LOCUS Exported 9349 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Generation of retrovirus for the overexpression of Nr4a1 mutant (zinc finger 2 deletion). ACCESSION . VERSION . KEYWORDS MSCV-HA-Nr4a1-Zf2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9349) AUTHORS Zhang Y, Federation AJ, Kim S, O'Keefe JP, Lun M, Xiang D, Brown JD, Steinhauser ML TITLE Targeting nuclear receptor NR4A1-dependent adipocyte progenitor quiescence promotes metabolic adaptation to obesity. JOURNAL J Clin Invest. 2018 Nov 1;128(11):4898-4911. doi: 10.1172/JCI98353. Epub 2018 Oct 2. PUBMED 30277475 REFERENCE 2 (bases 1 to 9349) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9349 /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" CDS 1431..1457 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(3231..3254) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 3592..3611 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 3680..4279 /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(3680..3699) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4176..4196 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 4357..4909 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4501..4518) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 4728..4747 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4921..5640 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4966..4987) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5227..5246) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5574..5595 /label=EGFP-C /note="EGFP, forward primer" LTR 5742..6256 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 6418..6434 /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(6442..6472) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6487..6508 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6625..6642) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6796..7384) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6876..6895) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7555..8415) /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 8178..8197 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8416..8520) /gene="bla" /label=AmpR promoter primer_bind 8588..8606 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(8644..8666) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 8766..8785 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8979..9001 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 9129..9148 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 9348..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 tggaacctaa 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 gatctctcga ggccaccatg tacccatacg 1441 atgttccgga ttacgctccc tgtattcaag ctcaatatgg aacaccagca acgagcccag 1501 gaccgcgtga ccacctgacc ggtgatcccc tggcccttga gttcggcaag cctaccatgg 1561 acctggccag ccccgagaca gcacctgccg cacctgctac actgcccagc ttcagcacct 1621 tcatggacgg gtacaccgga gagtttgaca ccttcctcta ccagctgccg gggacgaccc 1681 agccgtgctc ctcagcttgt tcctctgcct cctccacgtc ttcttcctca tcctcggcca 1741 cctcccccgc ttcggcgtcc ttcaagtttg aggacttcca ggtgtacggc tgctacccgg 1801 gcaccctgag cggcccatta gatgagaccc tatcctccag cggctctgag tactatggca 1861 gtccctgctc agccccctcg ccatctacac ccaacttcca gccgtcccag ctttctccct 1921 gggacggctc atttggccac ttctccccga gccagactta tgaaggcctc tgggcatgga 1981 cagagcagtt gcctaaggct tcttcagggc ctccgccacc tccaaccttc ttctccttca 2041 gtcctcccac tggccccagc cccagcctgg cccagagttc tctgaaattg ttcccaccac 2101 cagccaccca ccagcttggg gagggggaga gctattccat gccagcagct ttccccggct 2161 tggcacccac ctctccgaac cgtgacactt ccggcattct ggacgcaccc gtgacctcca 2221 ccaagtcccg gagcggggct tcaggtggca gcgagggccg ctgtgcagtc tgtggtgaca 2281 atgcttcgtg tcagcactat ggggtccgca cctgtgaggg ctgcaagggc ttcttcaagc 2341 gcacagtaca gaaaagcgcc aagtacatcc tggctgtggg catggtgaag gaagttgtac 2401 ggacagacag cctaaaaggg cggcggggcc ggctaccttc aaaacccaag cagcctccag 2461 atgcctcccc taccaatctt ctcacttccc tcatccgggc acacttggac tccgggccta 2521 gcactgccaa attggactat tccaagttcc aggaactggt gctgccccgc ttcgggaagg 2581 aagatgccgg tgacgtgcaa caattttatg acttgctctc tggttccctg gacgttatcc 2641 gaaagtgggc agaaaaaatc cctggcttca ttgagctttg cccaggagac caagacctgt 2701 tgctagagtc tgccttcctg gaactcttca tcctccgcct ggcataccga tctaaacccg 2761 gtgaggggaa gctcatcttc tgctcaggcc tggtactaca ccagctgcag tgtgcccgtg 2821 gctttggtga ttggattgac aacatcctgg ccttctcacg gtccctgcac agcttgggtg 2881 ttgatgttcc cgcctttgcc tgcctgtccg ctctggtcct catcactgat cgacacgggc 2941 tccaggaccc tcgtcgggtg gaagagctgc agaatcgcat tgctagctgt ctgaaggagc 3001 acatggctac cgtggcagga gacccacagc cggccagctg cctgtcacgt ctgctgggca 3061 aactgcctga gcttcggacc ctgtgcactc aaggcctgca gcgcatcttt tgcctcaagt 3121 tggaggactt ggtaccccct ccacctattg tggacaagat ctttatggac acattgtctt 3181 tctgagaatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt 3241 tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca 3301 catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta 3361 ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac 3421 aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga 3481 agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc 3541 agaggctggg aaggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca 3601 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga 3661 cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga 3721 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg 3781 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct 3841 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc 3901 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg 3961 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc 4021 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca 4081 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc 4141 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct 4201 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac 4261 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc 4321 acgaccccat gcatccaatt ccgccccccc cccctaacgt tactggccga agccgcttgg 4381 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca 4441 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc 4501 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag 4561 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg 4621 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac 4681 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa 4741 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc 4801 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc 4861 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc 4921 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 4981 ggcgacgtaa acggccacaa gttcagcgtg tctggcgagg gcgagggcga tgccacctac 5041 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 5101 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 5161 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 5221 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 5281 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 5341 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 5401 ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 5461 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 5521 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 5581 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa 5641 tgaattaatt aagaattatc aagcttatcg ataccgtcga cctgcagcca agcttatcga 5701 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta 5761 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag 5821 aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg 5881 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg 5941 cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac 6001 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg 6061 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt 6121 cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca 6181 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc 6241 agcgggggtc tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag 6301 gagtcgaata ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa 6361 tactcctgaa atagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg 6421 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg 6481 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc 6541 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 6601 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 6661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 6721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 6781 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 6841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 6901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 6961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 7021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 7081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 7141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 7201 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 7261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 7321 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 7381 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 7441 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 7501 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 7561 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 7621 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 7681 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 7741 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 7801 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 7861 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 7921 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 7981 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 8041 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 8101 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 8161 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 8221 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 8281 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 8341 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 8401 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 8461 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 8521 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac 8581 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga 8641 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg 8701 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa 8761 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca 8821 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg 8881 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg 8941 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac 9001 gacggcgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg 9061 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct 9121 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg 9181 ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca 9241 gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag 9301 ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaa //