LOCUS HA-Ubiquitin 5671 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION Mammalian expression of human ubiquitin with HA tag. ACCESSION . VERSION . KEYWORDS HA-Ubiquitin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5671) AUTHORS Kamitani T, Kito K, Nguyen HP, Yeh ET TITLE Characterization of NEDD8, a developmentally down-regulated ubiquitin-like protein. JOURNAL J Biol Chem. 1997 Nov 7. 272(45):28557-62. PUBMED 9353319 COMMENT This file is created by Vector NTI //gt12365.cn/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|gt12365.cn| COMMENT VNTNAME|HA-Ubiquitin| FEATURES Location/Qualifiers source 1..5671 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(13..33) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 18..347 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 198..333 /label=SV40 ori /note="SV40 origin of replication" primer_bind 260..279 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 414..1208 /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(468..487) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1078..1097 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 1382..1503 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1419..1438) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1473..1492 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(1552..1568) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1552..1568) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1565..1587) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1576..1592 /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(1600..1630) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1645..1666 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1783..1800) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1954..2542) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2034..2053) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2713..3573) /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 3336..3355 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3574..3678) /gene="bla" /label=AmpR promoter primer_bind complement(3753..3772) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 3944..4323 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4324..4527 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4478..4498 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4572..4591 /label=T7 /note="T7 promoter, forward primer" promoter 4572..4590 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 4607..4633 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 4643..4870 /codon_start=1 /gene="Homo sapiens UBB" /product="human ubiquitin B" /label=ubiquitin /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG" promoter complement(4933..4951) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4934..4951) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(4971..4988) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4977..5201 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5247..4 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5334..5353) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5544..5565 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag 61 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 361 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg 421 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 481 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 541 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg 601 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 661 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 721 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 781 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 841 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 901 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg 961 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 1021 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 1081 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 1141 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 1201 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc 1261 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg 1321 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc 1381 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 1441 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 1501 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct 1561 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 1621 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 1681 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 1741 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 1801 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 1861 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 1921 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 1981 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 2041 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 2101 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 2161 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 2221 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 2281 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 2341 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 2401 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 2461 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 2521 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 2581 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 2641 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 2701 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 2761 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 2821 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 2881 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 2941 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 3001 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 3061 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 3121 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 3181 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 3241 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 3301 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 3361 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 3421 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 3481 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 3541 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 3601 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 3661 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 3721 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 3781 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt 3841 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg 3901 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact 3961 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc 4021 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 4081 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 4141 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 4201 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 4261 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 4321 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 4381 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg 4441 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta 4501 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg 4561 cttatcgaaa ttaatacgac tcactatagg gagacccaag cttatgtacc catacgatgt 4621 tccggattac gctagcggat ccatgcagat cttcgtgaaa acccttaccg gcaagaccat 4681 cacccttgag gtggagccca gtgacaccat cgaaaatgtg aaggccaaga tccaggataa 4741 ggaaggcatt ccccccgacc agcagaggct catctttgca ggcaagcagc tggaagatgg 4801 ccgtactctt tctgactaca acatccagaa ggagtcgacc ctgcacctgg tcctgcgtct 4861 gagaggtggt tagaattctg cagatatcca tcacactggc ggccgctcga gcatgcatct 4921 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt 4981 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 5041 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 5101 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 5161 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 5221 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 5281 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 5341 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 5401 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 5461 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 5521 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 5581 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 5641 aaatgagctg atttaacaaa aatttaacgc g //