LOCUS Exported 6516 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMSCV-pBabeMCS-IRES-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6516) TITLE Sherr/Roussel lab unpublished plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 6516) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6516 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(592..611) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind complement(739..761) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(955..974) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1074..1096 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1134..1152) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1220..1324 /gene="bla" /label=AmpR promoter CDS 1325..2185 /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(1543..1562) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2356..2944 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2845..2864 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3098..3115 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3232..3253 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3268..3298 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3306..3322 /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)." LTR 3484..3998 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" CDS complement(4160..4879) /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS complement(4163..4879) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4205..4226) /label=EGFP-C /note="EGFP, forward primer" primer_bind 4554..4573 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4813..4834 /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5041..5060) /label=IRES-F /note="IRES internal ribosome entry site, forward primer" primer_bind 5270..5287 /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" CDS complement(5504..5920) /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind complement(5520..5536) /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(5552..5574) /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" misc_feature 5987..6328 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" LTR 6392..392 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 gtctatcgga ggactggcgc gccgagtgag gggttgtggg ctcttttatt gagctcgggg 61 agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa ataaggcaca 121 gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta gaaactgctg 181 agggcgggac cgcatctggg gaccatctgt tcttggccct gagccggggc aggaactgct 241 taccacagat atcctgtttg gcccatattc tgctgtctct ctgttcctaa ccttgatctg 301 aacttctcta ttctcagtta tgtattttcc atgccttgca aaatggcgtt acttaagcta 361 gcttgccaaa cctacaggtg gggtctttca ttcccccctt tttctggaga ctaaataaaa 421 tcttttattt tatctatggc tcgtactcta taggcttcag ctggtgatat tgttgagtca 481 aaactagagc ctggaccact gatatcctgt ctttaacaaa ttggactaat cgcctctacg 541 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg 601 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg 661 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg 721 caccattcct tgcgccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca 781 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg 841 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta 901 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat 961 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc 1021 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag 1081 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt 1141 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg 1201 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 1261 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 1321 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 1381 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 1441 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 1501 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 1561 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 1621 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 1681 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 1741 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 1801 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 1861 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 1921 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 1981 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 2041 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 2101 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 2161 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 2221 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 2281 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 2341 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 2401 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 2461 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 2521 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 2581 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 2641 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 2701 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 2761 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 2821 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 2881 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 2941 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 3001 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 3061 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc 3121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag 3181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag 3241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg 3301 tggaattgtg agcggataac aatttcacaa ttctccccag ctctttctgc gctgtccata 3361 atgaactatt tcaggagtat tggagtatgt ggattcaaaa cagtggctaa ttgagtcagg 3421 attacttaat attcgactcc taccctcaga atgttgttct ccaacttcaa gaaactgtta 3481 cccatgaaag acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac 3541 agcgagacca caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg 3601 cgacgcagtc tatcggagga ctggcgcgcc gagtgagggg ttgtgggctc ttttattgag 3661 ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata 3721 aggcacaggg tcatttcagg tccttggggc accctggaaa catctgatgg ttctctagaa 3781 actgctgagg gcgggaccgc atctggggac catctgttct tggccctgag ccggggcagg 3841 aactgcttac cacagatatc ctgtttggcc catattctgc tgtctctctg ttcctaacct 3901 tgatctgaac ttctctattc tcagttatgt attttccatg ccttgcaaaa tggcgttact 3961 taagctagct tgccaaacct acaggtgggg tctttcattc cccccttttt ctggagacta 4021 aataaaatct tttattttat cgatatcagc gctttaaatt tgcgcatgct agctatagtt 4081 ctagaggtac cggttgttaa cgttagccgg ctacctgaag cttgcatgcc tgcaggtcga 4141 ctctagagtc gcggccgctt tacttgtaca gctcgtccat gccgagagtg atcccggcgg 4201 cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcaggg 4261 cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg 4321 tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg tggcggatct 4381 tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg ttgtggctgt 4441 tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct 4501 tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt 4561 agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg ggcatggcgg 4621 acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt 4681 aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga 4741 acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac acgctgaact 4801 tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct 4861 cctcgccctt gctcaccatg gtattatcgt gtttttcaaa ggaaaaccac gtccccgtgg 4921 ttcggggggc ctagacgttt ttttaacctc gactaaacac atgtaaagca tgtgcaccga 4981 ggccccagat cagatcccat acaatggggt accttctggg catccttcag ccccttgttg 5041 aatacgcttg aggagagcca tttgactctt tccacaacta tccaactcac aacgtggcac 5101 tggggttgtg ccgcctttgc aggtgtatct tatacacgtg gcttttggcc gcagaggcac 5161 ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt 5221 caagaagctt ccagaggaac tgcttccttc acgacattca acagaccttg cattcctttg 5281 gcgagagggg aaagacccct aggaatgctc gtcaagaaga cagggccagg tttccgggcc 5341 ctcacattgc caaaagacgg caatatggtg gaaaataaca tatagacaaa cgcacaccgg 5401 ccttattcca agcggcttcg gccagtaacg ttaggggggg ggcggaattg atcccgctcg 5461 agatctgaat tcctacgtac caccacactg ggatcccaat tccggcgcct agagaaggag 5521 tgagggctgg ataaagggag gatcgaggcg gggtcgaacg aggaggttca agggggagag 5581 acggggcgga tggaggaaga ggaggcggag gcttagggtg tacaaagggc ttgacccagg 5641 gaggggggtc aaaagccaag gcttcccagg tcacgatgta ggggacctgg tctgggtgtc 5701 catgcgggcc aggtgaaaag accttgatct taacctgggt gatgaggtct cggttaaagg 5761 tgccgtctcg cggccatccg acgttaaagg ttggccattc tgcagagcag aaggtaaccc 5821 aacgtctctt cttgacatct accgactggt tgtgagcgat ccgctcgaca tctttccagt 5881 gacctaaggt caaacttaag ggagtggtaa cagtctggcc ctaattttca gacaaataca 5941 gaaacacagt cagacagaga caacacagaa cgatgctgca gcgctgcagc agacaagacg 6001 cgcggcttcg gttccaaacc gaaagcaaaa attcagacgg aggcgggaac tgttttaggt 6061 tctcgtctcc taccagaacc acatatcctg acggggtcgg attccacatc gactcccttc 6121 ctcaggtcgg gccacaaaaa cggcccccaa agtccctggg acgtctccca gggttgcggc 6181 cgggtgttca gaactcgtca gttccaccac gggtccgcca gatacagagc tagttagcta 6241 actagtacag acgcaggcgc aaacattaga tgccggcaca aacacgcaca aagacagaga 6301 cagacacgaa acgaccgctg gccagcttac ctcccggcgg gggggtcggt ggtccctggg 6361 caggggtctc caaatctcgg tggaacctcc aaatgaaaga cccccgaggt gggcagtcaa 6421 tcaatctgag gagaccctcc caaggatcag cgagtccacg attcggatgc aaacagcaag 6481 aggctttatt gggaatacgg gtacccgggc gacgca //