LOCUS Exported 6468 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION a bicistronic IRES-DsRed containing retroviral vector, MCS is expanded from pMSCV-IRES-GFP. ACCESSION . VERSION . KEYWORDS pMSCV-IRES-DsRed FP (pMIDsRed II) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6468) AUTHORS Holst J, Szymczak-Workman AL, Vignali KM, Burton AR, Workman CJ, Vignali DA TITLE Generation of T-cell receptor retrogenic mice. JOURNAL Nat Protoc. 2006;1(1):406-17. PUBMED 17406263 REFERENCE 2 (bases 1 to 6468) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6468 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 169..510 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 577..993 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 923..945 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 961..977 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(1206..1223) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1433..1452 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 1614..2291 /codon_start=1 /product="rapidly maturing tetrameric variant of DsRed fluorescent protein (Bevis and Glick, 2002)" /label=DsRed-Express /note="mammalian codon-optimized" /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH LFL" primer_bind complement(1744..1762) /label=mCherry-R /note="mCherry, reverse primer" primer_bind complement(1794..1814) /label=DsRed1-N /note="DsRed1, reverse primer" primer_bind 2012..2031 /label=mCherry-F /note="mCherry, forward primer" primer_bind 2206..2229 /label=DsRed1-C /note="DsRed1, forward primer" LTR 2451..2965 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 3127..3143 /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(3151..3181) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3196..3217 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3334..3351) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3505..4093) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3585..3604) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4264..5124) /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 4887..4906 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5125..5229) /gene="bla" /label=AmpR promoter primer_bind 5297..5315 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5353..5375) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5475..5494 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5688..5710 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 5838..5857 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 6057..105 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" ORIGIN 1 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag 61 ggtctcctca gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga 121 gatttggaga cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc 181 ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc 241 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac 301 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt 361 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt 421 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt 481 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc 541 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta 601 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat 661 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga 721 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt 781 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg 841 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt 901 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc 961 ctttatccag ccctcactcc ttctctaggc gccggaattc agatcttacg tagctagcgg 1021 atcccaattg ctcgagcggg atcaattccg ccccccccct aacgttactg gccgaagccg 1081 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt 1141 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct 1201 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct 1261 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc 1321 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc 1381 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc 1441 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc 1501 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct 1561 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat accatggcct 1621 cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga 1681 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga 1741 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc 1801 cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca 1861 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg 1921 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg ctccttcatc tacaaggtga 1981 agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct 2041 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca 2101 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagtcc atctacatgg 2161 ccaagaagcc cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct 2221 cccacaacga ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc 2281 tgttcctgta gcggccgcga ctctagagtc gacctgcagg catgcaagct tcaggtagcc 2341 ggctaacgtt aacaaccggt acctctagaa ctatagctag catgcgcaaa tttaaagcgc 2401 tgatatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc 2461 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac 2521 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg 2581 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt 2641 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc 2701 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc 2761 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg 2821 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt 2881 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga 2941 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc 3001 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac 3061 atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt 3121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 3181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 3241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 3301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 3361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 3421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 3481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 3541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 3601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 3661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 3721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 3781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 3841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 3901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 3961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 4021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 4081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga 4141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct 4201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga 4261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc 4321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg 4381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat 4441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat 4501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg 4561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc 4621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa 4681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc 4741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt 4801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag 4861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt 4921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag 4981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac 5041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc 5101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca 5161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg 5221 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat 5281 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga 5341 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc 5401 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg 5461 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga 5521 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct 5581 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 5641 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 5701 ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg 5761 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga 5821 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg 5881 ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac 5941 aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata 6001 gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg 6061 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg 6121 aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga 6181 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac 6241 agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc 6301 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc 6361 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct 6421 cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgt //