LOCUS       Exported               13152 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Gateway adapted binary vector pCGTNG, complete sequence.
ACCESSION   DQ370425
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13152)
  AUTHORS   Zhang C, Galbraith DW.
  TITLE     A Gateway adapted Agrobacterium binary vector series for plant 
            biological studies
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13152)
  AUTHORS   Zhang C, Galbraith DW.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JAN-2006) Department of Plant Sciences, University of 
            Arizona, 303 Forbes Building, Tucson, AZ 85721-0036, USA
REFERENCE   3  (bases 1 to 13152)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13152
                     /organism="Gateway adapted binary vector pCGTNG"
                     /mol_type="other DNA"
                     /db_xref="taxon:370584"
     misc_feature    1..897
                     /label=derived from plasmid pCsGFPBT
                     /note="derived from plasmid pCsGFPBT"
     regulatory      1..758
                     /regulatory_class="promoter"
                     /note="cauliflower mosaic virus 35S promoter; duplicated"
     promoter        697..743
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     5UTR           759..893
                     /note="translation leader sequence; 5UTR from tobacco etch
                     virus"
     misc_feature    898..2608
                     /label=Invitrogen attR cassette (reading frame A)
                     /note="Invitrogen attR cassette (reading frame A)"
     protein_bind    901..1025
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1050..1080
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1134..1793
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2135..2440
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2481..2605)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    2609..13152
                     /label=derived from plasmid pCsGFPBT
                     /note="derived from plasmid pCsGFPBT"
     CDS             2610..3338
                     /codon_start=1
                     /product="sGFP"
                     /label=sGFP
                     /note="modified GFP by the Sheen Lab with enhanced 
                     fluorescence"
                     /protein_id="ABD16418.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYKGIL"
     CDS             2610..3326
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     regulatory      3339..3550
                     /regulatory_class="polyA_signal_sequence"
                     /note="cauliflower mosaic virus 35S poly(A) signal"
     primer_bind     complement(3572..3588)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4181..4433
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      4184..4436
                     /regulatory_class="polyA_signal_sequence"
                     /note="nopaline synthase poly(A) signal"
     misc_feature    4455..4479
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     repeat_region   4474..4499
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"
     misc_feature    complement(5540..6540)
                     /label=STA region from pVS1 plasmid
                     /note="STA region from pVS1 plasmid"
     CDS             5779..6408
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6842..7909
                     /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      complement(7133..8133)
                     /direction=LEFT
                     /note="pVS1-REP; replication origin from pVS1"
     rep_origin      7975..8169
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    8513..8653
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(8543..8803)
                     /label=bom site from pBR322
                     /note="bom site from pBR322"
     rep_origin      complement(8839..9427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(8943..9223)
                     /direction=LEFT
                     /label=pBR322 replication origin
                     /note="pBR322 replication origin"
     CDS             complement(9514..10308)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="aadA (kanamycin resistance) amplified from pIG121Hm"
                     /protein_id="ABD16419.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     repeat_region   10733..10758
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    10733..10757
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      complement(10825..11033)
                     /regulatory_class="polyA_signal_sequence"
                     /note="cauliflower mosaic virus 35S poly(A) signal"
     polyA_signal    10835..11009
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(11049..12074)
                     /codon_start=1
                     /product="hygromycin phosphotransferase"
                     /label=hygromycin phosphotransferase
                     /note="hptII (hygromycin resistance)"
                     /protein_id="ABD16420.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     regulatory      complement(12110..12890)
                     /regulatory_class="promoter"
                     /note="CaMV35S2; CaMV 35S promoter; duplicated"
     promoter        complement(12142..12819)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    13010..13031
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      13038..13092
                     /regulatory_class="promoter"
                     /note="lacZ promoter"
     promoter        13046..13076
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13084..13100
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     13108..13124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"