LOCUS Exported 5437 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Luciferase reporter. ACCESSION . VERSION . KEYWORDS pSGE_975_Nhe2_EcR-mut SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5437) AUTHORS Stampfel G, Kazmar T, Frank O, Wienerroither S, Reiter F, Stark A TITLE Transcriptional regulators form diverse groups with context-dependent regulatory functions. JOURNAL Nature. 2015 Nov 9. doi: 10.1038/nature15545. PUBMED 26550828 REFERENCE 2 (bases 1 to 5437) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5437 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 39..160 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(50..69) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 104..123 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(408..425) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(579..1167) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(659..678) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1338..2198) /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 1961..1980 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2199..2303) /gene="bla" /label=AmpR promoter rep_origin 2330..2785 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2417..2436) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2627..2648 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 2916..2964 /note="synthetic polyadenylation signal" misc_feature 2978..3069 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 3018..3037 /label=RVprimer3 /note="pGL3 vector, forward primer" protein_bind 3083..3107 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(3562..3586) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 3777..3786 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3783..5435 /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(3849..3867) /label=LucNrev /note="Firefly luciferase, reverse primer" ORIGIN 1 ctagagtcgg ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg 61 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat 121 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca 181 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta 241 caaatgtggt aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag 301 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct 361 ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac 421 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 481 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 541 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 601 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 661 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 721 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 781 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 841 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 901 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 961 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 1021 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 1081 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 1141 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 1201 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 1261 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 1321 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 1381 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 1441 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 1501 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 1561 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 1621 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 1681 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 1741 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 1801 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 1861 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 1921 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 1981 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 2041 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 2101 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 2161 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 2221 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 2281 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt 2341 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 2401 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 2461 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 2521 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga 2581 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 2641 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg 2701 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 2761 aacaaaatat taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa 2821 gggcgatcgg tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt 2881 aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac 2941 atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca 3001 aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa 3061 catttctcta tcgataggta ccacaagttt gtacaaaaaa gcaggctccg aattcgccct 3121 tccagttggc ctgcgataag atctgatttg cattattggc gaaattttat tagcagcagt 3181 agcgaaagct tatctgtcga gtgcggaatg actaaagact cccaattgag caattggtgt 3241 gtacatcgag tgtaattttc gagatgctga taaatctctg agggatttgt acgagtcgga 3301 ttcagtaatt gtcccgactt gagcaattgc atccgttggt attcgctatc tgctggaatg 3361 agtgtcgccc atctcattat ataaaattgg tagtcaattt aggatgatgc ctcatgcatt 3421 tttccgtctc catattcaat tgttctaacc agtggcctag gccttccaca aattcaattt 3481 ggtgttttaa tttaagtatc ttgactgtga ttacagagtg ttgacagagt acagcaatat 3541 ttgaaaacaa gggcgaattc gacccagctt tcttgtacaa agtggtagat ctgcgatcga 3601 gctcgcccgg ggatcgagcg cagcggtata aaagggcgcg gggtggctga gagcatcagt 3661 tgtgaatgaa tgttcgagcc gagcagacgt gccgctgcct tcgttaatat cctttgaata 3721 agccaacttt gaatcacaag acgcatacca aacggcattc cggtactgtt ggtaaagcca 3781 ccatggaaga cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg 3841 gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa 3901 ttgcttttac agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt 3961 ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg 4021 tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag 4081 ttgcagttgc gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca 4141 tttcgcagcc taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc 4201 aaaaaaagct cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg 4261 gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg 4321 attttgtgcc agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg 4381 gatctactgg tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct 4441 cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg 4501 ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat 4561 ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt 4621 acaagattca aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc 4681 tgattgacaa atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct 4741 ctaaggaagt cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat 4801 atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg 4861 gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga 4921 aaacgctggg cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg 4981 gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt 5041 ctggagacat agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt 5101 ctctgattaa gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc 5161 aacaccccaa catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac 5221 ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg 5281 attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg 5341 acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca 5401 taaaggccaa gaagggcgga aagatcgccg tgtaatt //