LOCUS Exported 6512 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION a bicistronic IRES-YFP containing retroviral vector, MCS is expanded from pMSCV-IRES-GFP. ACCESSION . VERSION . KEYWORDS pMSCV-IRES-YFP II (pMIY 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" primer_bind 102..124 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 252..271 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 471..987 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" misc_feature 1051..1392 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 1459..1875 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1805..1827 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1843..1859 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" primer_bind complement(2088..2105) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2315..2334 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2496..3215 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2541..2562) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2802..2821) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3149..3170 /label=EGFP-C /note="EGFP, forward primer" LTR 3377..3891 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 4053..4069 /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(4077..4107) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4122..4143 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4260..4277) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4431..5019) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4511..4530) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5190..6050) /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 5813..5832 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6051..6155) /gene="bla" /label=AmpR promoter primer_bind 6223..6241 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6279..6301) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6401..6420 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg 61 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa 121 aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg 181 gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga 241 tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg 301 atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata 361 tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac 421 gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac 481 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata 541 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg 601 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg 661 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg 721 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg 781 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg 841 gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct 901 tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct cagattgatt 961 gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga gacccctgcc 1021 cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc 1081 tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct gtactagtta 1141 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc 1201 cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga 1261 agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac 1321 ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg 1381 tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg 1441 tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac 1501 tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg 1561 gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc 1621 tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga 1681 cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc 1741 tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct 1801 ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact 1861 ccttctctag gcgccggaat tcagatctta cgtagctagc ggatcccaat tgctcgagcg 1921 ggatcaattc cgcccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg 1981 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg 2041 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg 2101 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca 2161 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct 2221 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca 2281 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa 2341 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg 2401 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg 2461 ggacgtggtt ttcctttgaa aaacacgata ataccatggt gagcaagggc gaggagctgt 2521 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca 2581 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct 2641 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc 2701 tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca 2761 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga 2821 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca 2881 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc 2941 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc 3001 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca 3061 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga 3121 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg 3181 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct agagtcgacc 3241 tgcaggcatg caagcttcag gtagccggct aacgttaaca accggtacct ctagaactat 3301 agctagcatg cgcaaattta aagcgctgat atcgataaaa taaaagattt tatttagtct 3361 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag cttaagtaac 3421 gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag atcaaggtta 3481 ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc 3541 ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag cagtttctag 3601 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg 3661 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat 3721 aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc gtcgcccggg 3781 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct 3841 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tgggtaacag 3901 tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag taatcctgac 3961 tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt tcattatgga 4021 cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat tccacacaac 4081 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 4141 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 4201 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 4261 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 4321 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 4381 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 4441 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 4501 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 4561 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 4621 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 4681 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 4741 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 4801 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 4861 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 4921 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 4981 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 5041 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 5101 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 5161 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 5221 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 5281 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 5341 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 5401 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 5461 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 5521 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 5581 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 5641 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 5701 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 5761 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 5821 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 5881 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 5941 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 6001 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 6061 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 6121 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 6181 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 6241 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg 6301 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg 6361 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta 6421 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 6481 atcaggcgcc attcgccatt caggctgcgc aa //