LOCUS Exported 8593 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION destination vector with cis-golgi marker (alpha-man I 49AA) -ECFP for tagged fluorescent protein (N-ter-EYFP) of plant gene. ACCESSION . VERSION . KEYWORDS pBullet-cg-n SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8593) 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, Heazlewood1 JL TITLE The Plant Glycosyltransferase Clone Collection for Functional Genomics. JOURNAL Plant J. 2014 Jun 6. doi: 10.1111/tpj.12577. PUBMED 24905498 REFERENCE 2 (bases 1 to 8593) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8593 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 177..765 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 666..685 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 919..936 /label=L4440 /note="L4440 vector, forward primer" promoter 1406..1751 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" CDS 1949..2668 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" terminator 2687..2939 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS 3107..3901 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3161..3180) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3771..3790 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3929..3947) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4015..4119 /gene="bla" /label=AmpR promoter CDS 4120..4980 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(4338..4357) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter 5005..5349 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" primer_bind 5351..5367 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 5352..5368 /label=KS primer /note="common sequencing primer, one of multiple similar variants" CDS 5377..6093 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" protein_bind 6100..6224 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS complement(6653..6958) /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" primer_bind complement(6755..6774) /label=ccdB-fwd /note="ccdB gene, forward primer" CDS complement(7303..7962) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 7877..7896 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(7963..8065) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" protein_bind complement(8243..8367) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" ORIGIN 1 acgcgtctgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt 61 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 301 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 841 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 901 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc 961 aaaccgcctc tccccgcgcg ttggccgatt cattaatgcg gccgcgagct ccctacccct 1021 actccaaaaa tgtcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac 1081 aaagggtaat ttcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatcg 1141 aaaggacagt agaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg 1201 ctatcattca agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga 1261 gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgaca 1321 tctccactga cgtaagggat gacgcacaat cccaccccta ctccaaaaat gtcaaagata 1381 cagtctcaga agaccaaagg gctattgaga cttttcaaca aagggtaatt tcgggaaacc 1441 tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta gaaaaggaag 1501 gtggctccta caaatgccat cattgcgata aaggaaaggc tatcattcaa gatgcctctg 1561 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg 1621 ttccaaccac gtcttcaaag caagtggatt gatgtgacat ctccactgac gtaagggatg 1681 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt 1741 tggagaggac agcccagatc tttgtttggg aatggctagc gggagcagat cagtgggtag 1801 cagcagcagc aaatggaggt actgcaaccc tactagttac ttgaagcgcc caaagcgtct 1861 tgctctgctc ttcatcgttt tcgtttgtgt ctctttcgtt ttctgggacc gtcaaactga 1921 cgtctcgagg accggtcccg ggggatccat ggtgagcaag ggcgaggagc tgttcaccgg 1981 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc 2041 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac 2101 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg 2161 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga 2221 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc 2281 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt 2341 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacatca gccacaacgt 2401 ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcaaga tccgccacaa 2461 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga 2521 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga 2581 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac 2641 tctcggcatg gacgagctgt acaagtaaag cggcggcccg gctgcagatc gttcaaacat 2701 ttggcaataa agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata 2761 atttctgttg aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat 2821 gagatgggtt tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa 2881 aatatagcgc gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcc 2941 gatgataagc tgtcaaacat gagagctccc ggaattgcca gctggggcgc cctctggtaa 3001 ggttgggaag ccctgcaaag taaactggat ggctttctcg ccgccaagga tctgatggcg 3061 caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 3121 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 3181 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 3241 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 3301 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 3361 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 3421 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 3481 