LOCUS Exported 5967 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION NanoLuc control expression vector. ACCESSION . VERSION . KEYWORDS pcDNA3.1 NL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5967) AUTHORS Trepte P, Kruse S, Kostova S, Hoffmann S, Buntru A, Tempelmeier A, Secker C, Diez L, Schulz A, Klockmeier K, Zenkner M, Golusik S, Rau K, Schnoegl S, Garner CC, Wanker EE. TITLE LuTHy: a double‐readout bioluminescence‐based two‐hybrid technology for quantitative mapping of protein–protein interactions in mammalian cells JOURNAL Molecular Systems Biology (2018) 14, e8071 REFERENCE 2 (bases 1 to 5967) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5967 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 907..936 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 937..1449 /codon_start=1 /product="NanoLuc(R) luciferase" /label=Nluc /note="human codon-optimized" /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR LCERILA" primer_bind complement(1561..1578) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1567..1791 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1837..2265 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1924..1943) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2134..2155 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2274..2294) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2279..2608 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2459..2594 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2521..2540 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2675..3469 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2729..2748) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3339..3358 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3643..3764 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3680..3699) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3734..3753 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3813..3829) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3813..3829) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3826..3848) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3837..3853 /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)." promoter complement(3861..3891) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3906..3927 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4044..4061) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4215..4800) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4295..4314) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4971..5831) /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 5594..5613 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5832..5936) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 accatggaac agaaactgat ctctgaagaa gacctggtct tcacactcga agatttcgtt 961 ggggactggc gacagacagc cggctacaac ctggaccaag tccttgaaca gggaggtgtg 1021 tccagtttgt ttcagaatct cggggtgtcc gtaactccga tccaaaggat tgtcctgagc 1081 ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc cgtatgaagg tctgagcggc 1141 gaccaaatgg gccagatcga aaaaattttt aaggtggtgt accctgtgga tgatcatcac 1201 tttaaggtga tcctgcacta tggcacactg gtaatcgacg gggttacgcc gaacatgatc 1261 gactatttcg gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta 1321 acagggaccc tgtggaacgg caacaaaatt atcgacgagc gcctgatcaa ccccgacggc 1381 tccctgctgt tccgagtaac catcaacgga gtgaccggct ggcggctgtg cgaacgcatt 1441 ctggcgtgaa agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattctgca 1501 gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag 1561 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 1621 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 1681 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 1741 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 1801 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 1861 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 1921 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 1981 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 2041 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 2101 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 2161 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 2221 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 2281 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 2341 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 2401 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 2461 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 2521 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 2581 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 2641 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 2701 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 2761 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 2821 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 2881 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 2941 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 3001 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 3061 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 3121 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 3181 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 3241 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 3301 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 3361 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 3421 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 3481 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 3541 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 3601 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 3661 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 3721 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 3781 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 3841 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 3901 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 3961 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 4021 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 4081 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 4141 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 4201 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 4261 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 4321 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4381 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 4441 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4501 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4561 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4621 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 4681 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4741 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 4801 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 4861 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 4921 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 4981 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 5041 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 5101 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 5161 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 5221 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 5281 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 5341 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 5401 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 5461 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 5521 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 5581 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 5641 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 5701 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 5761 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 5821 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 5881 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 5941 tttccccgaa aagtgccacc tgacgtc //