LOCUS Exported 6944 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 6944) 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 6944) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6944 /organism="synthetic DNA construct" /mol_type="other DNA" intron 228..1244 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1168..1190 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1252..1271 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1332..1341 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 2196..2205 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2202..2921 /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(2247..2268) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2508..2527) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2855..2876 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3012..3031) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3077..3132 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3131..3150) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(3523..3539) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3523..3539) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3536..3558) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3547..3563 /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(3571..3601) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3616..3637 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3696..3891 /label=SV40 promoter /note="SV40 early promoter" rep_origin 3742..3877 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3804..3823 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 3897..4031 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3934..3953) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3988..4007 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4099..4116) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4270..4858) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4350..4369) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5029..5889) /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 5652..5671 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5890..5994) /gene="bla" /label=AmpR promoter enhancer 6025..6404 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 ccccccnatt tttttttttt tttttttttt tattttttgg gggggggggg gggggggggg 61 gggggggggg gcgcgcgccg ggsggggcgg ggcggggcga ggggcggggc ggggcgaggc 121 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga 181 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc 241 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac 301 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt 361 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc 421 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg 481 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg 541 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc 601 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt 661 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag 721 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg 781 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg 841 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg 901 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt 961 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc 1021 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt 1081 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct 1141 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta 1201 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc 1261 tggttattgt gctgtctcat cattttggca aagaattgct cgagctcaag cttcgaattc 1321 tgcagtcgac tgccaccatg gccgtcaatg tgtactccac atctgtgacc agtgaaaatc 1381 tgagtcgcca tgatatgctt gcatgggtca acgactccct gcacctcaac tataccaaga 1441 tagaacagct ttgttcaggg gcagcctact gccagttcat ggacatgctc ttccccggct 1501 gtgtgcactt gaggaaagtg aagttccagg ccaaactaga gcatgaatac atccacaact 1561 tcaaggtgct gcaagcagct ttcaagaaga tgggtgttga caaaatcatt cctgtagaga 1621 aattagtgaa aggaaaattc caagataatt ttgagtttat tcagtggttt aagaaattct 1681 ttgacgcaaa ctatgatgga aaggattaca accctctgct ggcgcggcag ggccaggacg 1741 tagcgccacc tcctaaccca ggtgatcaga tcttcaacaa atccaagaaa ctcattggca 1801 cagcagttcc acagaggacg tcccccacag gcccaaaaaa catgcagacc tctggccggc 1861 tgagcaatgt ggcccccccc tgcattctcc ggaagaatcc tccatcagcc cgaaatggcg 1921 gccatgagac tgatgcccaa attcttgaac tcaaccaaca gctggtggac ttgaagctga 1981 cagtggatgg gctggagaag gaacgtgact tctacttcag caaacttcgt gacatcgagc 2041 tcatctgcca ggagcatgaa agtgaaaaca gccctgttat ctcaggcatc attggcatcc 2101 tctatgccac agaggaagga ttcgcacccc ctgaggacga tgagattgaa gagcatcaac 2161 aagaagacca ggacgagtac ctggatccac cggtcgccac catggtgagc aagggcgagg 2221 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca 2281 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt 2341 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttgacct 2401 acggcgtgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt 2461 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact 2521 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga 2581 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca 2641 acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag gtgaacttca 2701 agacccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca 2761 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtcca 2821 agctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg 2881 ccgccgggat cactctcggc atggacgagc tgtacaagta agcggccgca attcactcct 2941 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa 3001 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag 3061 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt 3121 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag 3181 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg 3241 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga 3301 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta 3361 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta 3421 ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca ccgtcgacca 3481 gctggtcgac ggtgcaccgt cgaccagctt ggcgtaatca tggtcatagc tgtttcctgt 3541 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 3601 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 3661 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata 3721 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 3781 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag 3841 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact 3901 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 3961 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 4021 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 4081 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 4141 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 4201 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 4261 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 4321 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 4381 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 4441 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 4501 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 4561 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 4621 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 4681 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 4741 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 4801 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 4861 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 4921 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 4981 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 5041 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 5101 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 5161 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 5221 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 5281 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 5341 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 5401 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 5461 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 5521 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 5581 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 5641 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 5701 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 5761 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 5821 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 5881 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 5941 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6001 tccccgaaaa gtgccacctg ggtcgacatt gattattgac tagttattaa tagtaatcaa 6061 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa 6121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 6181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 6241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 6301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 6361 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca 6421 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta 6481 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgg gcgccmgggg 6541 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg 6601 nggggggggg ggggnggggg ggggggnggg gggggggggg gggggggggg gggggggggg 6661 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg 6721 gggggggggg gggggggggg gggggggggg nggggggggg gggggggggg gggggggggg 6781 gggbgggggg gggggggggg gggggggggg agcagggggt gtgggcgcgt cggtcgggct 6841 gcaacccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg 6901 gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc gggg //