LOCUS Exported 8630 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Mammalian expression of Nup133 fused at its carboxyl-terminus to mEGFP with no linker and a total net fusion of (-8 amino acids) . ACCESSION . VERSION . KEYWORDS Nup133_mEGFP(-8b) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8630) TITLE Simon lab plasmids REFERENCE 2 (bases 1 to 8630) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8630 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 181..484 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 485..688 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 639..659 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 685..709 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind 814..830 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 814..830 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 841..859 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 841..858 /label=SP6 /note="SP6 promoter, forward primer" protein_bind 860..884 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(4495..4516) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4756..4775) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5103..5124 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(5214..5238) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(5239..5258) /label=T7 /note="T7 promoter, forward primer" promoter complement(5240..5258) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5269..5286) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5269..5285) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5278..5300) /label=M13/pUC Forward /note="In lacZ gene" polyA_signal 5690..5824 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5727..5746) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5781..5800 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5950..6404 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6037..6056) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6246..6267 /label=F1ori-F /note="F1 origin, forward primer" CDS 6691..7551 /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(6909..6928) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" protein_bind complement(7653..7686) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." rep_origin 7764..8352 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8253..8272 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8506..8523 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 61 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc 121 atgacattaa cctataaaaa taggcgtatt acgaggccct ttcacgcatt agatgcatgt 181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 661 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg 721 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct 781 agcctaggcc gcgggacgga taacaatttc acacaggaaa cagctatgac cattaggcct 841 atttaggtga cactatagaa caagtttgta caaaaaagca ggctggtacc ggtccggaat 901 tcccgggata tcgtcgaccc acgcgtccgt taagttccgc gcgcaggcca ggctgcaacc 961 tgacggccag atccctcgct gtcctagtcg ctgctccttg gagtcatgtt cccagccgcc 1021 ccttctccgc ggaccccggg taccgggtcc cgaaggggcc cgctggccgg actcgggccc 1081 ggctccacgc cccggacggc tagcaggaag ggtctgcccc tggggtctgc agtcagctcc 1141 ccagtgctct tctcgccggt cggccggcgt agctcgctaa gctcgcgggg aacaccaaca 1201 cgaatgttcc cacaccactc cataactgag tctgtgaact atgatgtgaa aacgtttgga 1261 tcttctcttc ctgttaaagt catggaagcc ctaacattgg ctgaagtcga tgaccagctg 1321 accattaaca tagatgaagg tggatgggct tgtctggtgt gcaaagagaa gctcattatt 1381 tggaagattg ctctgtcacc tattactaag ttatccgttt gcaaagaact tcagctgcca 1441 cctagtgatt tccactggag tgccgactta gtggctcttt cttactcttc tccctcaggt 1501 gaagcacatt ctactcaggc tgttgctgtc atggttgcca ccagagaagg atctatccgc 1561 tattggccaa gccttgctgg tgaagatacc tacacagagg cttttgtaga ttcgggaggt 1621 gataagactt acagtttcct aacagcagtg cagggaggaa gttttatttt gtcttcatca 1681 ggaagccaac taattcggtt gatacctgag agctcaggaa agattcatca gcatatcctg 1741 cctcaggggc aaggcatgct ttcaggaatt ggtcgaaaag tttcttctct ttttggaatt 1801 ttatctccta gtagtgatct cacactttca agtgttctct gggatagaga gagatcaagc 1861 ttttatagcc tgacgagttc aaacatcagt aaatgggaat tagatgattc ttcagaaaag 1921 catgcataca gttgggatat aaatagagcc ctgaaggaaa acattaccga tgctatttgg 1981 ggatctgaaa gtaactatga agctattaaa gaaggagtca acattcgata tttggacttt 2041 aagcaaaact gtgatgggct ggtgattttg gcagcagcat ggcactcagc agacaatcca 2101 tgtctcatct attactctct gataacaata gaagataatg gttgccaaat gtcagatgca 2161 gttactgtag aagtcactca atataatcca ccttttcagt ctgaagacct gattttgtgt 2221 cagttgacgg tcccaaactt ttcaaaccag actgcctatc tgtataacga aagtgctgtc 2281 tatgtgtgct ccacaggaac tgggaaattt tctcttcccc aggagaaaat tgtctttaat 2341 gcacaaggag atagtgtttt aggtgctggt gcctgtggtg gtgttcctat cattttttct 2401 agaaacagtg gactggtgtc tattacttca agggaaaatg tgtctatatt ggcagaagac 2461 ttggaagggt ctttagcatc ttcagttgct ggaccaaaca gtgagagtat gatttttgag 2521 accactacaa agaatgaaac tatagcccag gaagataaaa tcaagttgct gaaagctgcc 2581 tttctgcaat actgcagaaa agatttaggt catgctcaaa tggtggttga tgagctcttt 2641 tcctctcact ctgatttgga ttctgattct gaactagaca gggcagttac ccaaatcagt 2701 gtagacctga tggatgacta cccagcatct gacccacggt gggctgagtc tgtccctgag 2761 gaagcacctg ggttcagcaa tacgtcactg attatccttc accagctaga agacaagatg 2821 aaagctcact cttttcttat ggactttatt catcaagttg gcttatttgg acgtctaggc 2881 agttttccag ttagagggac accgatggcc actcgactgt tgctctgtga gcatgccgaa 2941 aagctgtcag ccgccattgt tctcaagaac caccactccc ggctttctga ccttgtcaac 3001 acagccatat tgattgcttt gaacaagagg gagtatgaaa tcccatccaa cctgactcct 3061 gcagatgtct ttttcaggga ggtatcccaa gtagatacca tctgtgagtg cttactggag 3121 catgaggagc aagtcttgag ggatgcacct atggattcca ttgaatgggc tgaagtggtg 3181 atcaatgtga acaatattct caaggatatg ctgcaggctg ctagtcatta tcgccaaaat 3241 agaaactctt tgtatagaag agaagaatca