LOCUS Exported 6579 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Ccnb2 shRNA in pMKO.1 retroviral vector. ACCESSION . VERSION . KEYWORDS pMKO.1-shCcnb2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6579) 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 6579) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6579 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..241 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 1..21 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 172..191 /label=LKO.1 5' /note="Human U6 promoter, forward primer" promoter 339..668 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 519..654 /label=SV40 ori /note="SV40 origin of replication" CDS 678..1277 /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(678..697) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1174..1194 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 1473..1898 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from Moloney murine leukemia virus" primer_bind complement(2001..2023) /label=pGEX 3' /note="pGEX vectors, reverse primer" promoter 2164..2493 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2344..2479 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2515..2534 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(2615..2632) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2786..3374) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2866..2885) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3545..4405) /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 4168..4187 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4406..4510) /gene="bla" /label=AmpR promoter primer_bind 4788..4806 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" enhancer 4906..5285 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 5286..5489 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5440..5460 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 5490..5665 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 5728..5927 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 6128..6544 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 6474..6496 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 6512..6528 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga 241 cgaaacaccg ggatgttgaa cagcacactt tactcgagta aagtgtgctg ttcaacatct 301 ttttgaattc gccagcacag tggtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa 361 agtccccagg cttcccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 421 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 481 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 541 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 601 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 661 tttgcaaaaa gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg 721 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc 781 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca 841 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg 901 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca 961 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc 1021 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc 1081 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg 1141 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg 1201 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc 1261 gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac 1321 gaccccatgc atcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta 1381 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa 1441 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc 1501 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag 1561 aagttcagat caaggtcagg aacagatgga acagggtcga ccctagagaa ccatcagatg 1621 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag 1681 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa 1741 cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca 1801 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct 1861 ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg 1921 agacccctgc ccagggacca ccgacccacc accgggaggt aagctggctg cctcgcgcgt 1981 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt 2041 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg 2101 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtagat ccggctgtgg 2161 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 2221 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 2281 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 2341 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 2401 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga 2461 ggaggctttt ttggaggcct aggcttttgc aaaaagctta ctggcttaac tatgcggcat 2521 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa 2581 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 2641 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 2701 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 2761 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 2821 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 2881 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 2941 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc 3001 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 3061 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 3121 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 3181 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta 3241 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 3301 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 3361 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 3421 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 3481 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 3541 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 3601 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 3661 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 3721 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 3781 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 3841 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 3901 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 3961 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 4021 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 4081 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 4141 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 4201 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 4261 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 4321 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 4381 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 4441 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 4501 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt 4561 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg 4621 caaaatccct taattattga agcatttatc agggttattg tctcatgagc ggatacatat 4681 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 4741 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca 4801 cgaggccctt tcgtcttcaa gaattagctt ggccattgca tacgttgtat ccatatcata 4861 atatgtacat ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga 4921 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc 4981 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 5041 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 5101 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 5161 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 5221 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 5281 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg 5341 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac 5401 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg 5461 tacggtggga ggtctatata agcagagctc aataaaagag cccacaaccc ctcactcggc 5521 gcgccagtct tccgatagac tgcgtcgccc gggtacccgt attcccaata aagcctcttg 5581 ctgtttgcat ccgaatcgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga 5641 ctacccacga cgggggtctt tcatttgggg gctcgtccgg gatttggaga cccctgccca 5701 gggaccaccg acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga 5761 ttgtctagtg tctatgtttg atgttatgcg cctgcgtctg tactagttag ctaactagct 5821 ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg 5881 gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat 5941 gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt 6001 cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct 6061 gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga 6121 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt 6181 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg 6241 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga 6301 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca 6361 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc 6421 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga 6481 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg 6541 cgccggaatt gaagatctgt cgacaaggtc gggcaggaa //