LOCUS Exported 4275 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pbacPuroR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4275) AUTHORS Semple JI, Biondini L, Lehner B TITLE Generating transgenic nematodes by bombardment and antibiotic selection. JOURNAL Nat Methods. 2012 Jan 30;9(2):118-9. doi: 10.1038/nmeth.1864. PUBMED 22290182 REFERENCE 2 (bases 1 to 4275) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4275 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 140..244 /gene="bla" /label=AmpR promoter CDS 245..844 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(245..264) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 741..761 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(1849..1871) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 1971..1990 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 2154..2258 /gene="bla" /label=AmpR promoter CDS 2259..3119 /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(2477..2496) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3290..3878 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3779..3798 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4032..4049 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4166..4187 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4202..4232 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4240..4256 /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 4245..4267 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4264..5 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4264..5 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 atgaccatga ttacgccaag cttgagaaaa taccgcatca ggcggcctta agggcctcgt 61 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg 121 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 181 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 241 gagtatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc 301 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc 361 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct 421 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc 481 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag 541 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa 601 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct 661 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt 721 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac 781 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc 841 ctgactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta 901 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 961 tgagttttcg ttccactgag cgtcagaccc cgtagagcta gcgtcgacgg taccatggta 1021 ttgatatctg agctccgcat cggccgctgt catcagatcg ccatctcgcg cccgtgcctc 1081 tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca 1141 ccccctattt ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt 1201 taagtcacct ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa 1261 agtcgaaaaa aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac 1321 aatgttctgt gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata 1381 gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt 1441 ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt 1501 ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt 1561 ttgctttttg gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct 1621 aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag 1681 gctcagtgga aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt 1741 ttgccttaaa tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg 1801 gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 1861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 1921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 1981 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 2041 aaaataccgc atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg 2101 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa 2161 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 2221 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 2281 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 2341 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 2401 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 2461 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 2521 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 2581 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 2641 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 2701 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 2761 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 2821 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 2881 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 2941 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 3001 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 3061 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 3121 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 3181 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 3241 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 3301 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 3361 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 3421 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 3481 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 3541 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 3601 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 3661 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg 3721 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 3781 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 3841 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 3901 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 3961 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa 4021 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc 4081 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga 4141 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg 4201 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc 4261 acacaggaaa cagct //