LOCUS Exported 5210 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRG02 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5210) TITLE Nematode antibiotic selection (G418) JOURNAL Unpublished REFERENCE 2 (bases 1 to 5210) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5210 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(37..55) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(76..92) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(76..92) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(89..111) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 100..116 /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(124..154) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 169..190 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(307..324) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(478..1066) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(558..577) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1237..2097) /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 1860..1879 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2098..2202) /gene="bla" /label=AmpR promoter rep_origin 2229..2685 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2316..2335) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2526..2547 /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2811..2833 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2825..2842 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2826..2842 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2849..2868 /label=T7 /note="T7 promoter, forward primer" promoter 2849..2867 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 2892..2908 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2893..2909 /label=KS primer /note="common sequencing primer, one of multiple similar variants" CDS 3712..4506 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3766..3785) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4376..4395 /label=Neo-F /note="Neomycin resistance gene, forward primer" ORIGIN 1 gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt taatttcgag 61 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc 121 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta 181 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca 241 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc 301 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 361 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 421 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 481 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 541 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 601 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 661 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 721 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 781 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 841 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 901 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt 961 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 1021 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 1081 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 1141 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 1201 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 1261 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 1321 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 1381 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 1441 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 1501 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat 1561 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 1621 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 1681 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 1741 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 1801 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga 1861 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 1921 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 1981 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 2041 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 2101 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 2161 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 2221 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag 2281 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt 2341 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt 2401 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg 2461 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt 2521 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 2581 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaaatgag ctgatttaac 2641 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag 2701 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 2761 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 2821 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattgggtac 2881 cgggcccccc ctcgaggtcg acggtatcga taagcttatt tctaaatttt agtttaataa 2941 aaaaatttta aaataatacg agttttgaga ctaactttca ataaaaacca aaaaattgct 3001 tcaaaaaaga gctcgcgccg atttacgata aacacagcgc gctacgacac tccgaaaacc 3061 gcgccgcacc attacattct ccgccccacc gcacggtttg cggaaggcga tgaaattgct 3121 gggcctaaaa ttttaaccct ttttttcccg tttttcaact gttttcccga ttttttcgat 3181 atttttttca ggtttttagt tagaaaagta tataaaaatt gcattatttg tagttttcgc 3241 tcgtaatttg tgatagaaaa cggccaattt gcctagattt gtgattctac caagtggaat 3301 ttttgggaaa atatcaatat tccacgattt ttattcaaat cataacgttt cgaaattatc 3361 agcttaaata aaactaaaat aaataataaa atactaaaaa ccctgaattt ctcggttctc 3421 ggttcagctt gagttcgttc tatacacgtt atcgattttt gtcatttcgt ttaatcatgg 3481 gttttgattc attttaaata tgaaatttga tttattcctc gtttcgaagt tggtttgtct 3541 tgtccggccg gtaaacaaat gttattaaaa aaaataataa atttcgaaat ttttcgtaat 3601 gttccgcaat cgtgtttttc cgtttatctc gtcttccctt ccttcaaata tactatttat 3661 tttctcaatt cacatatttt cagccagccc tgctcaacaa gtggaataaa aatgattgaa 3721 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 3781 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 3841 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 3901 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3961 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 4021 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 4081 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 4141 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 4201 gggctcgcac cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 4261 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 4321 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 4381 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 4441 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 4501 ttctgagtcc aattactctt caacatccct acatgctctt tctccctgtg ctcccacccc 4561 ctatttttgt tattatcaaa aaacttctct taatttcttt gttttttagc ttcttttaag 4621 ccacctctaa caatgaaatt gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc 4681 gaaaaaaatt gtgctccctc cccccattaa taataattct atcccaaaat ctacacaatg 4741 ttctgtgtac acttcttatg ttttttactt ctgataaatt tttttgaaac atcatagaaa 4801 aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat 4861 ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg 4921 agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc 4981 tttttggggt ttcccctatt gtttgtcaag atttcgagga cggcgttttt cttgctaaaa 5041 tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg tttgaggctc 5101 agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg gggtttttgc 5161 cttaaatgac agaatacatt cccaatatac caaacataac tgtttctaga //