LOCUS Exported 8630 bp ds-DNA circular SYN 12-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(-8a) 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 12, 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 293..596 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 597..800 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 751..771 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 797..821 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind 926..942 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 926..942 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 953..971 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 953..970 /label=SP6 /note="SP6 promoter, forward primer" protein_bind 972..996 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(4607..4628) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4868..4887) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5215..5236 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(5326..5350) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(5351..5370) /label=T7 /note="T7 promoter, forward primer" promoter complement(5352..5370) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5381..5398) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5381..5397) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5390..5412) /label=M13/pUC Forward /note="In lacZ gene" polyA_signal 5802..5936 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5839..5858) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5893..5912 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6062..6516 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6149..6168) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6358..6379 /label=F1ori-F /note="F1 origin, forward primer" CDS 6803..7663 /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(7021..7040) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" protein_bind complement(7765..7798) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." rep_origin 7876..8464 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8365..8384 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8618..5 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 61 attaatgcag agcttgcaat tcgcgctttt tcaatattat tgaagcattt atcagggtta 121 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 181 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 241 aacctataaa aataggcgta ttacgaggcc ctttcacgca ttagatgcat gtcgttacat 301 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 361 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 421 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 481 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 541 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga 601 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa 661 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc 721 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg 781 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac 841 gctgttttga cctccataga agacaccggg accgatccag cctccggact ctagcctagg 901 ccgcgggacg gataacaatt tcacacagga aacagctatg accattaggc ctatttaggt 961 gacactatag aacaagtttg tacaaaaaag caggctggta ccggtccgga attcccggga 1021 tatcgtcgac ccacgcgtcc gttaagttcc gcgcgcaggc caggctgcaa cctgacggcc 1081 agatccctcg ctgtcctagt cgctgctcct tggagtcatg ttcccagccg ccccttctcc 1141 gcggaccccg ggtaccgggt cccgaagggg cccgctggcc ggactcgggc ccggctccac 1201 gccccggacg gctagcagga agggtctgcc cctggggtct gcagtcagct ccccagtgct 1261 cttctcgccg gtcggccggc gtagctcgct aagctcgcgg ggaacaccaa cacgaatgtt 1321 cccacaccac tccataactg agtctgtgaa ctatgatgtg aaaacgtttg gatcttctct 1381 tcctgttaaa gtcatggaag ccctaacatt ggctgaagtc gatgaccagc tgaccattaa 1441 catagatgaa ggtggatggg cttgtctggt gtgcaaagag aagctcatta tttggaagat 1501 tgctctgtca cctattacta agttatccgt ttgcaaagaa cttcagctgc cacctagtga 1561 tttccactgg agtgccgact tagtggctct ttcttactct tctccctcag gtgaagcaca 1621 ttctactcag gctgttgctg tcatggttgc caccagagaa ggatctatcc gctattggcc 1681 aagccttgct ggtgaagata cctacacaga ggcttttgta gattcgggag gtgataagac 1741 ttacagtttc ctaacagcag tgcagggagg aagttttatt ttgtcttcat caggaagcca 1801 actaattcgg ttgatacctg agagctcagg aaagattcat cagcatatcc tgcctcaggg 1861 gcaaggcatg ctttcaggaa ttggtcgaaa agtttcttct ctttttggaa ttttatctcc 1921 tagtagtgat ctcacacttt caagtgttct ctgggataga gagagatcaa gcttttatag 1981 cctgacgagt tcaaacatca gtaaatggga attagatgat tcttcagaaa agcatgcata 2041 cagttgggat ataaatagag ccctgaagga aaacattacc gatgctattt ggggatctga 2101 aagtaactat gaagctatta aagaaggagt caacattcga tatttggact ttaagcaaaa 2161 ctgtgatggg ctggtgattt tggcagcagc atggcactca gcagacaatc catgtctcat 2221 ctattactct ctgataacaa tagaagataa tggttgccaa atgtcagatg cagttactgt 2281 agaagtcact caatataatc caccttttca gtctgaagac ctgattttgt gtcagttgac 2341 ggtcccaaac ttttcaaacc agactgccta tctgtataac gaaagtgctg tctatgtgtg 2401 ctccacagga actgggaaat tttctcttcc ccaggagaaa attgtcttta atgcacaagg 2461 agatagtgtt ttaggtgctg gtgcctgtgg tggtgttcct atcatttttt ctagaaacag 2521 tggactggtg tctattactt caagggaaaa tgtgtctata ttggcagaag acttggaagg 2581 gtctttagca tcttcagttg ctggaccaaa cagtgagagt atgatttttg agaccactac 2641 aaagaatgaa actatagccc aggaagataa aatcaagttg ctgaaagctg cctttctgca 2701 atactgcaga aaagatttag gtcatgctca aatggtggtt gatgagctct tttcctctca 2761 ctctgatttg gattctgatt ctgaactaga cagggcagtt acccaaatca gtgtagacct 2821 gatggatgac tacccagcat ctgacccacg gtgggctgag tctgtccctg aggaagcacc 2881 tgggttcagc aatacgtcac tgattatcct tcaccagcta gaagacaaga tgaaagctca 2941 ctcttttctt atggacttta ttcatcaagt tggcttattt ggacgtctag gcagttttcc 3001 agttagaggg acaccgatgg ccactcgact gttgctctgt gagcatgccg aaaagctgtc 3061 agccgccatt gttctcaaga accaccactc ccggctttct gaccttgtca acacagccat 3121 attgattgct ttgaacaaga gggagtatga aatcccatcc aacctgactc ctgcagatgt 3181 ctttttcagg gaggtatccc aagtagatac