LOCUS Exported 8100 bp ds-DNA circular SYN 11-FEB-2019 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8100) AUTHORS . TITLE Direct Submission JOURNAL Exported Friday, Jan 8, 2021 from SnapGene Viewer 5.2.4 https://www.snapgene.com FEATURES Location/Qualifiers source 1..8100 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(455..1315) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 455 .. 1246 / #ccffcc 2: 1247 .. 1315 / #ccffcc / signal sequence Cleavage site after base 1246" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(1316..1420) /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: LEFT" LTR 2248..2764 /label=5' LTR /note="5' long terminal repeat from murine embryonic stem cell virus" /note="color: #ffe4c4" misc_feature 2828..3169 /label=MESV Psi /note="packaging signal of murine embryonic stem cell virus" /note="color: #ffe4c4" CDS 3236..3652 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" /note="color: #993366; direction: RIGHT" regulatory 3665..3674 /regulatory_class="other" /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /note="color: #a6acb3" CDS 3674..3739 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /note="color: #cc99b2 Cleavage site after base 3739" /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 3740..5329 /codon_start=1 /label=Tox1 /note="color: #993366" /translation="LAAAMDVRFYPPPAQPAAAPAAPCLGPSPCLDPYYCNKFDGENMY MSMTEPSQDYVPASQSYPGPSLESEDFNIPPITPPSLPDHSLVHLNEVESGYHSLCHPM NHNGLLPFHPQTMDLPEITVSNMLGQDGALLSNSISVMQEIGNAEGAQYSSHPQMAAMR PRGQPTDIRQQASMMQPGQLTTINQSQLSAQLGLNMGGTNVAHNSPSPPGSKSATPSPS SSVHEDECEDASKINGGEKRPASDMGKKPKTPKKKKKKDPNEPQKPVSAYALFFRDTQA AIKGQNPNATFGEVSKIVASMWDGLGEEQKQVYKKKTEAAKKEYLKQLAAYRASLVSKS YTDPVDVKTSQPPQLVNSKPSVFHGPSQAHSALYLSSHYHQQPGMTPQLTAMHPSLPRN IAPKPNNQMPVTVSIANMAVSPPPPLQISPPLHQHLSMQQHQSLAMQQPLGSQLPMQVQ TALHSPTMQQGFTLQPDYQTIINPTSTAAQVVTQAMEYVRSGCRNPPPQPVDWSTDYCS SGGMQRDKALYLT" misc_feature 5357..5909 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" /note="color: #a6acb3" CDS 5921..6640 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /note="This feature has 3 segments: 1: 5921 .. 5923 / #05fd14 2: 5924 .. 5926 / #05fd14 / 1a 3: 5927 .. 6640 / #05fd14" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" LTR 6742..7256 /label=3' LTR /note="3' long terminal repeat from murine embryonic stem cell virus" /note="color: #ffe4c4" protein_bind 7418..7434 /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)." /note="color: #31849b" promoter complement(7442..7472) /label=lac promoter /note="promoter for the E. coli lac operon" /note="This reverse directional feature has 3 segments: 1: 7442 .. 7448 / #ffffff / -10 2: 7449 .. 7466 / #ffffff 3: 7467 .. 7472 / #ffffff / -35" protein_bind 7487..7508 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /note="color: #31849b" rep_origin complement(join(7796..8100,1..284)) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" ORIGIN 1 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 61 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 121 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 1021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 1081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 1141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 1201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 1261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 1321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 1381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 1441 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca 1501 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc 1561 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg 1621 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga 1681 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat 1741 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc 1801 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt 1861 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc 1921 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc 1981 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc 2041 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc 2101 gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt 2161 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat 2221 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct 2281 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc 2341 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa 2401 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc 2461 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct 2521 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc 2581 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact 2641 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg 2701 actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt 2761 catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc 2821 gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc 2881 cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg 2941 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag 3001 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac 3061 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc 3121 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag 3181 