LOCUS Exported 6512 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION a bicistronic IRES-CFP containing retroviral vector, MCS is expanded from pMSCV-IRES-GFP. ACCESSION . VERSION . KEYWORDS pMSCV-IRES-CFP II (pMIC II) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6512) 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 6512) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6512 /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" primer_bind complement(1616..1633) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1843..1862 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2024..2743 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2069..2090) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2330..2349) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2677..2698 /label=EGFP-C /note="EGFP, forward primer" LTR 2905..3419 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 3581..3597 /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(3605..3635) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3650..3671 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3788..3805) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3959..4547) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4039..4058) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4718..5578) /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 5341..5360 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5579..5683) /gene="bla" /label=AmpR promoter primer_bind 5751..5769 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5807..5829) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5929..5948 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6142..6164 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6292..6311 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 6511..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 gccggaattc agatcttacg tagctagcgg atcccaattg 1441 ctcgagcggg atcaattccg ccccccccct aacgttactg gccgaagccg cttggaataa 1501 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg 1561 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc 1621 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct 1681 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac 1741 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc 1801 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta 1861 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg 1921 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg 1981 aaccacgggg acgtggtttt cctttgaaaa acacgataat accatggtga gcaagggcga 2041 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca 2101 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa 2161 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac 2221 ctggggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa 2281 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa 2341 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct 2401 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta 2461 catcagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt 2521 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa 2581 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc 2641 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac 2701 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag 2761 agtcgacctg caggcatgca agcttcaggt agccggctaa cgttaacaac cggtacctct 2821 agaactatag ctagcatgcg caaatttaaa gcgctgatat cgataaaata aaagatttta 2881 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct 2941 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat 3001 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag 3061 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca 3121 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc 3181 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg 3241 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt 3301 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg 3361 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg 3421 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta 3481 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc 3541 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc 3601 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 3661 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 3721 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 3781 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 3841 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 3901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 3961 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 4021 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 4081 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 4141 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 4201 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 4261 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 4321 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 4381 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 4441 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 4501 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 4561 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 4621 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 4681 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 4741 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 4801 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 4861 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 4921 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 4981 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 5041 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 5101 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 5161 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 5221 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 5281 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 5341 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 5401 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 5461 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 5521 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 5581 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 5641 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 5701 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta 5761 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc 5821 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc 5881 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc 5941 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa 6001 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 6061 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa 6121 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg caaggaatgg 6181 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac 6241 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc 6301 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc 6361 ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc aggctctagt 6421 tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga taaaataaaa 6481 gattttattt agtctccaga aaaagggggg aa //