LOCUS Exported 6512 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION a bicistronic IRES-GFP containing retroviral vector, MCS is expanded from pMSCV-IRES-GFP. ACCESSION . VERSION . KEYWORDS pMSCV-IRES-GFP II (pMIG 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" LTR 1..517 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 581..922 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 989..1405 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1335..1357 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1373..1389 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(1618..1635) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1845..1864 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2026..2745 /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 2026..2742 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2071..2092) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2332..2351) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2679..2700 /label=EGFP-C /note="EGFP, forward primer" LTR 2907..3421 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 3583..3599 /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(3607..3637) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3652..3673 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3790..3807) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3961..4549) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4041..4060) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4720..5580) /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 5343..5362 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5581..5685) /gene="bla" /label=AmpR promoter primer_bind 5753..5771 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5809..5831) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5931..5950 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6144..6166 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6294..6313 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc 61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag 121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa 181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt 241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt 301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac 361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa 421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct 481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga 541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt 601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct 661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg 721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc 781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg 841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg 901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac 961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac 1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa 1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg 1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg 1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga 1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc 1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc 1381 agccctcact ccttctctag gcgccggaat tcagatctta cgtagctagc ggatcccaat 1441 tgctcgagcg ggatcaattc cgcccccccc ctaacgttac tggccgaagc cgcttggaat 1501 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg 1561 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc 1621 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt 1681 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg 1741 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac 1801 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg 1861 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg 1921 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc 1981 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatggt gagcaagggc 2041 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 2101 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 2161 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 2221 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc 2281 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 2341 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 2401 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 2461 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac 2521 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag 2581 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag 2641 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 2701 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct 2761 agagtcgacc tgcaggcatg caagcttcag gtagccggct aacgttaaca accggtacct 2821 ctagaactat agctagcatg cgcaaattta aagcgctgat atcgataaaa taaaagattt 2881 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag 2941 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag 3001 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc 3061 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag 3121 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt 3181 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc 3241 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc 3301 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct 3361 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca 3421 tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag 3481 taatcctgac tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt 3541 tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat 3601 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 3661 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg 3721 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc 3781 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc 3841 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa 3901 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt 3961 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg 4021 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg 4081 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag 4141 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc 4201 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa 4261 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg 4321 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc 4381 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac 4441 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 4501 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 4561 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt 4621 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa 4681 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga 4741 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt 4801 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg 4861 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga 4921 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga 4981 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg 5041 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc 5101 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc 5161 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca 5221 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac 5281 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg 5341 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc 5401 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg 5461 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac 5521 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat 5581 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 5641 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 5701 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 5761 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 5821 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 5881 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 5941 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 6001 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 6061 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 6121 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat 6181 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc 6241 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg 6301 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt 6361 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta 6421 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa 6481 aagattttat ttagtctcca gaaaaagggg gg //