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 3541 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 3601 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 3661 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 3721 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 3781 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 3841 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 3901 acacgtgtta attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat 3961 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga 4021 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 4081 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 4141 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 4201 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 4261 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 4321 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 4381 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 4441 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 4501 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 4561 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 4621 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 4681 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 4741 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 4801 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 4861 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 4921 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 4981 gttaacagaa gaccagaggg ctattgagac ttttcaacaa agggtaatat cgggaaacct 5041 cctcggattc cattgcccag ctatctgtca cttcatcgaa aggacagtag aaaaggaaga 5101 tggcttctac aaatgccatc attgcgataa aggaaaggct atcgttcaag atgcctctac 5161 cgacagtggt cccaaagatg gacccccacc cacgaggaac atcgtggaaa aagaagacgt 5221 tccaaccacg tcttcaaagc aagtggattg atgtgatatc tccactgacg taagggatga 5281 cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt catttcattt 5341 ggagaggacc tcgaggtcga cggtatcgat aagcttatgg tgagcaaggg cgaggagctg 5401 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 5461 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 5521 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc 5581 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 5641 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 5701 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 5761 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 5821 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 5881 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 5941 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg 6001 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 6061 gggatcactc tcggcatgga cgagctgtac aagcatatga caagtttgta caaaaaagct 6121 gaacgagaaa cgtaaaatga tataaatatc aatatattaa attagatttt gcataaaaaa 6181 cagactacat aatactgtaa aacacaacat atccagtcac tatgaatcaa ctacttagat 6241 ggtattagtg acctgtagtc gaccgacagc cttccaaatg ttcttcgggt gatgctgcca 6301 acttagtcga ccgacagcct tccaaatgtt cttctcaaac ggaatcgtcg tatccagcct 6361 actcgctatt gtcctcaatg ccgtattaaa tcataaaaag aaataagaaa aagaggtgcg 6421 agcctctttt ttgtgtgaca aaataaaaac atctacctat tcatatacgc tagtgtcata 6481 gtcctgaaaa tcatctgcat caagaacaat ttcacaactc ttatactttt ctcttacaag 6541 tcgttcggct tcatctggat tttcagcctc tatacttact aaacgtgata aagtttctgt 6601 aatttctact gtatcgacct gcagactggc tgtgtataag ggagcctgac atttatattc 6661 cccagaacat caggttaatg gcgtttttga tgtcattttc gcggtggctg agatcagcca 6721 cttcttcccc gataacggag accggcacac tggccatatc ggtggtcatc atgcgccagc 6781 tttcatcccc gatatgcacc accgggtaaa gttcacggga gactttatct gacagcagac 6841 gtgcactggc cagggggatc accatccgtc gcccgggcgt gtcaataata tcactctgta 6901 catccacaaa cagacgataa cggctctctc ttttataggt gtaaacctta aactgcattt 6961 caccagcccc tgttctcgtc agcaaaagag ccgttcattt caataaaccg ggcgacctca 7021 gccatccctt cctgattttc cgctttccag cgttcggcac gcagacgacg ggcttcattc 7081 tgcatggttg tgcttaccag accggagata ttgacatcat atatgccttg agcaactgat 7141 agctgtcgct gtcaactgtc actgtaatac gctgcttcat agcatacctc tttttgacat 7201 acttcgggta tacatatcag tatatattct tataccgcaa aaatcagcgc gcaaatacgc 7261 atactgttat ctggctttta gtaagccgga tccacgcggc gtttacgccc cgccctgcca 7321 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacagac 7381 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt 7441 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact 7501 ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg 7561 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg 7621 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa 7681 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat 7741 acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa 7801 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg 7861 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg 7921 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc 7981 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa 8041 gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt 8101 cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg 8161 tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttagtcgact acaggtcact 8221 aataccatct aagtagttga ttcatagtga ctggatatgt tgtgttttac agtattatgt 8281 agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc 8341 tcgttcagct ttcttgtaca aagtggtctg atctagagtc cgcaaaaatc accagtctct 8401 ctctacaaat ctatctctct ctatttttct ccagaataat gtgtgagtag ttcccagata 8461 agggaattag ggttcttata gggtttcgct catgtgttga gcatataaga aacccttagt 8521 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat 8581 ccagtgaagc gct //