catctgtgag tgcttactgg agcatgagga 3241 gcaagtcttg agggatgcac ctatggattc cattgaatgg gctgaagtgg tgatcaatgt 3301 gaacaatatt ctcaaggata tgctgcaggc tgctagtcat tatcgccaaa atagaaactc 3361 tttgtataga agagaagaat cactagaaaa agaacctgaa tatgttccat ggacggcaac 3421 aagtggtcct ggtggcatcc gaacggtaat aatacgccag catgagattg tcctgaaggt 3481 ggcttatcca caggcagaca gcaacctccg aaacatcgtg accgagcagc tggtagccct 3541 gatcgattgc ttcctggatg gttatgtttc tcagcttaag tctgtggata aatccagtaa 3601 tcgggaaaga tatgacaatc tggagatgga atacctacag aaaagatcag atctcttatc 3661 tcctcttctt tcactaggcc agtacctgtg ggctgcttct ctagcagaga aatactgtga 3721 ctttgatata ttggtacaaa tgtgtgagca gactgacaac cagagccgac tccagcgcta 3781 catgacccag tttgctgatc agaatttttc agactttctc ttccgttggt atctggagaa 3841 aggaaagcga ggcaaattat tatctcagcc catttctcag catggacagt tggcaaattt 3901 tttgcaagct catgaacatc tcagctggtt acatgaaatt aatagccaag aattagaaaa 3961 ggctcatgca acacttctgg gtttggcaaa tatggaaact cgttactttg caaagaagaa 4021 aacccttctt ggcttgagta aattggctgc attagcttca gacttttcag aggatatgct 4081 acaagaaaaa attgaagaaa tggctgagca ggagcgcttt ctactgcatc aggagaccct 4141 acctgaacag ctgctggcgg agaaacagct aaatctcagt gcgatgccag tattgactgc 4201 accacaactc attggtctat atatctgtga agaaaataga agagctaatg aatatgattt 4261 caagaaagct ttggacttgt tggaatatat tgatgaggaa gaagatataa atataaatga 4321 tctaaaactg gaaatccttt gcaaagctct tcagagagat aactggtcca gttctgatgg 4381 caaagatgat ccaattgaag tatctaaaga cagtatattt gtgaagatct tacagaaact 4441 tttaaaagat ggcattcagc tcagtgagta cttaccggag gtgaaagacc tgctacaagc 4501 ggatcagctt ggaagcttaa agtccaatcc ttacttcgag tttgttttga aagcaaatta 4561 tgaatattat gttcagggag agctgttcac cggggtggtg cccatcctgg tcgagctgga 4621 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta 4681 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac 4741 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa 4801 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt 4861 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct 4921 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca 4981 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa 5041 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc 5101 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca 5161 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt 5221 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta 5281 agcggccgct ctagagtatc cctcgagggg cccaagctta cgcgtaccca gctttcttgt 5341 acaaagtggt ccctatagtg agtcgtatta taagctaggc actggccgtc gttttacaac 5401 gtcgtgactg ggaaaactgc tagcttggga tctttgtgaa ggaaccttac ttctgtggtg 5461 tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt 5521 taagtgtata atgtgttaaa ctagctgcat atgcttgctg cttgagagtt ttgcttactg 5581 agtatgattt atgaaaatat tatacacagg agctagtgat tctaattgtt tgtgtatttt 5641 agattcacag tcccaaggct catttcaggc ccctcagtcc tcacagtctg ttcatgatca 5701 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc 5761 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt 5821 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac 5881 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcgatc 5941 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg ctggcgtaat 6001 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg 6061 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 6121 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 6181 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 6241 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 6301 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg 6361 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 6421 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 6481 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt 6541 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgcc 6601 aggtcttgga ctggtgagaa cggcttgctc ggcagcttcg atgtgtgctg gagggagaat 6661 aaaggtctaa gatgtgcgat agagggaagt cgcattgaat tatgtgctgt gtagggatcg 6721 ctggtatcaa atatgtgtgc ccacccctgg catgagacaa taaccctgat aaatgcttca 6781 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 6841 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 6901 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 6961 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct 7021 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat 7081 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 7141 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 7201 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 7261 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 7321 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 7381 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 7441 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 7501 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc 7561 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 7621 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 7681 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 7741 gatccttttt gataatctca tgccataact tcgtataatg tatgctatac gaagttatgg 7801 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 7861 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 7921 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 7981 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 8041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 8101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 8161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 8221 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 8281 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 8341 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 8401 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 8461 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 8521 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 8581 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga //