LOCUS Exported 6056 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS M71 pGL3 Sp5l P1203 mut SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6056) TITLE Other Plasmids REFERENCE 2 (bases 1 to 6056) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6056 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 1320..1329 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1326..2978 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(1392..1410) /label=LucNrev /note="Firefly luciferase, reverse primer" polyA_signal 3019..3140 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3030..3049) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3084..3103 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3388..3405) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3559..4147) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3639..3658) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4318..5178) /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 4941..4960 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5179..5283) /gene="bla" /label=AmpR promoter rep_origin 5310..5765 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5397..5416) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5607..5628 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 5896..5944 /note="synthetic polyadenylation signal" misc_feature 5958..6049 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 5998..6017 /label=RVprimer3 /note="pGL3 vector, forward primer" ORIGIN 1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagacg ttaagcacgt ttcctacgtg 61 tcattttaat tctccatcac tgtggtatgt tttagcagac taaagcgatg catctgctgg 121 gacaaaggtc gattgttatg cgactagagg tgcgaggcaa cacaaagcag tgaggatttt 181 ctatgtatta agatgttaaa tggctttccg acttcaatgt aaattctcta agagaaaggc 241 tcaaagtgtt agatgaactt tggatgctct gcatttaatc cctttaaacg tgatgacgcc 301 cttaacttga acccggagtg tgaaaatatc acgcagggct ctttatttat aaaatcactt 361 aagagtgcga actactgaat gtgcatggag gtacatatcg cgtttattaa tgagaaaaga 421 agaatagctt aattcgatcg ttttcaaatg tcgtttagcg tcatcattaa tatgacatct 481 tttagaacga ccagcttaca actaataaca gctaaagagg caggttcaca gatgctcata 541 tggatcaaat acaggcctag tgaaagcaaa atccataacg cactttacaa aatagcttag 601 tatcacttta aacccaacat gaggcctaac aataccaaca atgtatttgc ctatagtgaa 661 tcttgaatgc cacattcatg ttacagacct attttcatta tcatttgaag tgcacgactg 721 cagtatacta caataaaata aactaatctt aaatttaaaa gtaaagattt tggagattca 781 ttgcacctaa tctattgaaa agtatttatt ttagtgtaaa aactagtgtg gcttaaataa 841 aacggtgtta attaaaataa aaatgttata aatactgatt ataacaagct cgtccgtgta 901 catagttttt aatttgtcac taaagaacga gtcgttaaac ttgtaaatat cagatacatg 961 taaaagatct ctgtcatatt aagactcttc tcacctagaa gcgctaaagg gaaggggttc 1021 tcacaggtta acatttggca aaattaaatt agaatgaaac gaaggttata atacacaccc 1081 ctcttcctaa tctcccaaag attgccctcc ccaaatattc ccccaccgta tctctcctcc 1141 attcactggc cctcctgata catacttaag agaccccgtc gagaggatct tttccctata 1201 aagagaagat gcaatccgac cgctcgagaa atgatcctca ggacttattg cgacaggccc 1261 ttcagtcgca cttcaaaagg aagagacctg aagcttggca ttccggtact gttggtaaag 1321 ccaccatgga agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag 1381 atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa 1441 caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa 1501 tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg 1561 tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg 1621 gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg 1681 gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg 1741 tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc 1801 agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat 1861 acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct 1921 ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat 1981 tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa 2041 gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg 2101 gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg 2161 attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca 2221 ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc 2281 tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag 2341 gatatgggct cactgagact acatcagcta ttctgattac acccgagggg gatgataaac 2401 cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg 2461 ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt 2521 ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac 2581 attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga 2641 agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc 2701 tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg 2761 aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg 2821 tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg 2881 tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc 2941 tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg ggcggccggc 3001 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc 3061 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta 3121 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg 3181 gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata 3241 aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg 3301 cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga 3361 caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 3421 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 3481 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 3541 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 3601 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 3661 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 3721 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 3781 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 3841 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 3901 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 3961 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 4021 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 4081 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 4141 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 4201 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 4261 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 4321 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 4381 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 4441 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 4501 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 4561 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 4621 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 4681 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 4741 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 4801 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 4861 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 4921 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 4981 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 5041 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 5101 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 5161 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 5221 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 5281 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc 5341 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc 5401 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 5461 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 5521 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 5581 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 5641 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt 5701 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac 5761 aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc 5821 ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt 5881 acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg 5941 tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta 6001 gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgata //