LOCUS Exported 6451 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression of human plus-TIP microtubule binding protein EB3 fused with mEmerald for microtubule +end live tracking. ACCESSION . VERSION . KEYWORDS pCAG mEmerald-EB3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6451) AUTHORS Lewis TL Jr, Turi GF, Kwon SK, Losonczy A, Polleux F TITLE Progressive Decrease of Mitochondrial Motility during Maturation of Cortical Axons In Vitro and In Vivo. JOURNAL Curr Biol. 2016 Oct 10;26(19):2602-2608. doi: 10.1016/j.cub.2016.07.064. Epub 2016 Sep 15. PUBMED 27641765 REFERENCE 2 (bases 1 to 6451) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6451 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(42..629) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(122..141) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(800..1660) /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 1423..1442 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1661..1765) /gene="bla" /label=AmpR promoter enhancer 1796..2175 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2177..2454 /label=chicken beta-actin promoter intron 2455..3467 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 3391..3413 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 3475..3494 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 3555..3564 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 4419..4428 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4425..5144 /codon_start=1 /product="brightly fluorescent monomeric variant of GFP" /label=mEmerald /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(4470..4491) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4731..4750) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 5078..5099 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(5235..5254) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 5300..5355 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(5354..5373) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(5746..5762) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5746..5762) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5759..5781) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5770..5786 /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(5794..5824) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5839..5860 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5919..6114 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5965..6100 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6027..6046 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 6120..6254 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6157..6176) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6211..6230 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6322..6339) /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 61 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 481 agacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 1021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 1081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 1141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 1201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 1261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 1321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 1381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 1441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 1501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 1561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 1621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 1681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 1741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcgacat 1801 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat 1861 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 1921 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 1981 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 2041 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 2101 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 2161 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc 2221 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg 2281 gcgggggggg ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg 2341 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt 2401 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc 2461 gctgcgttgc cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc gccccggctc 2521 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt 2581 aattagcgct tggtttaatg acggctcgtt tcttttctgt ggctgcgtga aagccttaaa 2641 gggctccggg agggcccttt gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt 2701 gtgcgtgggg agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg ctgcgggcgc 2761 ggcgcggggc tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg ggcggtgccc 2821 cgcggtgcgg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg 2881 gtgagcaggg ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac ccccctcccc 2941 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc 3001 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc 3061 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag 3121 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc 3181 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg 3241 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg 3301 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg 3361 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct 3421 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg 3481 tgctggttat tgtgctgtct catcattttg gcaaagaatt gctcgagctc aagcttcgaa 3541 ttctgcagtc gactgccacc atggccgtca atgtgtactc cacatctgtg accagtgaaa 3601 atctgagtcg ccatgatatg cttgcatggg tcaacgactc cctgcacctc aactatacca 3661 agatagaaca gctttgttca ggggcagcct actgccagtt catggacatg ctcttccccg 3721 gctgtgtgca cttgaggaaa gtgaagttcc aggccaaact agagcatgaa tacatccaca 3781 acttcaaggt gctgcaagca gctttcaaga agatgggtgt tgacaaaatc attcctgtag 3841 agaaattagt gaaaggaaaa ttccaagata attttgagtt tattcagtgg tttaagaaat 3901 tctttgacgc aaactatgat ggaaaggatt acaaccctct gctggcgcgg cagggccagg 3961 acgtagcgcc acctcctaac ccaggtgatc agatcttcaa caaatccaag aaactcattg 4021 gcacagcagt tccacagagg acgtccccca caggcccaaa aaacatgcag acctctggcc 4081 ggctgagcaa tgtggccccc ccctgcattc tccggaagaa tcctccatca gcccgaaatg 4141 gcggccatga gactgatgcc caaattcttg aactcaacca acagctggtg gacttgaagc 4201 tgacagtgga tgggctggag aaggaacgtg acttctactt cagcaaactt cgtgacatcg 4261 agctcatctg ccaggagcat gaaagtgaaa acagccctgt tatctcaggc atcattggca 4321 tcctctatgc cacagaggaa ggattcgcac cccctgagga cgatgagatt gaagagcatc 4381 aacaagaaga ccaggacgag tacctggatc caccggtcgc caccatggtg agcaagggcg 4441 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc 4501 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga 4561 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttga 4621 cctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca 4681 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca 4741 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc 4801 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact 4861 acaacagcca caaggtctat atcaccgccg acaagcagaa gaacggcatc aaggtgaact 4921 tcaagacccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga 4981 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt 5041 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga 5101 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagcggcc gcaattcact 5161 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac 5221 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt 5281 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat 5341 tttttgtgtc tatcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat 5401 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt 5461 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat 5521 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct 5581 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga 5641 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcaccgtcga 5701 ccagctggtc gacggtgcac cgtcgaccag cttggcgtaa tcatggtcat agctgtttcc 5761 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 5821 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 5881 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc 5941 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 6001 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct 6061 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta 6121 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 6181 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 6241 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 6301 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 6361 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 6421 cgcaggaaag aacatgtgag caaaaggcca g //