LOCUS Exported 8431 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS MSCV h c-MYC IRES GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8431) TITLE Cleveland lab plasmids REFERENCE 2 (bases 1 to 8431) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8431 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 580..921 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" CDS 988..1404 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1334..1356 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1372..1388 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1649..2968 /codon_start=1 /product="human c-Myc proto-oncogene" /label=c-Myc /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA" primer_bind complement(3451..3468) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3678..3697 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 3858..4577 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 3858..4574 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3903..3924) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4164..4183) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4511..4532 /label=EGFP-C /note="EGFP, forward primer" LTR 4739..5254 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" protein_bind 5416..5432 /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(5440..5470) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5485..5506 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5623..5640) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5794..6382) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5874..5893) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6553..7413) /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 7176..7195 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7414..7518) /gene="bla" /label=AmpR promoter primer_bind 7586..7604 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7642..7664) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 7764..7783 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 7977..7999 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7991..8008 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7992..8008 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 8211..8230 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" LTR 8430..516 /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 gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt 301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc 361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat 421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc 481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag 541 acccctgcct agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc 601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg 661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga 721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg 781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga 841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga 901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact 961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc 1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag 1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga 1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc 1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac 1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc 1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca 1381 gccctcactc cttctctagg cgccggaatt cggccccccc ccccttgccg catccacgaa 1441 actttgccca tagcagcggg cgggcacttt gcactggaac ttacaacacc cgagcaagga 1501 cgcgactctc ccgacgcggg gaggctattc tgcccatttg gggacacttc cccgccgctg 1561 ccaggacccg cttctctgaa aggctctcct tgcagctgct tagacgctgg atttttttcg 1621 ggtagtggaa aaccagcctc ccgcgacgat gcccctcaac gttagcttca ccaacaggaa 1681 ctatgacctc gactacgact cggtgcagcc gtatttctac tgcgacgagg aggagaactt 1741 ctaccagcag cagcagcaga gcgagctgca gcccccggcg cccagcgagg atatctggaa 1801 gaaattcgag ctgctgccca ccccgcccct gtcccctagc cgccgctccg ggctctgctc 1861 gccctcctac gttgcggtca cacccttctc ccttcgggga gacaacgacg gcggtggcgg 1921 gagcttctcc acggccgacc agctggagat ggtgaccgag ctgctgggag gagacatggt 1981 gaaccagagt ttcatctgcg acccggacga cgagaccttc atcaaaaaca tcatcatcca 2041 ggactgtatg tggagcggct tctcggccgc cgccaagctc gtctcagaga agctggcctc 2101 ctaccaggct gcgcgcaaag acagcggcag cccgaacccc gcccgcggcc acagcgtctg 2161 ctccacctcc agcttgtacc tgcaggatct gagcgccgcc gcctcagagt gcatcgaccc 2221 ctcggtggtc ttcccctacc ctctcaacga cagcagctcg cccaagtcct gcgcctcgca 2281 agactccagc gccttctctc cgtcctcgga ttctctgctc tcctcgacgg agtcctcccc 2341 gcagggcagc cccgagcccc tggtgctcca tgaggagaca ccgcccacca ccagcagcga 2401 ctctgaggag gaacaagaag atgaggaaga aatcgatgtt gtttctgtgg aaaagaggca 2461 ggctcctggc aaaaggtcag agtctggatc accttctgct ggaggccaca gcaaacctcc 2521 tcacagccca ctggtcctca agaggtgcca cgtctccaca catcagcaca actacgcagc 2581 gcctccctcc actcggaagg actatcctgc tgccaagagg gtcaagttgg acagtgtcag 2641 agtcctgaga cagatcagca acaaccgaaa atgcaccagc cccaggtcct cggacaccga 2701 ggagaatgtc aagaggcgaa cacacaacgt cttggagcgc cagaggagga acgagctaaa 2761 acggagcttt tttgccctgc gtgaccagat cccggagttg gaaaacaatg aaaaggcccc 2821 caaggtagtt atccttaaaa aagccacagc atacatcctg tccgtccaag cagaggagca 2881 aaagctcatt tctgaagagg acttgttgcg gaaacgacga gaacagttga aacacaaact 2941 tgaacagcta cggaactctt gtgcgtaagg aaaagtaagg aaaatgattc cttctaacag 3001 aaatgtcctg agcaatcacc tatgaacttg tttcaaatgc atgatcaaat gcaacctcac 3061 aaccttggct gagtcttgag actgaaagat ttagccataa tgtaaactgc ctcaaattgg 3121 actttgggca taaaagaact tttttatgct taccatcttt tttttttctt taacagattt 3181 gtatttaaga attgttttta aaaaatttta agatttacac aatgtttctc tgtaaatatt 3241 gccattaaat gtaaataact tccgaattcg ttaacctcga gcgggatcaa ttccgccccc 3301 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt 3361 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct 3421 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga 3481 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga 3541 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac 3601 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag 3661 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc 3721 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg 3781 tttagtcgag gttaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg 3841 aaaaacacga taataccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca 3901 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg 3961 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc 4021 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct 4081 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc 4141 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt 4201 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg 4261 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg 4321 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg 4381 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 4441 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga 4501 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg 4561 acgagctgta caagtaaagc ggccgcgact ctagagtcga cctgcaggca tgcaagcttc 4621 aggtagccgg ctaacgttaa caaccggtac ctctagaact atagctagca tgcgcaaatt 4681 taaagcgctg atatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat 4741 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg 4801 gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga gagacagcag 4861 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa 4921 cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat cagatgtttc 4981 cagggtgccc caaggacctg aaaatgaccc tgtgccttat ttgaactaac caatcagttc 5041 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc 5101 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt gtatccaata 5161 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg 5221 agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg aagttggaga 5281 acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta gccactgttt 5341 tgaatccaca tactccaata ctccgtaaat agttcattat ggacagcgca gaaagagctg 5401 gggagaattg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 5461 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 5521 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 5581 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 5641 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 5701 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 5761 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 5821 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 5881 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 5941 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 6001 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 6061 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 6121 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 6181 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 6241 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 6301 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 6361 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 6421 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 6481 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 6541 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 6601 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 6661 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 6721 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 6781 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 6841 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 6901 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 6961 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 7021 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 7081 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 7141 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 7201 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 7261 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 7321 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 7381 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 7441 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 7501 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 7561 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg 7621 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg 7681 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg 7741 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat 7801 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc 7861 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca 7921 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca 7981 gtcacgacgt tgtaaaacga cggccagtgc cannnncgct ctcccttatg cgactcctgc 8041 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt 8101 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg 8161 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg 8221 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg 8281 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt 8341 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag 8401 attttattta gtctccagaa aaaaaaaaaa a //