LOCUS Exported 4791 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: Protein Expression Vector , Excitation: 488, Emission: 507. ACCESSION . VERSION . KEYWORDS mEGFP-pBAD SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4791) TITLE Michael Davidson Empty Backbones REFERENCE 2 (bases 1 to 4791) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4791 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 117..134 /label=L4440 /note="L4440 vector, forward primer" misc_feature 149..289 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(215..234) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 375..397 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(553..1431) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 1458..1742 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 1665..1684 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" CDS 1788..1805 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 1809..1841 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 1845..1868 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 1884..2603 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1929..1950) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2190..2209) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2537..2558 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2649..2666) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 2819..2905 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2997..3024 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 3043..3134 /gene="bla" /label=AmpR promoter CDS 3135..3995 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3353..3372) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4166..4754 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4655..4674 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc 61 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg 121 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt 181 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt 241 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc 301 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca 361 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg 421 cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca 481 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg 541 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc 601 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg 661 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag 721 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg 781 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc 841 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg 901 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg 961 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat 1021 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc 1081 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa 1141 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc 1201 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt 1261 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg 1321 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg 1381 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat 1441 actcccgcca ttcagagaag aaaccaattg tccatattgc atcagacatt gccgtcactg 1501 cgtcttttac tggctcttct cgctaaccaa accggtaacc ccgcttatta aaagcattct 1561 gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg 1621 cagaaaagtc cacattgatt atttgcacgg cgtcacactt tgctatgcca tagcattttt 1681 atccataaga ttagcggatc ctacctgacg ctttttatcg caactctcta ctgtttctcc 1741 atacccgttt tttgggctaa caggaggaat taaccatggg gggttctcat catcatcatc 1801 atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg tacgacgatg 1861 acgataagga tccgagctcg agcatggtga gcaagggcga ggagctgttc accggggtgg 1921 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 1981 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca 2041 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca 2101 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 2161 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 2221 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 2281 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 2341 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg 2401 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 2461 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca 2521 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg 2581 gcatggacga gctgtacaag taagaattcg aagcttggct gttttggcgg atgagagaag 2641 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg 2701 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc 2761 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca 2821 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgccggt 2881 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg 2941 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa 3001 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa 3061 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 3121 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 3181 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 3241 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 3301 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 3361 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt 3421 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 3481 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 3541 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 3601 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 3661 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 3721 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 3781 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 3841 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 3901 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 3961 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 4021 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 4081 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 4141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 4201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 4261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 4321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 4381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 4441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 4501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 4561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 4621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 4681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 4741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc t //