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca 3241 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc 3301 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg 3361 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca 3421 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat 3481 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc 3541 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc 3601 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattag 3661 atctgccacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta 3721 caaggatgac gatgacaagc ttgcggccgc gatggacgta agattttatc ctcctccagc 3781 ccagcccgct gctgcgcccg ccgctccttg tctgggacct tctccctgcc tggaccccta 3841 ctattgcaac aagtttgacg gggagaacat gtatatgagc atgacagagc cgagccagga 3901 ctatgtgcca gccagccagt cctaccctgg accaagtctg gaaagtgaag attttaacat 3961 cccacccatc acgcctccct ccctccctga ccactctctg gtgcacctga atgaggtaga 4021 atccggttac cactctctgt gtcaccccat gaatcacaat ggcctgctcc cgttccatcc 4081 acaaaccatg gacctgccag agatcacggt gtccaacatg ctgggccaag atggcgcact 4141 gctctccaat tccatctctg tgatgcaaga gattgggaac gcagaaggag ctcagtacag 4201 ctcccatcct cagatggcag ccatgaggcc gagaggccag cctacagaca tcagacagca 4261 ggcaagcatg atgcagcccg gccagctgac caccatcaac cagtcgcagc tgagcgcaca 4321 gttgggcttg aacatgggag gaaccaatgt ggcccacaac tccccttctc cacccgggag 4381 caagtctgca accccttccc catccagctc agtgcatgag gatgagtgtg aggatgcctc 4441 caagatcaat ggaggagaga agcggcctgc ctctgatatg gggaaaaaac ccaaaactcc 4501 caagaagaag aagaagaagg atcctaatga gccccagaag cctgtgtctg cctatgcttt 4561 gttctttcgt gatacccagg ctgccattaa gggccagaat ccaaatgcta cttttggtga 4621 agtatcgaag attgtggcat caatgtggga tggcctagga gaagaacaga aacaggtcta 4681 taagaagaaa accgaggctg caaagaagga atacctgaag cagctagcag catacagagc 4741 cagccttgta tccaagagct acactgatcc tgttgatgtt aagacatctc agccacccca 4801 gctggtcaac tcaaagccgt cagtattcca tgggcccagc caggcccact cagctctgta 4861 cctaagttct cactatcacc aacaaccagg aatgactcct cagctcaccg ccatgcatcc 4921 gagcctcccc agaaacatag cacctaagcc caataaccaa atgccagtga ctgtctccat 4981 agccaacatg gctgtgtccc caccaccgcc ccttcagatc agcccacctc ttcatcagca 5041 tctcagcatg cagcagcatc agtcgcttgc catgcagcag ccccttggga gtcagctccc 5101 catgcaggtc cagactgcct tacactcgcc caccatgcag caaggattta ctcttcaacc 5161 cgactatcag actattatca accctacatc tacagctgca caagttgtca cccaagcgat 5221 ggagtacgtg cgttctgggt gcagaaatcc gcccccacag cccgtggact ggagtaccga 5281 ctactgcagt agtgggggca tgcagaggga caaagcgctg tatctcacct gataagaatt 5341 ccgccccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt 5401 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct 5461 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa 5521 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg 5581 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc 5641 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg 5701 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg 5761 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc 5821 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg 5881 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga 5941 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa 6001 gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt 6061 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta 6121 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc 6181 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta 6241 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa 6301 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa 6361 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa 6421 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac 6481 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc 6541 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc 6601 cgccgggatc actctcggca tggacgagct gtacaagtaa tgaattaatt aagaattatc 6661 aagcttatcg ataccgtcga cctgcagcca agcttatcga taaaataaaa gattttattt 6721 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa 6781 gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa 6841 ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc 6901 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt 6961 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt 7021 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc 7081 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc 7141 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg 7201 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatgggt 7261 aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata ttaagtaatc 7321 ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa atagttcatt 7381 atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc acaattccac 7441 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 7501 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 7561 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 7621 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 7681 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 7741 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 7801 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 7861 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 7921 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 7981 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 8041 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc //