LOCUS Exported 7400 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Flag-HA-PSMD7 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7400) 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 7400) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7400 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1..417 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 347..369 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 385..401 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" regulatory 430..439 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 439..462 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 475..501 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" protein_bind 517..541 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" RBS 1489..1500 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" protein_bind complement(1514..1538) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 1568..2120 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1712..1729) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1939..1958 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2132..2731 /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(2132..2151) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2628..2648 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 2807..3321 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 3483..3499 /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(3507..3537) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3552..3573 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3690..3707) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3861..4449) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3941..3960) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4620..5480) /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 5243..5262 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5481..5585) /gene="bla" /label=AmpR promoter primer_bind 5653..5671 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5709..5731) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5831..5850 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6044..6066 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6194..6213 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 6413..6929 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 6993..7334 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" ORIGIN 1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg 61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca 121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc 181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc 241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta 301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct 361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga 421 attagatctg ccaccatgga ctacaaggat gacgatgaca agctcgatgg aggataccca 481 tacgatgttc cagattacgc tgctcgaggt tatcaaacaa gtttgtacaa aaaagttggc 541 atgccggagc tggcagtgca gaaggtggtg gtccaccccc tggtgctgct cagtgtggtg 601 gatcatttca accgaatcgg caaggttgga aaccagaagc gtgttgttgg tgtgcttttg 661 gggtcatggc aaaagaaagt acttgatgta tcgaacagtt ttgcagttcc ttttgatgaa 721 gatgacaaag acgattctgt atggttttta gaccatgatt atttggaaaa catgtatgga 781 atgtttaaga aagtcaatgc cagggaaaga atagttggct ggtaccacac aggccctaaa 841 ctacacaaga atgacattgc catcaacgaa ctcatgaaaa gatactgtcc taattccgta 901 ttggtcatca ttgatgtgaa gccgaaggac ctagggctgc ctacagaagc gtacatttca 961 gtggaagaag tccatgatga tggaactcca acctcgaaaa catttgaaca cgtgaccagt 1021 gaaattggag cagaggaagc tgaggaagtt ggagttgaac acttgttacg agatatcaaa 1081 gacacgacgg tgggcactct gtcccagcgg atcacaaacc aggtccatgg tttgaaggga 1141 ctgaactcca agcttctgga tatcaggagc tacctggaaa aagtcgccac aggcaagctg 1201 cccatcaacc accagatcat ctaccagctg caggacgtct tcaacctgct gccagatgtc 1261 agcctgcagg agttcgtcaa ggccttttac ctgaagacca atgaccagat ggtggtagtg 1321 tacttggcct cgctgatccg ttccgtggtc gccctgcaca acctcatcaa caacaagatt 1381 gccaaccggg atgcagagaa gaaagaaggg caggagaaag aagagagcaa aaaggatagg 1441 aaagaggaca aggagaaaga taaagataag gaaaagagtg atgtaaagaa agaggagaaa 1501 aaggagaaaa agtaaccaac tttcttgtac aaagtggttc gataacgaat tccgcccccc 1561 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg 1621 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc 1681 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg 1741 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg 1801 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca 1861 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata 1921 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc 1981 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt 2041 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt 2101 tgaaaaacac gatgataata tggccacaac catgaccgag tacaagccca cggtgcgcct 2161 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta 2221 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca 2281 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg 2341 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga 2401 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga 2461 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt 2521 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc 2581 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt 2641 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac 2701 ctggtgcatg acccgcaagc ccggtgcctg atgcatcgat gtcgacctgc agccaagctt 2761 atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac 2821 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaatacataa 2881 ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa tatgggccaa 2941 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc 3001 agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca 3061 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct 3121 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg 3181 ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac cctcttgcag 3241 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac 3301 ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca acattctgag 3361 ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga atccacatac 3421 tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg agaattgtga 3481 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 3541 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 3601 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 3661 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 3721 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 3781 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 3841 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 3901 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 3961 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 4021 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 4081 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 4141 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 4201 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 4261 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 4321 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 4381 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 4441 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 4501 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 4561 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 4621 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 4681 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 4741 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 4801 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 4861 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 4921 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 4981 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 5041 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 5101 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 5161 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 5221 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 5281 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 5341 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 5401 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 5461 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 5521 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 5581 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 5641 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 5701 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 5761 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg 5821 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata 5881 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc 5941 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg 6001 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt 6061 aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca 6121 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc 6181 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg 6241 tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga 6301 tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt 6361 acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag 6421 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa 6481 tacataactg agaatagaga agttcagatc aaggttagga acagagagac agcagaatat 6541 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat 6601 ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg 6661 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct 6721 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact 6781 cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct 6841 cttgctgttt gcatccgaat cgtggactcg ctgatccttg ggagggtctc ctcagattga 6901 ttgactgccc acctcggggg tctttcattt ggaggttcca ccgagatttg gagacccctg 6961 cccagggacc accgaccccc ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg 7021 tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt 7081 tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg 7141 gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag 7201 gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga 7261 acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg 7321 cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc 7381 tgtatttgtc tgaaaattag //