ctagaaaaag aacctgaata tgttccatgg 3301 acggcaacaa gtggtcctgg tggcatccga acggtaataa tacgccagca tgagattgtc 3361 ctgaaggtgg cttatccaca ggcagacagc aacctccgaa acatcgtgac cgagcagctg 3421 gtagccctga tcgattgctt cctggatggt tatgtttctc agcttaagtc tgtggataaa 3481 tccagtaatc gggaaagata tgacaatctg gagatggaat acctacagaa aagatcagat 3541 ctcttatctc ctcttctttc actaggccag tacctgtggg ctgcttctct agcagagaaa 3601 tactgtgact ttgatatatt ggtacaaatg tgtgagcaga ctgacaacca gagccgactc 3661 cagcgctaca tgacccagtt tgctgatcag aatttttcag actttctctt ccgttggtat 3721 ctggagaaag gaaagcgagg caaattatta tctcagccca tttctcagca tggacagttg 3781 gcaaattttt tgcaagctca tgaacatctc agctggttac atgaaattaa tagccaagaa 3841 ttagaaaagg ctcatgcaac acttctgggt ttggcaaata tggaaactcg ttactttgca 3901 aagaagaaaa cccttcttgg cttgagtaaa ttggctgcat tagcttcaga cttttcagag 3961 gatatgctac aagaaaaaat tgaagaaatg gctgagcagg agcgctttct actgcatcag 4021 gagaccctac ctgaacagct gctggcggag aaacagctaa atctcagtgc gatgccagta 4081 ttgactgcac cacaactcat tggtctatat atctgtgaag aaaatagaag agctaatgaa 4141 tatgatttca agaaagcttt ggacttgttg gaatatattg atgaggaaga agatataaat 4201 ataaatgatc taaaactgga aatcctttgc aaagctcttc agagagataa ctggtccagt 4261 tctgatggca aagatgatcc aattgaagta tctaaagaca gtatatttgt gaagatctta 4321 cagaaacttt taaaagatgg cattcagctc agtgagtact taccggaggt gaaagacctg 4381 ctacaagcgg atcagcttgg aagcttaaag tccaatcctt acttcgagtt tgttttgaaa 4441 gcaaattatg aatattatgt tcaggaggag ctgttcaccg gggtggtgcc catcctggtc 4501 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat 4561 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc 4621 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac 4681 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc 4741 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc 4801 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc 4861 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag 4921 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg 4981 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc 5041 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat 5101 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg 5161 tacaagtaag cggccgctct agagtatccc tcgaggggcc caagcttacg cgtacccagc 5221 tttcttgtac aaagtggtcc ctatagtgag tcgtattata agctaggcac tggccgtcgt 5281 tttacaacgt cgtgactggg aaaactgcta gcttgggatc tttgtgaagg aaccttactt 5341 ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat 5401 aaaattttta agtgtataat gtgttaaact agctgcatat gcttgctgct tgagagtttt 5461 gcttactgag tatgatttat gaaaatatta tacacaggag ctagtgattc taattgtttg 5521 tgtattttag attcacagtc ccaaggctca tttcaggccc ctcagtcctc acagtctgtt 5581 catgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc 5641 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat 5701 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 5761 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 5821 gatcgatcct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattggct 5881 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg 5941 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca 6001 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct 6061 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt 6121 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac 6181 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct 6241 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt 6301 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca 6361 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg 6421 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 6481 ctcatgccag gtcttggact ggtgagaacg gcttgctcgg cagcttcgat gtgtgctgga 6541 gggagaataa aggtctaaga tgtgcgatag agggaagtcg cattgaatta tgtgctgtgt 6601 agggatcgct ggtatcaaat atgtgtgccc acccctggca tgagacaata accctgataa 6661 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 6721 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 6781 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 6841 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 6901 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt 6961 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 7021 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 7081 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 7141 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 7201 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 7261 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 7321 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 7381 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 7441 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 7501 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 7561 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 7621 taggtgaaga tcctttttga taatctcatg ccataacttc gtataatgta tgctatacga 7681 agttatggca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 7741 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 7801 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 7861 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg 7921 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 7981 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 8041 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 8101 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 8161 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 8221 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 8281 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 8341 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 8401 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 8461 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc 8521 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat 8581 taatgcagag cttgcaattc gcgctttttc aatattattg aagcatttat //