LOCUS Exported 6155 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of ubiquitin with an N-terminal HA tag. ACCESSION . VERSION . KEYWORDS pCMV6-AN-HA_Ubiquitin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6155) AUTHORS Ashton NW, Valles GJ, Jaiswal N, Bezsonova I, Woodgate R TITLE DNA polymerase iota interacts with both the TRAF-like and UBL1-2 domains of USP7 JOURNAL Journal of Molecular Biology. 2020. REFERENCE 2 (bases 1 to 6155) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6155 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 140..162 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 154..171 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 155..171 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 332..711 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 712..915 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 866..886 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 941..960 /label=T7 /note="T7 promoter, forward primer" promoter 941..959 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1012..1038 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1057..1284 /codon_start=1 /product="human ubiquitin" /label=ubiquitin /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG" primer_bind complement(1349..1365) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" polyA_signal 1376..1998 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(1575..1594) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(2022..2042) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2027..2356 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2207..2342 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2269..2288 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2423..3217 /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(2477..2496) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3087..3106 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3391..3512 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3428..3447) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3482..3501 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3561..3577) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3574..3596) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3585..3601 /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(3609..3639) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3654..3675 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3792..3809) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3963..4551) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4043..4062) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4722..5582) /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 5345..5364 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5583..5687) /gene="bla" /label=AmpR promoter rep_origin 5714..14 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5801..5820) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6011..6032 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg 61 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt 121 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg 181 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa 241 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat 301 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag 361 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 421 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 481 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat 541 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct 601 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg 661 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg 721 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc 781 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa 841 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc 901 tatataagca gagctcgttt agtgaaccgt cagaattttg taatacgact cactataggg 961 cggccgggaa ttcgtcgact ggatccggta ccgaggagat ctgccaccat gtacccatac 1021 gatgttccag attacgctgc gatcgccgat atccatatgc agatttttgt gaaaaccctg 1081 accggcaaaa ccattaccct ggaagtggaa ccgagcgata ccattgaaaa cgtgaaagcg 1141 aaaattcagg ataaagaagg cattccgccg gatcagcagc gtctgatttt tgcgggtaaa 1201 cagctggaag atggtcgtac cctgagcgat tataacattc agaaagaaag caccctgcat 1261 ctggttctgc gtctgcgtgg tggttaagga tccaagcttc tcgagacgcg ttaagcggcc 1321 gcactcgagg tttaaacggc cggccgcggt catagctgtt tcctgaacag atcccgggtg 1381 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc 1441 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta 1501 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg 1561 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac 1621 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg 1681 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt 1741 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc 1801 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt 1861 aaaataacta taccagcagg aggacgtcca gacacagcat aggctacctg gccatgccca 1921 accggtggga catttgagtt gcttgcttgg cactgtcctc tcatgcgttg ggtccactca 1981 gtagatgcct gttgaattgg gtacgcggcc agcttggctg tggaatgtgt gtcagttagg 2041 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 2101 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 2161 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 2221 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 2281 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 2341 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga 2401 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 2461 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 2521 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 2581 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac 2641 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 2701 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 2761 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 2821 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 2881 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 2941 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 3001 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 3061 tgtggccgac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 3121 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 3181 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg 3241 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 3301 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 3361 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 3421 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 3481 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc 3541 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 3601 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 3661 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 3721 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 3781 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 3841 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 3901 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 3961 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 4021 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 4081 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 4141 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 4201 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 4261 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 4321 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 4381 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 4441 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 4501 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 4561 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 4621 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 4681 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 4741 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 4801 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 4861 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 4921 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 4981 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 5041 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 5101 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 5161 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 5221 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 5281 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 5341 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 5401 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 5461 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 5521 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 5581 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 5641 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 5701 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 5761 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 5821 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 5881 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 5941 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 6001 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 6061 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 6121 taacaaaaat ttaacgcgaa ttttaacaaa atatt //