LOCUS Exported 5681 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses HA SUMO2 Wild Type in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA3 HA-Sumo2 WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5681) AUTHORS Bekes M, Prudden J, Srikumar T, Raught B, Boddy MN, Salvesen GS TITLE The dynamics and mechanism of SUMO chain deconjugation by SUMO-specific proteases. JOURNAL J Biol Chem. 2011 Mar 25;286(12):10238-47. Epub 2011 Jan 19. PUBMED 21247896 REFERENCE 2 (bases 1 to 5681) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5681 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(51..71) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 56..385 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 236..371 /label=SV40 ori /note="SV40 origin of replication" primer_bind 298..317 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 452..1246 /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(506..525) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1116..1135 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 1420..1541 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1457..1476) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1511..1530 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(1590..1606) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1590..1606) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1603..1625) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1614..1630 /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(1638..1668) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1683..1704 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1821..1838) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1992..2580) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2072..2091) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2751..3611) /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 3374..3393 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3612..3716) /gene="bla" /label=AmpR promoter primer_bind complement(3791..3810) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 3982..4361 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4362..4565 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4516..4536 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4610..4629 /label=T7 /note="T7 promoter, forward primer" promoter 4610..4628 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 4651..4677 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 4675..4950 /codon_start=1 /product="human small ubiquitin-related modifier 2" /label=SUMO2 /translation="ADEKPKEGVKTENNDHINLKVAGQDGSVVQFKIKRHTPLSKLMKA YCERQGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQQTGG" promoter complement(4981..4999) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4982..4999) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(5019..5036) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5025..5249 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5295..42 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5382..5401) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5592..5613 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg 61 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 121 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 181 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 241 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 301 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt 361 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc 421 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 481 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 541 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 601 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct 661 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 721 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 781 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 841 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 901 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 961 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 1021 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 1081 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 1141 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 1201 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 1261 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 1321 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 1381 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 1441 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 1501 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 1561 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 1621 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 1681 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 1741 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 1801 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 1861 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 1921 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 1981 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 2041 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 2101 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 2161 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 2221 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 2281 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 2341 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 2401 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 2461 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 2521 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 2581 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 2641 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 2701 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 2761 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 2821 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 2881 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 2941 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 3001 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 3061 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 3121 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 3181 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 3241 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 3301 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 3361 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 3421 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 3481 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 3541 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 3601 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 3661 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 3721 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt 3781 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 3841 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 3901 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 3961 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 4021 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 4081 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 4141 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 4201 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 4261 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 4321 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 4381 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 4441 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 4501 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 4561 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 4621 actataggga gacccaagct tatggatgcc tacccatacg acgtaccaga ctacgccgac 4681 gaaaagccca aggaaggagt caagactgag aacaacgatc atattaattt gaaggtggcg 4741 gggcaggatg gttctgtggt gcagtttaag attaagaggc atacaccact tagtaaacta 4801 atgaaagcct attgtgaacg acagggattg tcaatgaggc agatcagatt ccgatttgac 4861 gggcaaccaa tcaatgaaac agacacacct gcacagttgg aaatggagga tgaagataca 4921 attgatgtgt tccaacagca gacgggaggt tgactcgagc atgcatctag agggccctat 4981 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg 5041 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 5101 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 5161 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 5221 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc 5281 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 5341 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 5401 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 5461 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 5521 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 5581 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 5641 ttcttttgat ttataaggga ttttgccgat ttcggcctat t //