LOCUS       Exported                5320 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMB1.3iYRS, complete sequence.
ACCESSION   KU055485
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5320)
  AUTHORS   Tack DS, Ellefson JW, Thyer R, Wang B, Gollihar J, Forster MT, 
            Ellington AD.
  TITLE     Addicting diverse bacteria to a non-canonical amino acid
  JOURNAL   Nat. Chem. Biol. (2016) In press
REFERENCE   2  (bases 1 to 5320)
  AUTHORS   Tack DS, Ellefson JW, Thyer R, Wang B, Gollihar J, Forster MT, 
            Ellington AD.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2015) Molecular Biosciences, University of Texas, 
            2500 Speedway, Austin, TX 78660, USA
REFERENCE   3  (bases 1 to 5320)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5320
                     /organism="Cloning vector pMB1.3iYRS"
                     /mol_type="other DNA"
                     /label=3iYRS-Nap3
                     /note="3iYRS-Nap3"
                     /db_xref="taxon:1776436"
     CDS             complement(1662..2195)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamicin 3-acetyltransferase"
                     /EC_number="2.3.1.60"
                     /label=aacC1
                     /note="confers gentamicin antibiotic resistance; gentamicin
                     resistance marker"
                     /protein_id="ALV85907.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(1662..2195)
                     /gene="aacC1"
                     /label=aacC1
     promoter        complement(2527..2629)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      2853..3700
                     /gene="p15A"
     gene            2853..3700
                     /gene="p15A"
                     /label=p15A
     rep_origin      complement(3155..3700)
                     /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."
     gene            3838..4919
                     /gene="tyrS"
                     /label=tyrS
                     /note="ptyrS"
     regulatory      3838..3985
                     /regulatory_class="promoter"
                     /gene="tyrS"
     CDS             3999..4919
                     /codon_start=1
                     /gene="tyrS"
                     /product="3-iodo-tyrosine-tRNA synthetase"
                     /EC_number="6.1.1.1"
                     /label=tyrS
                     /note="engineered tyrosine-tRNA ligase; derived from 
                     Methanocaldococcus jannaschii; charges amber suppressing 
                     tRNA-Nap3 with the non-canonical amino acid 
                     3-iodo-tyrosine"
                     /protein_id="ALV85908.1"
                     /translation="MDEFEMIKRNTSEIISEEELREVLKKDEKSAYIGFEPSGKIHLGH
                     YLQIKKMIDLQNAGFDIIILLADLAAYLNQKGELDEIRKIGDYNKKVFEAMGLKAKYVY
                     GSEFQLDKDYTLNVYRLALKTTLKRARRSMELIAREDENPKVAEVIYPIMQVNTSHYLG
                     VDVAVGGMEQRKIHMLARELLPKKVVCIHNPVLTGLDGEGKMSSSKGNFIAVDDSPEEI
                     RAKIKKAYCPAGVVEGNPIMEIAKYFLEYPLTIKRPEKFGGDLTVNSYEELESLFKNKE
                     LHPMYLKNAVAEELIKILEPIRKRL"
     regulatory      5096..5144
                     /regulatory_class="promoter"
                     /gene="lpp"
     gene            5096..5144
                     /gene="lpp"
                     /label=lpp
     tRNA            5162..5238
                     /gene="nap3"
                     /product="tRNA-Tyr"
                     /label=nap3
                     /note="3-iodo-tyrosine non-canonical amino acid"
     gene            5162..5238
                     /gene="nap3"
                     /label=nap3
     regulatory      5263..5292
                     /regulatory_class="terminator"
                     /gene="rrnB"
     gene            5263..5292
                     /gene="rrnB"
                     /label=rrnB