LOCUS Exported 8809 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expression in mammalian cells. ACCESSION . VERSION . KEYWORDS pCL20c-MSCV-IRES-CRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8809) TITLE Roussel lab plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 8809) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8809 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..342 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 409..825 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 755..777 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 793..809 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(1042..1059) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1269..1288 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 1474..2502 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED GD" primer_bind complement(2753..2772) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2818..2873 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2872..2891) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3234..3250) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3234..3250) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3247..3269) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3258..3274 /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(3282..3312) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3327..3348 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3407..3602 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3453..3588 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3515..3534 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 3608..3742 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3645..3664) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3699..3718 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3810..3827) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3981..4569) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4061..4080) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4740..5600) /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 5363..5382 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5601..5705) /gene="bla" /label=AmpR promoter enhancer 5736..6115 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" LTR 6117..6374 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 6421..6546 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 7038..7155 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" misc_feature 7515..7748 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 7933..7977 /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" LTR 8230..8746 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc 61 taatgtttgc gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt 121 ggtggaactg acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt 181 gggggccgtt tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca 241 ggatatgtgg ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt 301 ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt 361 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt 421 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac 481 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc 541 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag 601 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc 661 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct 721 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg 781 cctcgatcct ccctttatcc agccctcact ccttctctag gcgccggaat tgggatccca 841 gtgtggtggt acgtaggaat tcagatctcg agcgggatca attccgcccc ccccctaacg 901 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca 961 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga 1021 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga 1081 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca 1141 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag 1201 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa 1261 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac 1321 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga 1381 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac 1441 gataatacca tggccaagaa gaagaggaag gtgtccaatt tactgaccgt acaccaaaat 1501 ttgcctgcat taccggtcga tgcaacgagt gatgaggttc gcaagaacct gatggacatg 1561 ttcagggatc gccaggcgtt ttctgagcat acctggaaaa tgcttctgtc cgtttgccgg 1621 tcgtgggcgg catggtgcaa gttgaataac cggaaatggt ttcccgcaga acctgaagat 1681 gttcgcgatt atcttctata tcttcaggcg cgcggtctgg cagtaaaaac tatccagcaa 1741 catttgggcc agctaaacat gcttcatcgt cggtccgggc tgccacgacc aagtgacagc 1801 aatgctgttt cactggttat gcggcggatc cgaaaagaaa acgttgatgc cggtgaacgt 1861 gcaaaacagg ctctagcgtt cgaacgcact gatttcgacc aggttcgttc actcatggaa 1921 aatagcgatc gctgccagga tatacgtaat ctggcatttc tggggattgc ttataacacc 1981 ctgttacgta tagcagaaat tgccaggatc agggttaaag atatctcacg tactgacggt 2041 gggagaatgt taatccatat tggcagaacg aaaacgctgg ttagcaccgc aggtgtagag 2101 aaggcactta gcctgggggt aactaaactg gtcgagcgat ggatttccgt ctctggtgta 2161 gctgatgatc cgaataacta cctgttttgc cgggtcagaa aaaatggtgt tgccgcgcca 2221 tctgccacca gccagctatc aactcgcgcc ctggaaggga tttttgaagc aactcatcga 2281 ttgatttacg gcgctaagga tgactctggt cagagatacc tggcctggtc tggacacagt 2341 gcccgtgtcg gagccgcgcg agatatggcc cgcgctggag tttcaatacc ggagatcatg 2401 caagctggtg gctggaccaa tgtaaatatt gtcatgaact atatccgtaa cctggatagt 2461 gaaacagggg caatggtgcg cctgctggaa gatggcgatt aggcggccgc atcgatgccg 2521 tatacggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta 2581 aaagaaaagg ggggactgga agggctaatt cactcccaaa gaagacaaga tctgcttttt 2641 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta 2701 gggaacccac tgcttaagcc tcaataaagc ttcagctgct cgagctagca gatctttttc 2761 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat 2821 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa 2881 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg 2941 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt 3001 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt 3061 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct 3121 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc 3181 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag 3241 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc 3301 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc 3361 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt 3421 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 3481 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 3541 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca 3601 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa 3661 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa 3721 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc 3781 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 3841 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 3901 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 3961 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 4021 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 4081 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 4141 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 4201 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 4261 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 4321 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 4381 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 4441 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 4501 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 4561 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 4621 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 4681 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 4741 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 4801 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 4861 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 4921 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 4981 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 5041 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 5101 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 5161 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 5221 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 5281 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 5341 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 5401 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 5461 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 5521 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 5581 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 5641 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 5701 ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattga ctagttatta 5761 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 5821 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 5881 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 5941 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 6001 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 6061 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgggagg 6121 cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct 6181 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 6241 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 6301 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 6361 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag 6421 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga 6481 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag 6541 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca 6601 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga 6661 ttcgcagtta atactggcct gttagaaaca tcagaaggct gtagacaaat actgggacag 6721 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca 6781 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag 6841 atagaggaag agcaaaacaa aagtaagaaa aaagcacagc aagcagcagg atcttcagac 6901 ctggaaattc cctacaatcc ccaaagtcaa ggagtagtag aatctatgaa taaagaatta 6961 aagaaaatta taggacaggt aagagatcag gctgaacatc ttaagacagc agtacaaatg 7021 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 7081 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 7141 aaaattcaaa attttcgggt ttattacagg gacagcagaa atccactttg gaaaggacca 7201 gcaaagctcc tctggaaagg tgaaggggca gtagtaatac aagataatag tgacataaaa 7261 gtagtgccaa gaagaaaagc aaagatcatt agggattatg gaaaacagat ggcaggtgat 7321 gattgtgtgg caagtagaca ggatgaggat tagaacatgg aaaagtttag taaaacacca 7381 taaggaggag atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa 7441 attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa 7501 agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg 7561 ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag 7621 cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc 7681 tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa 7741 cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg 7801 aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag 7861 tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa 7921 aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg 7981 aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga 8041 ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag 8101 ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc gagctcaagc 8161 ttcgaacgcg ttaacgggcc cagcttcgat aaaataaaag attttattta gtctccagaa 8221 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt 8281 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca 8341 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct 8401 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc 8461 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa 8521 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga 8581 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg 8641 tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga 8701 gggtctcctc agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg 8761 agatttggag acccctgccc agggaccacc gacccccccg ccgggaggt //