LOCUS Exported 7871 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Retroviral plasmid containing tdTomato for labeling of cells. ACCESSION . VERSION . KEYWORDS pMSCVpuro-tdTomato SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7871) AUTHORS Kleeman B, Olsson A, Newkold T, Kofron M, DeLay M, Hildeman D, Grimes HL TITLE A guide to choosing fluorescent protein combinations for flow cytometric analysis based on spectral overlap. JOURNAL Cytometry A. 2018 Mar 13. doi: 10.1002/cyto.a.23360. PUBMED 29533508 REFERENCE 2 (bases 1 to 7871) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7871 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 579..920 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 987..1403 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1333..1355 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1371..1387 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" misc_feature 1410..1466 /label=MCS /note="multiple cloning site" regulatory 1474..1483 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1480..2910 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" promoter 3045..3544 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(3080..3103) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 3477..3496 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 3565..4164 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3565..3584) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4061..4081 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 4264..4778 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 4940..4956 /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(4964..4994) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5009..5030 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5147..5164) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5318..5906) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5398..5417) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6077..6937) /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 6700..6719 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6938..7042) /gene="bla" /label=AmpR promoter primer_bind 7110..7128 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7166..7188) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7288..7307 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7501..7523 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7651..7670 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 7870..515 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca 121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga 181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt 241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc 301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc 361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata 421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca 481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga 541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg 601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt 661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa 721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga 781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag 841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa 901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg 961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct 1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga 1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag 1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc 1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc 1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg 1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag 1381 ccctcactcc ttctctaggc gccggaatta gatctcgagc tcaagcttcg aattctgcag 1441 tcgacggtac cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag 1501 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag 1561 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg 1621 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg 1681 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc 1741 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc 1801 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc 1861 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc 1921 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag 1981 ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc 2041 gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag 2101 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgggg 2161 catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga ggacaacaac 2221 atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc 2281 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc 2341 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag 2401 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag 2461 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg 2521 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg 2581 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag 2641 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc 2701 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag 2761 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac 2821 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc 2881 ctgtacggca tggacgagct gtacaagtaa gcggccgcga ctctagatca taatcagcca 2941 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct 3001 gaaacataaa atgaatgcaa ttgttgttgt taacgaattc taccgggtag gggaggcgct 3061 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac 3121 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt 3181 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc 3241 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt 3301 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata 3361 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg 3421 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt 3481 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct 3541 ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc 3601 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 3661 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 3721 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 3781 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 3841 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 3901 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 3961 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 4021 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 4081 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 4141 atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg 4201 agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca gaaaaagggg 4261 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag 4321 gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga acagagagac 4381 agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc 4441 aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat 4501 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca 4561 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca 4621 acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtgtatcc 4681 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc 4741 tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt cttgaagttg 4801 gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca attagccact 4861 gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag cgcagaaaga 4921 gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa 4981 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc 5041 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc 5101 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact 5161 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 5221 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 5281 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 5341 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 5401 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 5461 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 5521 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 5581 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 5641 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 5701 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa 5761 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 5821 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 5881 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 5941 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 6001 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 6061 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 6121 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 6181 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 6241 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 6301 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 6361 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 6421 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 6481 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 6541 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 6601 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 6661 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 6721 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 6781 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 6841 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 6901 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 6961 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 7021 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa 7081 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg 7141 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag 7201 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg 7261 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc 7321 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt 7381 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac 7441 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt 7501 cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg agatggcgcc 7561 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag 7621 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac 7681 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc gattagtcca 7741 atttgttaaa gacaggatat cagtggtcca ggctctagtt ttgactcaac aatatcacca 7801 gctgaagcct atagagtacg agccatagat aaaataaaag attttattta gtctccagaa 7861 aaagggggga a //