LOCUS Exported 6405 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pPD162.21 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6405) AUTHORS Zhang H, Fire AZ TITLE Cell autonomous specification of temporal identity by Caenorhabditis elegans microRNA lin-4. JOURNAL Dev Biol. 2010 Aug 15. 344(2):603-10. PUBMED 20493184 REFERENCE 2 (bases 1 to 6405) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6405 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(3979..4001) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4101..4120 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(4198..4216) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4284..4388 /gene="bla" /label=AmpR promoter CDS 4389..5249 /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(4607..4626) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5420..6008 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5909..5928 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6162..6179 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6296..6317 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6332..6362 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6370..6386 /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 6375..6397 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6394..5 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6394..5 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 atgaccatga ttacgccaag cttgggctgc aggtcggcta taataagttc ttgaataaaa 61 taattttccc gacaaaacat gagtatttct ttcgaaaata aaagtgcagg ctaattagag 121 attattctgt aattaactgc ataatttgtc acgtgccata gttttacatt ccactacgtc 181 atagttctta aaatactaat ctcctgaaaa tagaagtagg tgaagaaagt ttaattatca 241 gttctaaaat gacaattgat ctttggaata tgttctgaaa ctaccgatca ttgaacagat 301 gctatttgaa tgatatagaa ttgtatattt gcaatttctg aaacgcgttc ttaaaggcac 361 acagattaat tcaaaagggt ctggccgcaa aaaggtttat ggtggccgat tttgagtttt 421 gtgtgtgatt gctttttcac aatcagtgtt ttcaggatta tgtgatgaac tagatcttca 481 agtttcgtta catttcatat gttttcggaa ctcacgaagt acatattggg tattgtgctc 541 aaaaaattca gcaatcagct tcgctccgct gactttagaa cccaaaaaaa tagtatggcc 601 aaactgactg tgttacgatc atttcaattt ttcaatacat atttaagatt tctaagagta 661 agaaggtcaa aaactgttct ggaatacata tatatttttc aggttacaat tagtcaaaaa 721 gtgcactgaa atatacgttt taatttcacg aataacccaa ttagttcaat gtatttttgg 781 tcaaccaacg ttaaagtttg gcttccaacc aattatcatt tctgatcaac cacaatgttt 841 tttctttatc tgcaagttaa ttttttattt ttatccagat gtttggcata tttttcaatt 901 cttcactagc gcccacttct tgcacttccg gcgccctgaa tctaatgcat ctgttgcaag 961 aattgaaaga ccaatcaaca cattgttttc ttcacgagat actgaagaaa atgaataaaa 1021 acagagaaaa agagccatgt gattagtgac aactgttgct aacagatacc atagcttgga 1081 cttggtacgt gatggcaacg tatgggtcaa caaaaatgat tgcagagggg gtgcaaaaca 1141 gtcaagtcga gaaaatatga aaaacagaaa acaaagaaca gaaaaatggg tttgagagtc 1201 agtataattt ataaaagaaa aattgtacat agaaattaac catttttgta gaagaagtta 1261 tttttcaagc atcgttaaaa attattcaaa gcaccttatt tcatatttaa ttttaaacat 1321 ggttaaatga acaacacggt gcgcaatcag gaaaacttga aatctgaaac tgttgttgtg 1381 atcttcttcg caactgttca gatagcacta gtgtaatgtt aagagtgcgc gaatataatg 1441 gaatataatg gatcacacct cctgccatca ggtaaacgtc tctgttatca catatttcca 1501 actattaaat ttttaccttt tacagtttta catttttttg aaaaaagtaa ctttttgtct 1561 tcaaaatccc tgacgaaaat atcaaatatt ttaatcgaga ctgcagagga accgattgat 1621 gatttggaaa atccagcttt acctgtgtaa gaactgaaaa gtttcataac cctagggtat 1681 tcccagttac attccccact ggctaacaat agcacccagt ttttcatcac ctttcttcaa 1741 atttctcggc gatttgttaa aaacaaaatt tgtgtccctt ctctgatatc tctatgtctc 1801 taaacacaag ttcatcggaa aacgaaggag ggtaggtgtt ggttgggctc ccgaagtgaa 1861 aatagaagag caagaataga atattagaga gagagtgcag agagggcggg atagctcccg 1921 ggattccgtt ttcttcttct ttatcttcaa cgatgatgtg tgtgcgtgtt gtatagattc 1981 tgttgctccc ccacaactcg ctccgaaggc tcaatacaat tcaattgata ttggaggaga 2041 gcctaccgga gtgggaggat aagaagaaac ataagaagaa gaagaagaag aagcatgctt 2101 ctggtttttg atgctatgaa aacggcacaa aaagatgatt gaggtccctt ttcaatacct 2161 tctctcatct ttcaaatccc attgaaacct aaaacttctc accacgcttt accattgttc 2221 tccaaaaact tatagcaatg tctataactt ttttatctct gaaaagcagt gttccatttt 2281 tctttttcct attttatttc aattgtttct cacatttcgt ttggattctt tgcttgtcaa 2341 ccagcttctt cttccacttt taccgtctaa ttttcagggc agggagccat caaacccacg 2401 accactagat ccatctagag gatccccggg attggccaaa ggacccaaag gtatgtttcg 2461 aatgatacta acataacata gaacattttc aggaggaccc ttggctagcg tcgacggtac 2521 ctcgagaaaa aatggcactc ctgcaatcac gtctcctgct atccgcacct cgtcgtgccg 2581 ctgcaactgc aagagcttca tccgtaccgg tcgccaccat gaggtcttcc aagaatgtta 2641 tcaaggagtt catgaggttt aaggttcgca tggaaggaac ggtcaatggg cacgagtttg 2701 aaatagaagg cgaaggagag gggaggccat acgaaggcca caataccgta aagcttaagg 2761 taaccaaggg gggacctttg ccatttgctt gggatatttt gtcaccacaa tttcagtatg 2821 gaagcaaggt atatgtcaag caccctgccg acataccaga ctataaaaag ctgtcatttc 2881 ctgaaggatt tgtaagttta aacggggtgt tactaactaa caaataacat ttaaattttc 2941 agaaatggga aagggtcatg aactttgaag acggtggcgt cgctactgtc acccaagatt 3001 ccagcttgca agatggctgt ttcatctaca aggtcaagtt cattggcgtg aactttcctt 3061 ccgatggacc tgttatgcaa aagaagacaa tgggctggga agccagcact gagcgtttgt 3121 atcctcgtga tggcgtgttg aaaggagaga ttcataaggc tctgaagctg aaagacgggt 3181 aagtttaaac tatatcttac taactaactg tagagattta aattttcagt ggtcattacc 3241 tagttgaatt caaaagtatt tacatggcaa agaagcctgt gcagctacca gggtactact 3301 atgttgactc caaactggat ataacaagcc acaacgaaga ctatacaatc gttgagcagt 3361 atgaaagaac cgagggacgc caccatctgt tcctttagcg gccgcgactc tagaattcca 3421 actgagcgcc ggtcgctacc attaccaact tgtctggtgt caaaaataat aggcctacta 3481 gccggccata caagtaatcc ggatgatcga cgccaacgtc gttgaatttt caaattttaa 3541 atactgaata tttgtttttt ttcctattat ttatttattc tctttgtgtt ttttttcttg 3601 ctttctaaaa aattaattca atccaaatct aaacatgagc ggtttttttt ctctttccgt 3661 ctcccaattc gtattccgct cctctcatct gaacacaatg tgcaagttta tttatcttct 3721 cgctttcatt tcattaggac gtggggggaa ttggtggaag ggggaaacac acaaaaggat 3781 gatggaaatg aaataaggac acacaatatg caacaacatt caattcagaa atatggagga 3841 aggtttaaaa gaaaacataa aaatatatag aggaggaagg aaaactagta aaaaataagc 3901 aaagaaatta ggcgaacgat gagaattgtc ctcgcttggg ccctttcgtc tcgcgcgttt 3961 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct 4021 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg 4081 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg 4141 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcggc cttaagggcc 4201 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag 4261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 4321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 4381 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt 4441 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt 4501 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt 4561 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg 4621 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga 4681 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa 4741 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga 4801 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa 4861 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca 4921 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta 4981 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac 5041 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc 5101 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag 5161 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga 5221 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt 5281 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 5341 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 5401 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 5461 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 5521 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc 5581 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 5641 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 5701 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 5761 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa 5821 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 5881 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 5941 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 6001 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 6061 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 6121 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 6181 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat 6241 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg 6301 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt 6361 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagct //