgtt tgcagccggt cgcctgaatg gtgaagatct ggatcgttat gccgcatata
     1501 atcaggcatt tcatgatacc ctggttagcg cagcaggtaa tggtgcagtt gaaagcgcac
     1561 tggcacgtaa tggttttgaa ccgtttgcag cagccggtgc actggccctg gatctgatgg
     1621 acctgtctgc cgaatatgaa catctgctgg cagcacatcg tcagcatcag gcagttctgg
     1681 atgcagttag ctgtggtgat gccgaaggtg cagaacgtat tatgcgtgat catgcactgg
     1741 cagcaattcg taatgcaaaa gtttttgaag cagcagcaag cgcaggcgca ccgctgggtg
     1801 cagcatggtc aattcgtgca gattgataag gatcctaatt ggtaacgaat cagacaattg
     1861 acggctcgag ggagtagcat agggtttgca gaatccctgc ttcgtccatt tgacaggcac
     1921 attatgcatc gatgataagc tgtcaaacat gagcagatcc tctacgccag acgcatcgtg
     1981 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     2041 ggggaagatc gggctcgcca cttcgggctc atgagcaaat attttatctg aggtgcttcc
     2101 tcgctcactg actcgctgca cgaggcagac ctcagcgcta gcggagtgta tactggctta
     2161 ctatgttggc actgatgagg gtgtcagtga agtgcttcat gtggcaggag aaaaaaggct
     2221 gcaccggtgc gtcagcagaa tatgtgatac aggatatatt ccgcttcctc gctcactgac
     2281 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
     2341 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
     2401 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
     2461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
     2521 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
     2581 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
     2641 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2701 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     2761 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     2821 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     2881 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     2941 atctcaagaa gatcatctta ttaaggggtc tgacgctcag tggaacgaaa aatcaatcta
     3001 aagtatatat gagtaaactt ggtctgacag ttaccttaga aaaactcatc gagcatcaaa
     3061 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     3121 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     3181 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     3241 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     3301 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     3361 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     3421 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     3481 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     3541 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     3601 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     3661 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     3721 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     3781 tataaatcag catccatgtt ggaatttaat cgcggcctcg agcaagacgt ttcccgttga
     3841 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     3901 gatgatatat ttttatcttg tgcaatgtac atcagagatt ttgagacaca a
//
