LOCUS Exported 5411 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS YFP-NavII-III SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5411) AUTHORS Grubb MS, Burrone J TITLE Activity-dependent relocation of the axon initial segment fine-tunes neuronal excitability. JOURNAL Nature. 2010 Jun 24. 465(7301):1070-4. PUBMED 20543823 REFERENCE 2 (bases 1 to 5411) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5411 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 490..509 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 764..1067 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1068..1271 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1222..1242 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 1294..1374 /label=MCS /note="multiple cloning site" regulatory 1376..1385 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1382..2098 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1427..1448) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1688..1707) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2035..2056 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2902..3023 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2939..2958) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2993..3012 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3030..3485) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3167..3188) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3379..3398 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3512..3616 /gene="bla" /label=AmpR promoter promoter 3618..3975 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3641..3661) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3826..3961 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3888..3907 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4010..4804 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4064..4083) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4674..4693 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4992..5011) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5036..5083 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" ORIGIN 1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat 721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg 781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt 841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac 901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg 961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact 1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt 1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc 1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc 1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata 1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga 1321 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac 1381 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga 1441 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta 1501 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac 1561 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa 1621 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt 1681 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct 1741 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca 1801 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa 1861 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc 1921 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca 1981 ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt 2041 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagag 2101 ttctttcagc tcagacaacc tggctgccac agatgacgat aacgaaatga acaacctcca 2161 gatagccgtg ggaaggatgc agaagggaat tgattttgtt aaaaggaaga tacgtgaatt 2221 cattcagaaa gcctttgtca gaaagcagaa agctttagat gaaatcaaac cgctggaaga 2281 tctgaataac aagaaagaca gttgtatctc caaccacacg accatagaaa taggcaagga 2341 cctcaattac ctcaaagatg gaaacgggac gaccagtggc ataggcagca gtgtggagaa 2401 gtatgtggta gatgagagtg attacatgtc gttcataaac aaccccagcc tcaccgtgac 2461 tgtgcccatc gcccttggag agtctgactt tgaaaactta aacacggaag aattcagtag 2521 tgagtcagat atggaagaaa gcaaggagaa attgaatgca actagctcat ctgagggcag 2581 tacggttgat ataggcgctc cggcagaggg agagcagcca gaggccgaac cggaagaatc 2641 gcttgaaccg gaagcgtgtt tcacagaaga ctgtgtgaga aagttcaagt gttgtcagat 2701 aagcatagaa gagggcaaag gcaagctctg gtggaacttg aggaaaacgt gctacaagat 2761 aagctagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt 2821 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 2881 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 2941 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 3001 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt 3061 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 3121 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 3181 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 3241 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 3301 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 3361 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 3421 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 3481 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta 3541 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata 3601 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg 3661 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3721 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 3781 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact 3841 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag 3901 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc 3961 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca 4021 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 4081 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 4141 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc 4201 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 4261 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 4321 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 4381 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 4441 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 4501 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct 4561 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 4621 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 4681 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 4741 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 4801 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 4861 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 4921 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg 4981 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca 5041 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg 5101 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc 5161 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc 5221 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat 5281 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 5341 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 5401 aaaggatctt c //