LOCUS Exported 4205 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Provides the bar gene for resistance to basta fused to the A. tumefaciens Nopaline synthase promoter and terminator. Cloned as a level 0 GB part ready to be transferred to a GB level1 plant expression vector.. ACCESSION . VERSION . KEYWORDS pTNos:BASTA:PNos (GB0023) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4205) AUTHORS Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D TITLE GoldenBraid 2.0: a comprehensive DNA assembly framework for plant synthetic biology. JOURNAL Plant Physiol. 2013 Jul;162(3):1618-31. PUBMED 23669743 REFERENCE 2 (bases 1 to 4205) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4205 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 28..280 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS complement(300..851) /codon_start=1 /gene="Streptomyces hygroscopicus bar" /product="phosphinothricin acetyltransferase" /label=BlpR /note="confers resistance to bialophos or phosphinothricin" /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF WQLDFSLPVPPRPVLPVTEI" promoter complement(893..1072) /label=NOS promoter /note="nopaline synthase promoter" promoter complement(1256..1274) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1257..1274) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1292..1308) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1292..1308) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1305..1327) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1316..1332 /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)." promoter complement(1340..1370) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1385..1406 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1523..1540) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1694..2282) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1774..1793) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2453..3313) /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 3076..3095 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3314..3418) /gene="bla" /label=AmpR promoter rep_origin complement(3496..3951) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3633..3654) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3845..3864 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4077..4099 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4091..4108 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4092..4108 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 4115..4134 /label=T7 /note="T7 promoter, forward primer" promoter 4115..4133 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 4164..4173 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 4168..4177 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 ctcgggagat cagcttgcat gccggtcgat ctagtaacat agatgacacc gcgcgcgata 61 atttatccta gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg 121 ggactctaat cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta 181 catgcttaac gtaattcaac agaaattata tgataatcat cccaagaccg gcaacaggat 241 tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc tgcttgactc taggggtcat 301 cagatttcgg tgacgggcag gaccggacgg ggcggcaccg gcaggctgaa gtccagctgc 361 cagaaaccca cgtcatgcca gttcccgtgc ttgaagccgg ccgcccgcag catgccgcgg 421 ggggcatatc cgagcgcctc gtgcatgcgc acgctcgggt cgttgggcag cccgatgaca 481 gcgaccacgc tcttgaagcc ctgtgcctcc agggacttca gcaggtgggt gtagagcgtg 541 gagcccagtc ccgtccgctg gtggcggggg gacacgtaca cggtcgactc ggccgtccag 601 tcgtaggcgt tgcgtgcctt ccagggaccc gcgtaggcta tgccggcgac ctcgccgtcc 661 acctcggcga cgagccaggg atagcgctcc cgcagacgga cgaggtcgtc cgtccactcc 721 tgcggttcct gcggctcggt acggaagttg accgtgcttg tctcgatgta gtggttgacg 781 atggtgcaga ccgccggcat gtccgcctcg gtggcacggc ggatgtcggc cgggcgtcgt 841 tctgggctca tggtagatcc cctcgatcga gttgagagtg aatatgagac tctaattgga 901 taccgagggg aatttatgga acgtcagtgg agcatttttg acaagaaata tttgctagct 961 gatagtgacc ttaggcgact tttgaacgcg caataatggt ttctgacgta tgtgcttagc 1021 tcattaaact ccagaaaccc gcggctcagt ggctccttca acgttgcggt tctgtcagtt 1081 ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt cataacgtga ctcccttaat 1141 tctcatgtat gatacgctcg agaccactca tcgcccatca ctagtgaatt cgcggccgcc 1201 tgcaggtcga ccatatggga gagctcccaa cgcgttggat gcatagcttg agtattctat 1261 agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt 1321 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg 1381 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg 1441 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 1501 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 1561 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 1621 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 1681 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 1741 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 1801 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 1861 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 1921 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 1981 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 2041 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 2101 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 2161 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 2221 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 2281 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 2341 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 2401 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 2461 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 2521 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 2581 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 2641 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 2701 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 2761 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 2821 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 2881 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 2941 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 3001 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 3061 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 3121 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 3181 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 3241 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 3301 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 3361 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 3421 catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga 3481 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt 3541 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa 3601 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa 3661 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac 3721 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga 3781 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa 3841 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc 3901 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc 3961 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca 4021 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca 4081 gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt 4141 gggcccgacg tcgcatgctc ccggccgcca tggcggccgc gggaattcga tgggcgatga 4201 gtggt //