LOCUS Exported 8018 bp ds-DNA circular SYN 08-JAN-2024 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBABE-puro-GFP-progerin SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8018) AUTHORS Scaffidi P, Misteli T TITLE Lamin A-dependent misregulation of adult stem cells associated with accelerated ageing. JOURNAL Nat Cell Biol. 2008 Mar 2. ():. PUBMED 18311132 REFERENCE 2 (bases 1 to 8018) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 8, 2024 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8018 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 17..486 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 551..750 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 951..1367 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1297..1319 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1335..1351 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" regulatory 1406..1415 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1412..2128 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1412..2128 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1457..1478) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1718..1737) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2065..2086 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(4357..4377) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4362..4691 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4542..4677 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4604..4623 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4701..5300 /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(4701..4720) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5197..5217 /label=Puro-F /note="Puromycin resistance gene, forward primer" LTR 5521..5990 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(6120..6137) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6291..6879) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6371..6390) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7039..7899) /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 7662..7681 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7900..8004) /gene="bla" /label=AmpR promoter ORIGIN 1 aagtgccacc tgcagcctga atatgggcca aacaggatat ctgtggtaag cagttcctgc 61 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg 121 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc 181 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga 241 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct 301 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt 361 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg 421 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt 481 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc 541 accgggaggc aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga 601 ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc 661 gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact 721 ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt 781 caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat 841 ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg 901 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg 961 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca 1021 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa 1081 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta 1141 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga 1201 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc 1261 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc 1321 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccggc cggatcccag 1381 tgtggtggta cgtacgctac cggtcgccac catggtgagc aagggcgagg agctgttcac 1441 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt 1501 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac 1561 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca 1621 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc 1681 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg 1741 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga 1801 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa 1861 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca 1921 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg 1981 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa 2041 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat 2101 cactctcggc atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttcgaa 2161 ttctgcagtc gacggtaccg cgggcagcgc tgccaacctg ccggccatgg agaccccgtc 2221 ccagcggcgc gccacccgca gcggggcgca ggccagctcc actccgctgt cgcccacccg 2281 catcacccgg ctgcaggaga aggaggacct gcaggagctc aatgatcgct tggcggtcta 2341 catcgaccgt gtgcgctcgc tggaaacgga gaacgcaggg ctgcgccttc gcatcaccga 2401 gtctgaagag gtggtcagcc gcgaggtgtc cggcatcaag gccgcctacg aggccgagct 2461 cggggatgcc cgcaagaccc ttgactcagt agccaaggag cgcgcccgcc tgcagctgga 2521 gctgagcaaa gtgcgtgagg agtttaagga gctgaaagcg cgcaatacca agaaggaggg 2581 tgacctgata gctgctcagg ctcggctgaa ggacctggag gctctgctga actccaagga 2641 ggccgcactg agcactgctc tcagtgagaa gcgcacgctg gagggcgagc tgcatgatct 2701 gcggggccag gtggccaagc ttgaggcagc cctaggtgag gccaagaagc aacttcagga 2761 tgagatgctg cggcgggtgg atgctgagaa caggctgcag accatgaagg aggaactgga 2821 cttccagaag aacatctaca gtgaggagct gcgtgagacc aagcgccgtc atgagacccg 2881 actggtggag attgacaatg ggaagcagcg tgagtttgag agccggctgg cggatgcgct 2941 gcaggaactg cgggcccagc atgaggacca ggtggagcag tataagaagg agctggagaa 3001 gacttattct gccaagctgg acaatgccag gcagtctgct gagaggaaca gcaacctggt 3061 gggggctgcc cacgaggagc tgcagcagtc gcgcatccgc atcgacagcc tctctgccca 3121 gctcagccag ctccagaagc agctggcagc caaggaggcg aagcttcgag acctggagga 3181 ctcactggcc cgtgagcggg acaccagccg gcggctgctg gcggaaaagg agcgggagat 3241 ggccgagatg cgggcaagga tgcagcagca gctggacgag taccaggagc ttctggacat 3301 caagctggcc ctggacatgg agatccacgc ctaccgcaag ctcttggagg gcgaggagga 3361 gaggctacgc ctgtccccca gccctacctc gcagcgcagc cgtggccgtg cttcctctca 3421 ctcatcccag acacagggtg ggggcagcgt caccaaaaag cgcaaactgg agtccactga 3481 gagccgcagc agcttctcac agcacgcacg cactagcggg cgcgtggccg tggaggaggt 3541 ggatgaggag ggcaagtttg tccggctgcg caacaagtcc aatgaggacc agtccatggg 3601 caattggcag atcaagcgcc agaatggaga tgatcccttg ctgacttacc ggttcccacc 3661 aaagttcacc ctgaaggctg ggcaggtggt gacgatctgg gctgcaggag ctggggccac 3721 ccacagcccc cctaccgacc tggtgtggaa ggcacagaac acctggggct gcgggaacag 3781 cctgcgtacg gctctcatca actccactgg ggaagaagtg gccatgcgca agctggtgcg 3841 ctcagtgact gtggttgagg acgacgagga tgaggatgga gatgacctgc tccatcacca 3901 ccacggctcc cactgcagca gctcggggga ccccgctgag tacaacctgc gctcgcgcac 3961 cgtgctgtgc gggacctgcg ggcagcctgc cgacaaggca tctgccagcg gctcaggagc 4021 ccagagcccc cagaactgca gcatcatgta atctgggacc tgccaggcag gggtgggggt 4081 ggaggcttcc tgcgtcctcc tcacctcatg cccaccccct gccctgcacg tcatgggagg 4141 gggcttgaag ccaaagaaaa ataacccttt ggtttttttc ttctgtattt ttttttctaa 4201 gagaagttat tttctacagt ggttttatac tgaaggaaaa acacaagcaa aaaaaaaaaa 4261 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa gcgtctagaa accctttaga 4321 tctgctgcag cgcgccagca cagtggtcga ccctgtggaa tgtgtgtcag ttagggtgtg 4381 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 4441 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 4501 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 4561 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 4621 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 4681 gcttttgcaa aaagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg 4741 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc 4801 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc 4861 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg 4921 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc 4981 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg 5041 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc 5101 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg 5161 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc 5221 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga 5281 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg 5341 cacgacccca tgcatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa 5401 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat 5461 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag 5521 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa 5581 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc 5641 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag 5701 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg 5761 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat 5821 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg 5881 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct 5941 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat 6001 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat 6061 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 6121 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 6181 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 6241 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 6301 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 6361 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 6421 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 6481 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 6541 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 6601 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 6661 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 6721 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 6781 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 6841 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 6901 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 6961 tcaaaaagga tcttcaccta gatccttttg cggccgcaaa tcaatctaaa gtatatatga 7021 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 7081 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 7141 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 7201 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 7261 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 7321 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 7381 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 7441 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 7501 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 7561 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 7621 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 7681 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 7741 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 7801 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 7861 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 7921 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 7981 aaataaacaa ataggggttc cgcgcacatt tccccgaa //