LOCUS Exported 7336 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of cytosolic Grx1-roGFP2 (retroviral vector). ACCESSION . VERSION . KEYWORDS pLPCX cyto Grx1-roGFP2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7336) AUTHORS Gutscher M, Pauleau AL, Marty L, Brach T, Wabnitz GH, Samstag Y, Meyer AJ, Dick TP TITLE Real-time imaging of the intracellular glutathione redox potential. JOURNAL Nat Methods. 2008 Jun;5(6):553-9. doi: 10.1038/nmeth.1212. Epub 2008 May 11. PUBMED 18469822 REFERENCE 2 (bases 1 to 7336) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7336 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 414..589 /label=5' LTR (truncated) /note="truncated long terminal repeat from Moloney murine sarcoma virus" misc_feature 652..851 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 1052..1468 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1398..1420 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1436..1452 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1565..2164 /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(1565..1584) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2061..2081 /label=Puro-F /note="Puromycin resistance gene, forward primer" enhancer 2339..2642 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2643..2846 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2797..2817 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 3313..4029 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(3355..3376) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3616..3635) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3963..3984 /label=EGFP-C /note="EGFP, forward primer" LTR 4075..4668 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(4771..4793) /label=pGEX 3' /note="pGEX vectors, reverse primer" protein_bind complement(4921..4954) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(5014..5047) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind 5067..5086 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5167..5184) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5338..5926) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5418..5437) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6097..6957) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6720..6739 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6958..7062) /gene="bla" /label=AmpR promoter primer_bind 7130..7148 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat 61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc 121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca 181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg 241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa 301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac 361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa 421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac 481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg 541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt 601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc 661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg 721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt 781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg 841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt 901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa 961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct 1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt 1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa 1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc 1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc 1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt 1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc 1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta 1441 tccagccctc actccttctc taggcgccgg aatgatcccc cgggctgcag gtcggccgcc 1501 acgaccggtg ccgccaccat cccctgaccc acgcccctga cccctcacaa ggagacgacc 1561 ttccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc 1621 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc 1681 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct 1741 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc 1801 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag 1861 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa 1921 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct 1981 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt 2041 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac 2101 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc 2161 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc 2221 cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgac 2281 tctagtaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 2341 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 2401 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 2461 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 2521 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 2581 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 2641 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 2701 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 2761 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 2821 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc 2881 ggactcagat ctatggctca agagtttgtg aactgcaaaa tccagcctgg gaaggtggtt 2941 gtgttcatca agcccacctg cccgtactgc aggagggccc aagagatcct cagtcaattg 3001 cccatcaaac aagggcttct ggaatttgtc gatatcacag ccaccaacca cactaacgag 3061 attcaagatt atttgcaaca gctcacggga gcaagaacgg tgcctcgagt ctttattggt 3121 aaagattgta taggcggatg cagtgatcta gtctctttgc aacagagtgg ggaactgctg 3181 acgcggctaa agcagattgg agctctgcag actagtggtg gttcaggtgg tggtggttca 3241 ggtggtggtg gttcaggtgg aggaggatca ggaggaggag gatcaggagg aggaggatca 3301 ggaggagaat tcgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc 3361 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat 3421 gccacctacg gcaagctgac cctgaagttc atctccacca ccggcaagct gcccgtgccc 3481 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac 3541 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc 3601 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc 3661 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc 3721 ctggggcaca agctggagta caactacaac tgccacaacg tctatatcat ggccgacaag 3781 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg 3841 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc 3901 gacaaccact acctgagcac ctgctccgcc ctgagcaaag accccaacga gaagcgcgat 3961 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg 4021 tacaagtaaa tcgataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa 4081 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa 4141 aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagctga 4201 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac 4261 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg 4321 ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa 4381 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact 4441 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa 4501 gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc 4561 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg 4621 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc 4681 cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt aagctggctg 4741 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 4801 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 4861 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtactg 4921 ataacttcgt ataatgtatg ctatacgaag ttattaggtc tgaagaggag tttacgtcca 4981 gccaagctag ggccgcgatc cggaaccctt aatataactt cgtataatgt atgctatacg 5041 aagttatcag tactggctta actatgcggc atcagagcag attgtactga gagtgcacca 5101 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc 5161 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 5221 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 5281 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 5341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 5401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 5461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 5521 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 5581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 5641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 5701 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 5761 ctacggctac actagaagra cagtatttgg tatctgcgct ctgctgaagc cagttacctt 5821 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 5881 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 5941 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 6001 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 6061 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc 6121 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta 6181 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga 6241 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg 6301 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc 6361 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat 6421 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag 6481 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat 6541 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa 6601 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa 6661 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga 6721 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg 6781 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc 6841 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg 6901 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact 6961 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat 7021 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt 7081 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat 7141 cacgaggccc tttcgtcttc aagaattgct agcaattgct agcaattgct agcaattcat 7201 accagatcac cgaaaactgt cctccaaatg tgtccccctc acactcccaa attcgcgggc 7261 ttctgcctct tagaccactc taccctattc cccacactca ccggagccaa agccgcggcc 7321 cttccgtttc tttgct //