LOCUS       Exported                9292 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBullet-pm-n, complete sequence.
ACCESSION   KJ081779
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9292)
  AUTHORS   Lao J, Oikawa A, Bromley JR, McInerney P, Suttangkakul A, 
            Smith-Moritz AM, Plahar H, Chiu TY, Gonzalez Fernandez-Nino SM, 
            Ebert B, Yang F, Christiansen KM, Hansen SF, Stonebloom S, Adams PD,
            Ronald PC, Hillson NJ, Hadi MZ, Vega-Sanchez ME, Loque D, Scheller 
            HV, Heazlewood JL.
  TITLE     The plant glycosyltransferase clone collection for functional 
            genomics
  JOURNAL   Plant J. 79 (3), 517-529 (2014)
  PUBMED    24905498
REFERENCE   2  (bases 1 to 9292)
  AUTHORS   Lao J, Oikawa A, Bromley JR, Smith-Moritz AM, Chiu T-Y., 
            Christiansen KM, Hansen SF, Suttangkakul A, Vega-Sanchez ME, Yang F,
            Stonebloom S, Ebert B, Morrison S, McInerney P, Hadi MZ, Adams PD, 
            Ronald PC, Loque D, Scheller HV, Heazlewood JL.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JAN-2014) Joint BioEnergy Institute, Lawrence Berkeley
            National Laboratory, One Cyclotron Road, Berkeley, CA 94720, USA
REFERENCE   3  (bases 1 to 9292)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9292
                     /organism="Cloning vector pBullet-pm-n"
                     /mol_type="other DNA"
                     /db_xref="taxon:1480049"
     rep_origin      7..998
                     /note="pUC Origin Plus; ColE1 origin"
     rep_origin      177..765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      1007..1754
                     /regulatory_class="promoter"
                     /note="double 35S promoter"
     promoter        1406..1751
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             1772..3367
                     /codon_start=1
                     /product="ECFP-tagged plasma membrane intrinsic protein 2a"
                     /label=ECFP-tagged plasma membrane intrinsic protein 2a
                     /note="AT3g53420; aquaporin PIP2A; enhanced cyan 
                     fluorescent protein tag"
                     /protein_id="AHW81492.1"
                     /translation="MAKDVEAVPGEGFQTRDYQDPPPAPFIDGAELKKWSFYRAVIAEF
                     VATLLFLYITVLTVIGYKIQSDTDAGGVDCGGVGILGIAWAFGGMIFILVYCTAGISGG
                     HINPAVTFGLFLARKVSLPRALLYIIAQCLGAICGVGFVKAFQSSYYTRYGGGANSLAD
                     GYSTGTGLAAEIIGTFVLVYTVFSATDPKRSARDSHVPVLAPLPIGFAVFMVHLATIPI
                     TGTGINPARSFGAAVIYNKSKPWDDHWIFWVGPFIGAAIAAFYHQFVLRASGSKSLGSF
                     RSAANVVPGGSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKANF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
                     TAAGITLGMDELYK"
     CDS             2648..3367
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2648..3364
                     /note="ECFP; Region: enhanced cyan fluorescent protein"
     terminator      3386..3638
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      3391..3638
                     /regulatory_class="terminator"
                     /note="NOS"
     regulatory      3639..3805
                     /regulatory_class="promoter"
                     /note="kan"
     CDS             3806..4600
                     /codon_start=1
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aminoglycoside 3-phosphotransferase
                     /note="KanR"
                     /protein_id="AHW81493.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      4608..4818
                     /regulatory_class="promoter"
                     /note="AMP"
     promoter        4714..4818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4819..5679
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="AmpR"
                     /protein_id="AHW81494.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     regulatory      5685..6050
                     /regulatory_class="promoter"
                     /note="CaMV 35S"
     promoter        5704..6048
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     6051..6067
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6076..6834
                     /codon_start=1
                     /product="enhanced yellow fluorescent protein"
                     /label=enhanced yellow fluorescent protein
                     /note="EYFP"
                     /protein_id="AHW81495.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKHMTSLYKKAERET"
     CDS             6076..6792
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    6799..6923
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     6799..6823
                     /label=attR1
                     /note="attR1"
     CDS             complement(7352..7657)
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /protein_id="AHW81496.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(8002..8661)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=chloramphenicol acetyltransferase
                     /note="CAT/CamR"
                     /protein_id="AHW81497.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(8662..8764)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(8942..9066)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_recomb     9042..9066
                     /label=attR2
                     /note="attR2"
     regulatory      9131..9286
                     /regulatory_class="terminator"
                     /note="CaMV 35S"