LOCUS Exported 8957 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pLenti CMV GSE T329A Hygro (w189-4) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8957) TITLE shMDC1 plasmids REFERENCE 2 (bases 1 to 8957) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8957 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 27..131 /gene="bla" /label=AmpR promoter CDS 132..992 /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(350..369) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1163..1751 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1652..1671 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1905..1922 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2039..2060 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2075..2105 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2113..2129 /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 2118..2140 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2137..2153 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2137..2153 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2172..2192 /label=T3 /note="T3 promoter, forward primer" promoter 2174..2192 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" LTR 2448..2628 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 2672..2797 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3290..3523 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 3707..3751 /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 4012..4129 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 4152..4455 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4456..4659 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4610..4630 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4656..4680 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 4764..4788 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(5082..5106) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_feature 5137..5725 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5190..5210) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5608..5619) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" promoter 5781..6280 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(5816..5839) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 6213..6232 /label=mPGK-F /note="Mouse PGK promoter, forward primer" LTR 7698..7931 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 8003..8124 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8040..8059) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8094..8113 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 8143..8278 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8205..8224 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(8310..8329) /label=T7 /note="T7 promoter, forward primer" promoter complement(8311..8329) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8339..8356) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(8339..8355) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8348..8370) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 8500..8955 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8587..8606) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8797..8818 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta 2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat 2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc 2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg 2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc 2401 gcattgcaga gatattgtat ttaagtgcct agctcgatac aataaacggg tctctctggt 2461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 2521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 2581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa 2641 cagggacctg aaagcgaaag ggaaaccaga gctctctcga cgcaggactc ggcttgctga 2701 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag 2761 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag 2821 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca 2881 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac 2941 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga 3001 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga 3061 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac 3121 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt 3181 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca 3241 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt 3301 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg 3361 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta 3421 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa 3481 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct 3541 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc 3601 tggaacagat tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac 3661 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga 3721 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct 3781 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt 3841 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac 3901 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga 3961 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggttaa cttttaaaag 4021 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga 4081 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta tcgataagct 4141 tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 4201 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 4261 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 4321 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 4381 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 4441 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt 4501 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac 4561 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc 4621 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc 4681 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccgact ctagtccagt 4741 gtggtggaat tctgcagata tcaacaagtt tgtacaaaaa agcaggcttt aaaggaacca 4801 attcagtcga ctggatccac catgtcctct ccccagcaaa agaaaaaacc actcgatgga 4861 gaatatttcg cccttaagat ccgtgggcgt gagcgcttcg agatgttccg agagctgaat 4921 gaggccttgg aattaaagga tgcccgtgct gccgaggagt caggagacag cagggctcac 4981 tccagctacc cgaagaccaa gaagggccag tctacgtccc gccataaaaa accaatgatc 5041 aagaaagtgg ggcctgactc agactgagtc gagatatcta gacccagctt tcttgtacaa 5101 agtggttgat atccagcaca gtggcggccg ctcgacaatc aacctctgga ttacaaaatt 5161 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct 5221 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg 5281 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc 5341 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt 5401 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc 5461 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg 5521 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg 5581 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc 5641 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg 5701 atctcccttt gggccgcctc cccgcctgga attctgcaga tatccggact agtgatctct 5761 cgaggttaac gaattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg 5821 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca 5881 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc 5941 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac 6001 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca 6061 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg 6121 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg 6181 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc 6241 tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcatcc cgccaccatg 6301 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc 6361 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta 6421 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt 6481 tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg 6541 gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa 6601 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg 6661 atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc 6721 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac 6781 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg 6841 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc 6901 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg 6961 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt 7021 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg 7081 ctccggggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg 7141 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc 7201 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg 7261 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga 7321 atagagtaga tgccgaccga acaagagctg atttcgagaa cgcctcagcc agcaactcgc 7381 gcgagcctag caaggcaaat gcgagagaac ggccttacgc ttggtggcac agttctcgtc 7441 cacagttcgc taagctcgct cggctgggtc gcgggagggc cggtcgcagt gattcaggcc 7501 cttctggatt gtgttggtcc ccagggcacg attgtcatgc ccacgcactc gggtgatctg 7561 actgatcccg cagattggag atcgccgccc gtgcctgccg attgggtgca gatccgtcga 7621 gggcccggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt 7681 aaaagaaaag gggggactgg aagggctagc tcactcccaa cgaagacaag atctgctttt 7741 tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctgcct 7801 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc 7861 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa 7921 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa 7981 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag 8041 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 8101 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg 8161 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 8221 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 8281 ggcttttgcg tcgagacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac 8341 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc 8401 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc 8461 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat 8521 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 8581 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 8641 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 8701 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 8761 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 8821 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 8881 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 8941 taacgtttac aatttcc //