LOCUS Exported 8958 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pLenti CMV RasV12 Neo (w108-1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8958) TITLE shMDC1 plasmids REFERENCE 2 (bases 1 to 8958) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8958 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 48..67 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 129..932 /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(192..211) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 802..821 /label=Neo-F /note="Neomycin resistance gene, forward primer" LTR 1023..1256 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 1328..1449 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1365..1384) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1419..1438 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 1489..1624 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1551..1570 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(1644..1663) /label=T7 /note="T7 promoter, forward primer" promoter complement(1645..1663) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1673..1690) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1673..1689) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1682..1704) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 1831..2286 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1918..1937) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2128..2149 /label=F1ori-F /note="F1 origin, forward primer" promoter 2312..2416 /gene="bla" /label=AmpR promoter CDS 2417..3277 /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(2635..2654) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3448..4036 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3937..3956 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4190..4207 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4324..4345 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4360..4390 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4398..4414 /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)." primer_bind 4403..4425 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4422..4438 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4422..4438 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 4457..4477 /label=T3 /note="T3 promoter, forward primer" promoter 4459..4477 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 4505..4731 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 4732..4912 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 4959..5084 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 5577..5810 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 5995..6039 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 6306..6422 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 6445..6748 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6749..6952 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6903..6923 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 6949..6973 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 7057..7081 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 7156..7725 /codon_start=1 /product="human oncoprotein generated by the G12V mutation in the small GTPase H-Ras" /label=H-Ras (G12V) /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI RQHKLRKLNPPDESGPGCMSCKCVLS" protein_bind complement(7888..7912) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 7943..8531 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7996..8016) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8414..8425) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" promoter 8574..115 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(8609..8632) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" ORIGIN 1 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct 61 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg 121 cagccaatat gggatcggcc attgaacaag atggattgca cgcaggttct ccggccgctt 181 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg 241 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg 301 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg 361 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg 421 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca 481 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc 541 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc 601 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca 661 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga 721 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg 781 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg 841 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg 901 ccttctatcg ccttcttgac gagttcttct gaggggatcc gtcgagggcc cggtaccttt 961 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg 1021 actggaaggg ctagctcact cccaacgaag acaagatctg ctttttgctt gtactgggtc 1081 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct 1141 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga 1201 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag 1261 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag 1321 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 1381 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 1441 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta 1501 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 1561 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 1621 ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg 1681 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag 1741 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc 1801 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg 1861 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 1921 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 1981 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 2041 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 2101 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 2161 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 2221 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta 2281 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 2341 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 2401 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc 2461 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga 2521 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct 2581 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg 2641 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta 2701 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat 2761 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt 2821 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga 2881 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga 2941 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga 3001 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc 3061 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc 3121 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg 3181 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat 3241 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata 3301 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 3361 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 3421 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 3481 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 3541 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 3601 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 3661 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 3721 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 3781 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 3841 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 3901 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 3961 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 4021 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 4081 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 4141 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt 4201 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat 4261 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc 4321 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc 4381 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca 4441 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc 4501 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa 4561 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac 4621 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa 4681 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc 4741 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 4801 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 4861 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc 4921 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc 4981 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt 5041 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga 5101 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat 5161 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt 5221 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag 5281 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa 5341 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa 5401 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg 5461 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta 5521 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga 5581 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg 5641 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg 5701 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc 5761 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg 5821 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat 5881 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac 5941 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat 6001 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca 6061 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga 6121 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg 6181 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa 6241 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcgg 6301 ttaactttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac 6361 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa attcaaaatt 6421 ttatcgataa gcttgggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg 6481 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc 6541 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc 6601 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg 6661 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat 6721 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg 6781 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag 6841 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt 6901 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt 6961 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg 7021 actctagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa aaaagcaggc 7081 tttaaaggaa ccaattcagt cgactggatc ccagtgtggt ggtacagctt ggggggccag 7141 ggccggggcc gagcgatgac ggaatataag ctggtggtgg tgggcgccgt cggtgtgggc 7201 aagagtgcgc tgaccatcca gctgatccag aaccattttg tggacgaata cgaccccact 7261 atagaggatt cctaccggaa gcaggtggtc attgatgggg agacgtgcct gttggacatc 7321 ctggataccg ccggccagga ggagtacagc gccatgcggg accagtacat gcgcaccggg 7381 gagggcttcc tgtgtgtgtt tgccatcaac aacaccaagt cttttgagga catccaccag 7441 tacagggagc agatcaaacg ggtgaaggac tcggatgacg tgcccatggt gctggtgggg 7501 aacaagtgtg acctggctgc acgcactgtg gaatctcggc aggctcagga cctcgcccga 7561 agctacggca tcccctacat cgagacctcg gccaagaccc ggcagggagt ggaggatgcc 7621 ttctacacgt tggtgcgtga gatccggcag cacaagctgc ggaagctgaa ccctcctgat 7681 gagagtggcc ccggctgcat gagctgcaag tgtgtgctct cctgacgcag cacaagctca 7741 ggacatggag gtgccggatg caggaaggag gtgcagacgg aaggaggagg aaggaaggac 7801 ggaagcaagg aaggaaggaa gggctgctgg agcccagtca ccccccaagc tgtaggaatt 7861 cgccagcaca gtggtcgaga tatctagacc cagctttctt gtacaaagtg gttgatatcc 7921 agcacagtgg cggccgctcg acaatcaacc tctggattac aaaatttgtg aaagattgac 7981 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt 8041 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt 8101 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt 8161 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg 8221 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg 8281 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct 8341 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt 8401 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc 8461 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc 8521 cgcctccccg cctggaattc tgcagatatc cggactagtg atctaattct accgggtagg 8581 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg 8641 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc 8701 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt 8761 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac 8821 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag 8881 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc 8941 cgggggcggg ctcagggg //