LOCUS Exported 7674 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses mouse TRAF3IP3 in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA-mTRAF3ip3-CFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7674) TITLE Sun Lab TRAF3ip and Tbkbp1 expression plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 7674) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7674 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 96..475 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 476..679 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 630..650 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 724..743 /label=T7 /note="T7 promoter, forward primer" promoter 724..742 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2353..3072 /codon_start=1 /product="cyan variant of GFP" /label=CFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2398..2419) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2659..2678) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3006..3027 /label=EGFP-C /note="EGFP, forward primer" promoter complement(3088..3106) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(3089..3106) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(3126..3143) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3132..3356 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3402..3830 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3489..3508) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3699..3720 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3839..3859) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3844..4173 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4024..4159 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4086..4105 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4240..5034 /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(4294..4313) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4904..4923 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5208..5329 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5245..5264) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5299..5318 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5378..5394) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5378..5394) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5391..5413) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5402..5418 /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(5426..5456) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5471..5492 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5609..5626) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5780..6368) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5860..5879) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6539..7399) /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 7162..7181 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7400..7504) /gene="bla" /label=AmpR promoter primer_bind complement(7579..7598) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc 61 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta 121 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 181 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 241 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 301 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 361 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 421 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat 481 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag 541 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc 601 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga 661 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga 721 aattaatacg actcactata gggagaccca agcttggtac cgagctcgga tccactagta 781 acggccgcca gtgtgctgga attcaccatg atcagctcag actcgaggtc ctcccctggc 841 ttggctcggt gggctgagag ctatgaggcc aagtgtgagc gacgtcagga gacccgagaa 901 aaccgtcgcc gccgccgcaa tgagaccact tgccgccagc cagggaaggt gctgaggacc 961 caacacaagg aacggcttca gggagctcgg cagctacagt tcctcaaaag gagaaacctg 1021 gaagaagaga agaaaggcca agccagggaa caagggccct ccagcaggac agatggagga 1081 acaggccaag tgtctatcct caaggagtcc ttgcctggtg ccaacaaggc ctctttccct 1141 ggacagcagg aaacagggat aagctctgag gttttcccgg ccttacatca ttcctcctca 1201 ggcatccaga gggatctggg tggccaccat gcctcccatg ggagagcctt tccaccacag 1261 gactctgaca tcaagaagcc acacagacag caccggggca ctcagacaaa ggcagaagag 1321 gcattgccaa caataaagaa tgatgccagt cagcaaacca aatgtggagt tgctgtccta 1381 gacaaggaca tcattcaact ctcggaatac ctcaaagaag ccttacatag ggagctgatc 1441 ctaaaaaaga aaatggtgat tctccaggac ctcctgcctg ccctgatccg ggcctctgac 1501 agctcctgga agggacagct taatgaagac aagctgaagg gcaaactaag atccctagaa 1561 aaccagctct acacctgcct gcagaaacac tccccctggg gaatgaaaaa ggtgctgctg 1621 gagatagaag accagagaag cagctatgag cagaaggcta aggcgtcact gcagaaagtg 1681 ctggaggaga aaatgtgtgc agagcagcag ctacagaggg cacagctgtc cctggctttg 1741 gcagagcaga agtgtcagga gtggaagagt cagtacgagg ctctgaagga ggactggagg 1801 accctagggg accagcacag agagctggag agccaactgc atgtgcttca gtccaaacta 1861 cagggagcag acagcagaga cccacagatg agccaggccc tgcagctttt agaaaatgag 1921 caccaggagc tgcagaccaa actcgagagc ctgcagggag acggagagca gcagagctct 1981 gagacccagg acctacaaga tcaactaaag aagtcagagg aagagaaaca ggccctggtg 2041 agcaaagtac aacagctaca gagtctgctt cagaatcaat ccttacaact tcaagaacag 2101 gagaagctct taaagaaaga tcagggtctg cctgtgtgga atccaaagct ctcccttgat 2161 gaagtgaagc ctgagggcac aagaaaggag aaagaggagg agctcagaga ccaactacaa 2221 aaggagacat ttcagctcca agtcaaggaa aaggagctgc agtgtggtca atggctccca 2281 gtgctgatgg tggtgattgc tacagcactg gcagtgttcc tgggaaacaa aggcaacctg 2341 gtcatcgtcg agatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 2401 gtcgagctgg acggcgacgt aaacggccac aggttcagcg tgtccggcga gggcgagggc 2461 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 2521 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc 2581 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 2641 cgtaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 2701 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 2761 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac 2821 aagcagaaga acggcatcaa ggcccacttc aagatccgcc acaacatcga ggacggcagc 2881 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 2941 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 3001 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 3061 ctgtacaagt aatctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg 3121 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 3181 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 3241 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 3301 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 3361 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 3421 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 3481 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 3541 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 3601 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 3661 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 3721 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 3781 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 3841 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 3901 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 3961 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 4021 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 4081 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 4141 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 4201 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg 4261 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag 4321 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt 4381 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta 4441 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg 4501 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt 4561 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat 4621 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg 4681 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca 4741 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc 4801 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc 4861 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat 4921 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc 4981 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga 5041 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt 5101 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga 5161 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg 5221 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt 5281 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta 5341 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 5401 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 5461 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 5521 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 5581 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 5641 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 5701 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 5761 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 5821 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 5881 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 5941 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 6001 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 6061 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 6121 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 6181 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 6241 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 6301 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 6361 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 6421 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 6481 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 6541 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 6601 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 6661 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 6721 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 6781 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 6841 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 6901 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 6961 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 7021 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 7081 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 7141 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 7201 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 7261 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 7321 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 7381 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 7441 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 7501 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc 7561 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct 7621 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctac //