LOCUS       Exported                7308 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-Z4-PmasRFP-ALCR, complete sequence.
ACCESSION   KY825159
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7308)
  AUTHORS   Pasin F, Bedoya LC, Bernabe-Orts JM, Gallo A, Simon-Mateo C, Orzaez 
            D, Garcia JA.
  TITLE     Multiple T-DNA Delivery to Plants Using Novel Mini Binary Vectors 
            with Compatible Replication Origins
  JOURNAL   ACS Synth Biol (2017) In press
  PUBMED    28657330
REFERENCE   2  (bases 1 to 7308)
  AUTHORS   Pasin F, Garcia JA.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAR-2017) Department of Plant Molecular Genetics, 
            Centro Nacional de Biotecnologia (CNB-CSIC), Darwin 3, Madrid, 
            Madrid 28049, Spain
REFERENCE   3  (bases 1 to 7308)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7308
                     /organism="Cloning vector pLX-Z4-PmasRFP-ALCR"
                     /mol_type="other DNA"
                     /db_xref="taxon:2016281"
     regulatory      27..58
                     /regulatory_class="terminator"
     misc_feature    59..83
                     /label=LB
                     /note="LB"
     misc_feature    complement(106..129)
                     /note="left border of left T-DNA; LB"
     regulatory      178..559
                     /regulatory_class="promoter"
                     /note="mas"
     promoter        179..559
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             570..3821
                     /codon_start=1
                     /product="polyprotein"
                     /label=polyprotein
                     /protein_id="ASG92170.1"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGGGGGRAEGRGSLLTCGDVEENPGPMADTRRRQNHSCDPCRKGK
                     RRCDAPENRNEANENGWVSCSNCKRWNKDCTFNWLSSQRSKAKGAAPRARTKKARTATT
                     TSEPSTSAATIPTPESDNHDAPPVINSHDALPSWTQGLLSHPGDLFDFSHSAIPANAED
                     AANVQSDAPFPWDLAIPGDFSMGQQLEKPLSPLSFQAVLLPPHSPNTDDLIRELEEQTT
                     DPDSVTDTNSVQQVAQDGSLWSDRQSPLLPENSLCMASDSTARRYARSTMTKNLMRIYH
                     DSMENALSCWLTEHNCPYSDQISYLPPKQRAEWGPNWSNRMCIRVCRLDRVSTSLRGRA
                     LSAEEDKAAARALHLAIVAFASQWTQHAQRGAGLNVPADIAADERSIRRNAWNEARHAL
                     QHTTGIPSFRVIFANIIFSLTQSVLDDDEQHGMGARLDKLLENDGAPVFLETANRQLYT
                     FRHKFARMQRRGKAFNRLPGGSVASTFAGIFETPTPSSESPQLDPVVASEEHRSTLSLM
                     FWLGIMFDTLSAAMYQRPLVVSDEDSQISSASPPRRGAETPINLDCWEPPRQVPSNQEK
                     SDVWGDLFLRTSDSLPDHESHTQISQPAARWPCTYEQAAAALSSATPVKVLLYRRVTQL
                     QTLLYRGASPARLEAAIQRTLYVYNHWTAKYQPFMQDCVANHELLPSRIQSWYVILDGH
                     WHLAAMLLADVLESIDRDSYSDINHIDLVTKLRLDNALAVSALARSSLRGQELDPGKAS
                     PMYRHFHDSLTEVAFLVEPWTVVLIHSFAKAAYILLDCLDLDGQGNALAGYLQLRQNCN
                     YCIRALQFLGRKSDMAALVAKDLERGLNGKVDSFL"
     mat_peptide     573..1280
                     /product="TagRFP-T"
     mat_peptide     1281..1355
                     /product="T2A"
     CDS             1302..1355
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     mat_peptide     1356..3818
                     /product="ALCR"
     regulatory      3824..4076
                     /regulatory_class="terminator"
                     /note="mas"
     terminator      3824..4076
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    4131..4154
                     /label=RB
                     /note="RB"
     regulatory      complement(4225..4277)
                     /regulatory_class="terminator"
     CDS             complement(4338..4871)
                     /codon_start=1
                     /gene="acc3"
                     /product="gentamicin-3-acetyltransferase"
                     /label=acc3
                     /note="AacC1"
                     /protein_id="ASG92171.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            complement(4338..4871)
                     /gene="acc3"
                     /label=acc3
     promoter        complement(5060..5088)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     misc_feature    5089..5112
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(5163..6311)
                     /codon_start=1
                     /gene="trfA"
                     /product="replication protein TrfA"
                     /label=trfA
                     /protein_id="ASG92172.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(5163..6311)
                     /gene="trfA"
                     /label=trfA
     rep_origin      6704..7235
                     /label=oriV
                     /note="incP origin of replication"