LOCUS       Exported               13038 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Bi-directional promoter binary vector p2CABA, complete sequence.
ACCESSION   JQ965697
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13038)
  AUTHORS   Tindamanyire JM, Townsley B, Kiggundu A, Sinha N, Tushemereirwe W.
  TITLE     Building a bi-directional promoter binary vector from the intergenic
            region of Arabidopsis thaliana cab1 and cab2 divergent genes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13038)
  AUTHORS   Tindamanyire JM, Townsley B, Kiggundu A, Sinha N, Tushemereirwe W.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-APR-2012) National Banana Research Programme, National
            Agricultural Research Laboratories, 13 Kms Kampala-Gulu Highway, 
            Kampala, Wakiso 256, Uganda
REFERENCE   3  (bases 1 to 13038)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13038
                     /organism="Bi-directional promoter binary vector p2CABA"
                     /lab_host="Escherichia coli DB3.1"
                     /mol_type="other DNA"
                     /PCR_primers="fwd_name: cab1utr, fwd_seq: 
                     aattaatcgcgaaggccttgaggttgagtagtgcagcacaaag, rev_name: 
                     cab2utr, rev_seq: 
                     cactagtatttaaatggagctcgttgaggcggccaatgaaacttttttgtg"
                     /PCR_primers="fwd_name: ocsa, fwd_seq: 
                     acccggggatcctctagactgctgagcctcgacatgttgtcg, rev_name: ocsb,
                     rev_seq: 
                     cgtctcgcatatctcattaaagcagcctaggcttaattaatcgcgaaggcctaaagag 
                     ctccatttaaatactagtgacgtcgcatgctcccggccgcca"
                     /db_xref="taxon:1201315"
     regulatory      1..707
                     /regulatory_class="terminator"
                     /note="tOCS; octopine synthase terminator"
     terminator      1..707
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     regulatory      735..2926
                     /regulatory_class="promoter"
                     /note="Atca/b; from Arabidopsis thaliana chlorophyll a/b 
                     gene; single natural bidirectional promoter that can drive 
                     and express two different genes at the same time"
     5UTR           735..800
                     /label=from chlorophyll a/b1 (CAB1) gene
                     /note="from chlorophyll a/b1 (CAB1) gene"
     regulatory      2965..2974
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_recomb     2987..3111
                     /note="AttR1; recombination site; derived from Escherichia 
                     coli"
     protein_bind    2987..3111
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     gene            3112..4566
                     /gene="ccdB"
                     /label=ccdB
                     /note="derived from Escherichia coli"
     promoter        3136..3166
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3220..3879
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             4221..4526
                     /codon_start=1
                     /gene="ccdB"
                     /product="cell cycle death B"
                     /label=ccdB
                     /note="cytotoxic protein; cell cycle death B; cytotoxic 
                     protein derived from Escherichia coli"
                     /protein_id="AFN68288.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_recomb     4567..4691
                     /note="AttR2; recombination site; derived from Escherichia 
                     coli"
     protein_bind    complement(4567..4691)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      4704..4914
                     /regulatory_class="terminator"
                     /note="t35S; cauliflower mosaic virus 35S terminator"
     misc_feature    4916..5115
                     /note="RB; right border of Agrobacterium T-DNA"
     misc_feature    4977..5001
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6301..6930
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7364..8431
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      8497..8691
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    9035..9175
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9361..9949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10193..11125)
                     /codon_start=1
                     /gene="spR"
                     /product="spectinomycin resistance protein"
                     /label=spR
                     /note="SpR"
                     /protein_id="AFN68287.1"
                     /translation="MTCFFGVQSMPRASKQQARYAVGRCLMLWSSNDVTQQGSRPKTKL
                     NIMGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVR
                     LDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRND
                     ILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNS
                     PPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQ
                     EEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(10193..11125)
                     /gene="spR"
                     /label=spR
     CDS             complement(10193..10984)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_recomb     11274..11606
                     /note="LB; left border of Agrobacterium T-DNA"
     misc_feature    11512..11536
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      11629..11881
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(11921..12715)
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /label=kanR
                     /note="kanamycin resistance protein"
                     /protein_id="AFN68289.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(11921..12715)
                     /gene="kanR"
                     /label=kanR
     promoter        complement(12749..12928)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"