LOCUS Exported 7376 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Flag-HA-BRCC3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7376) AUTHORS Sowa ME, Bennett EJ, Gygi SP, Harper JW TITLE Defining the human deubiquitinating enzyme interaction landscape. JOURNAL Cell. 2009 Jul 23. 138(2):389-403. PUBMED 19615732 REFERENCE 2 (bases 1 to 7376) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7376 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 107..128 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /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)." LTR 359..873 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" CDS complement(949..1548) /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(1032..1052) /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind 1529..1548 /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" misc_feature 1560..2112 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1722..1741) /label=IRES-F /note="IRES internal ribosome entry site, forward primer" primer_bind 1951..1968 /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" protein_bind 2142..2166 /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(3115..3139) /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS complement(3155..3181) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS complement(3194..3217) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" regulatory 3217..3226 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(3239..3655) /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind complement(3255..3271) /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(3287..3309) /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" misc_feature 3722..4063 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" LTR 4127..4643 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" primer_bind complement(4843..4862) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(4990..5012) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(5206..5225) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5325..5347 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5385..5403) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5471..5575 /gene="bla" /label=AmpR promoter CDS 5576..6436 /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(5794..5813) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6607..7195 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7096..7115 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7349..7366 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacaattctc cccagctctt tctgcgctgt ccataatgaa 241 ctatttcagg agtattggag tatgtggatt caaaacagtg gctaattgag tcaggattac 301 ttaatattcg actcctaccc tcagaatgtt gttctccaac ttcaagaaac tgttacccat 361 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga 421 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgacg 481 cagtctatcg gaggactggc gcgccgagtg aggggttgtg ggctctttta ttgagctcgg 541 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca 601 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc 661 tgagggcggg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg 721 cttaccacag atatcctgtt tggcccatat tctgctgtct ctctgttcct aaccttgatc 781 tgaacttctc tattctcagt tatgtatttt ccatgccttg caaaatggcg ttacttaagc 841 tagcttgcca aacctacagg tggggtcttt cattcccccc tttttctgga gactaaataa 901 aatcttttat tttatcgata agcttggctg caggtcgaca tcgatgcatc aggcaccggg 961 cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt 1021 gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg 1081 caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt 1141 gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg 1201 gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc 1261 ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt 1321 ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac 1381 gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac 1441 ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg 1501 gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg ttgtggccat 1561 attatcatcg tgtttttcaa aggaaaacca cgtccccgtg gttcgggggg cctagacgtt 1621 tttttaacct cgactaaaca catgtaaagc atgtgcaccg aggccccaga tcagatccca 1681 tacaatgggg taccttctgg gcatccttca gccccttgtt gaatacgctt gaggagagcc 1741 atttgactct ttccacaact atccaactca caacgtggca ctggggttgt gccgcctttg 1801 caggtgtatc ttatacacgt ggcttttggc cgcagaggca cctgtcgcca ggtggggggt 1861 tccgctgcct gcaaagggtc gctacagacg ttgtttgtct tcaagaagct tccagaggaa 1921 ctgcttcctt cacgacattc aacagacctt gcattccttt ggcgagaggg gaaagacccc 1981 taggaatgct cgtcaagaag acagggccag gtttccgggc cctcacattg ccaaaagacg 2041 gcaatatggt ggaaaataac atatagacaa acgcacaccg gccttattcc aagcggcttc 2101 ggccagtaac gttagggggg gggggcggaa ttcgttatcg aaccactttg tacaagaaag 2161 ttgggtattc tagagaagaa agttcttgca taagctcttc cttttcttgt tgtaattcct 2221 gcaaatgctg ttggttttgc tccagtctgt cctccaacca ctgtaggaga ggcccgctga 2281 ctgccgacat ctgactgcac agattcttgg taaacactga gccattatgg atcttggtta 2341 ctgagtccag atgtgtaagg ctgtggatcc tcctatacgc atcctgctcc tcctggcaca 2401 ggatcttggg cagctctact gctgattcaa ggcacacttt cccgatagtg acatgaggta 2461 caatatggat tgggatttcg attctctcat acctttcctc ttccttctgg ccctcaatgg 2521 agatgtgctg gctggagctc cagaagtctc gtggaccatg aagggactct gaactctttt 