LOCUS Exported 5731 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS L4640 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5731) AUTHORS Fire A et al TITLE Fire Lab C. Elegans Vector Kit 1999 REFERENCE 2 (bases 1 to 5731) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5731 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(1353..1381) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2129..2149 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" primer_bind complement(3171..3193) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3293..3312 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(3390..3408) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3476..3580 /gene="bla" /label=AmpR promoter CDS 3581..4441 /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(3799..3818) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4612..5200 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5101..5120 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5354..5371 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5488..5509 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5524..5554 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5562..5578 /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 5567..5589 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5586..5602 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5586..5602 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 aagcttgcat gcctgcaggt cgactctaga ggatccccag cttgcatgcc tgcaggtcga 61 ggcatttgaa ttgggggtgg tggacagtaa ctgtctgtaa taataattac tcctgaccag 121 gttgcaattc gagttttgat aagcataatt ataccttgta cattgtgggt tttgtgctgt 181 ggacgtttta ttgtgacatc cccataagct acaagaacaa aaatgaaatg taaaagtatt 241 gaaaacgtcg tacattttat atctgagtag tatcctttgc tttaaatgtc cataaaacta 301 attttataat caataaaaca acgtttgtaa atcaactgag tttacaagta gagacattga 361 gggatacttt cactatgcta aagtgataat cgaccaaata ataacttcac tttggtattt 421 attcctgtct tataaatgtt atgtatgaat taaattcata tgcatatggc tcactctgac 481 aatttttaat aatcttccag atcaatattg actaccgatg cgggtggtct tttgctttga 541 attctgctga actttacacc ccgaacagca atgtgtgctt cagcctaaaa aaaagtaagt 601 gtgttaatca gtgcccccga ttcttcattt tttgcccctc tctcccgttt cgtcggcaaa 661 agaagagaaa ataaagataa gtctcaagat aggttggtaa tcgctaaagt ggttgtgtgg 721 ataagagtag caaaatggca ggaagagcac tttgcgcgca cacactgtac tcattgttct 781 ggataaaatt ctctcgttgt ttgccgtcgg atgtctgcct ctctgcattg agccggcttc 841 ttcactatct ttagttaacc taaaatgccg tttcttttct cgtatcccca ctatcccgtt 901 gaggttctct gctctcttcg ctccctaccg ccagcgagca actatccgtg ggggcgcctt 961 gctcggaaga tgggggggaa gaaagaagat ttttgctatt tgcacttgag aaagagactt 1021 ttcctgcgtc gatggttaga gaacagtgtg cagacacttt tcagctacct agaattacaa 1081 ttggatatcc ccgcctccca atccacccac ccagggaaaa agaagggctc gccgaaaatc 1141 aaagttatct ccaggctcgc gcatcccacc gagcggttga cttctctcca ccacttttca 1201 ttttaaccct cggggtacgg gattggccaa aggacccaaa ggtatgtttc gaatgatact 1261 aacataacat agaacatttt caggaggacc cttggctagc gtcgacggta ccggtagaaa 1321 aaatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg 1381 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg 1441 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tgggtaagtt 1501 taaacatata tatactaact aaccctgatt atttaaattt tcagccaaca cttgtcacta 1561 ctttcggtta tggtcttcaa tgcttcgcca gatacccaga tcatatgaaa cagcatgact 1621 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg 1681 acgggaacta caagacacgt aagtttaaac agttcggtac taactaacca tacatattta 1741 aattttcagg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa 1801 aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata 1861 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt 1921 taaacatgat tttactaact aactaatctg atttaaattt tcagaacttc aaaattagac 1981 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg 2041 gcgatggccc tgtcctttta ccagacaacc attacctgtc ctaccaatct gccctttcga 2101 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga 2161 ttacacatgg catggatgaa ctatacaaat agcattcgta gaattgtaag tttaaacttt 2221 ctcatactaa ctaacgcgct ctatttaaat tttcagaatt ccaactgagc gccggtcgct 2281 accattacca acttgtctgg tgtcaaaaat aataggggcc gctgtcatca gagtaagttt 2341 aaactgagtt ctactaacta acgagtaata tttaaatttt cagcatctcg cgcccgtgcc 2401 tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc 2461 caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct 2521 tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa 2581 aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac 2641 acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca 2701 tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat 2761 ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag 2821 ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc 2881 ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg 2941 ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg 3001 aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt 3061 ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt 3121 cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 3181 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 3241 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca 3301 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg 3361 agaaaatacc gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa 3421 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg 3481 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 3541 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 3601 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 3661 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 3721 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 3781 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga 3841 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 3901 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 3961 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 4021 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 4081 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac 4141 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 4201 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 4261 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 4321 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 4381 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 4441 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 4501 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 4561 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 4621 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 4681 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 4741 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 4801 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 4861 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 4921 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 4981 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc 5041 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 5101 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 5161 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 5221 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 5281 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 5341 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc 5401 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg 5461 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca 5521 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt 5581 tcacacagga aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta 5641 ctaactaact attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac 5701 gatggatacg ctaacaactt ggaaatgaaa t //