2581 gggcctgtat ggattggaag caagtgtaga gtacccggcc agtctttgtg ttcttatctt 2641 ctatgaaaca ggaaaaaata agtcctacaa agccttgatc catcatctgg tacatggctt 2701 gtgtgcgaac atcaacatgt gaaggccaaa cagttatatg aggatgggaa tgataccagc 2761 ccacaactct catggggcgg cctgtcagtt cagccaacct ctctgcctct gttgaagctg 2821 cagacagctg ctctggagaa atttctactc ggtccttcct cttatcagaa cgtcgtaaga 2881 tgatgacaga atgaatgtga acaattctga cggcatcaac cttttcagca actgtgcgca 2941 tttcagttcc agtatatgca aatttggagt cactccttgt atcatcgttc aactccccta 3001 tgcacagccc cattacttcc tccttctctg tgctcagagc gtggttgaga caaacgagga 3061 aagcgtcaga ctcgagatga accgcctgca ccgcctgcac cacctgcacc gccatgccaa 3121 cttttttgta caaacttgtt tgataacctc gagcagcgta atctggaaca tcgtatgggt 3181 atcctccatc gagcttgtca tcgtcatcct tgtagtccat ggtggcagat ctaattccgg 3241 cgcctagaga aggagtgagg gctggataaa gggaggatcg aggcggggtc gaacgaggag 3301 gttcaagggg gagagacggg gcggatggag gaagaggagg cggaggctta gggtgtacaa 3361 agggcttgac ccagggaggg gggtcaaaag ccaaggcttc ccaggtcacg atgtagggga 3421 cctggtctgg gtgtccatgc gggccaggtg aaaagacctt gatcttaacc tgggtgatga 3481 ggtctcggtt aaaggtgccg tctcgcggcc atccgacgtt aaaggttggc cattctgcag 3541 agcagaaggt aacccaacgt ctcttcttga catctaccga ctggttgtga gcgatccgct 3601 cgacatcttt ccagtgacct aaggtcaaac ttaagggagt ggtaacagtc tggccctaat 3661 tttcagacaa atacagaaac acagtcagac agagacaaca cagaacgatg ctgcagcgct 3721 gcagcagaca agacgcgcgg cttcggttcc aaaccgaaag caaaaattca gacggaggcg 3781 ggaactgttt taggttctcg tctcctacca gaaccacata tcctgacggg gtcggattcc 3841 acatcgactc ccttcctcag gtcgggccac aaaaacggcc cccaaagtcc ctgggacgtc 3901 tcccagggtt gcggccgggt gttcagaact cgtcagttcc accacgggtc cgccagatac 3961 agagctagtt agctaactag tacagacgca ggcgcaaaca ttagatgccg gcacaaacac 4021 gcacaaagac agagacagac acgaaacgac cgctggccag cttacctccc ggcggggggg 4081 tcggtggtcc ctgggcaggg gtctccaaat ctcggtggaa cctccaaatg aaagaccccc 4141 gaggtgggca gtcaatcaat ctgaggagac cctcccaagg atcagcgagt ccacgattcg 4201 gatgcaaaca gcaagaggct ttattgggaa tacgggtacc cgggcgacgc agtctatcgg 4261 aggactggcg cgccgagtga ggggttgtgg gctcttttat tgagctcggg gagcagaagc 4321 gcgcgaacag aagcgagaag cgaactgatt ggttagttca aataaggcac agggtcattt 4381 caggtccttg gggcaccctg gaaacatctg atggttctct agaaactgct gagggcggga 4441 ccgcatctgg ggaccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga 4501 tatcctgttt ggcccatatt ctgctgtctc tctgttccta accttgatct gaacttctct 4561 attctcagtt atgtattttc catgccttgc aaaatggcgt tacttaagct agcttgccaa 4621 acctacaggt ggggtctttc attcccccct ttttctggag actaaataaa atcttttatt 4681 ttatctatgg ctcgtactct ataggcttca gctggtgata ttgttgagtc aaaactagag 4741 cctggaccac tgatatcctg tctttaacaa attggactaa tcgcctctac gccggacgca 4801 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca 4861 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta 4921 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc 4981 ttgcgccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg 5041 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg 5101 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct 5161 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga 5221 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc 5281 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg 5341 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct 5401 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg 5461 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc 5521 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag 5581 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt 5641 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt 5701 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga 5761 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat 5821 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga 5881 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag 5941 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg 6001 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg 6061 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt 6121 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg 6181 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc 6241 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg 6301 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac 6361 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact 6421 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa 6481 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa 6541 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg 6601 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc 6661 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac 6721 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca 6781 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt 6841 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc 6901 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg 6961 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc 7021 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac 7081 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct 7141 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc 7201 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt 7261 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac 7321 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaag //