LOCUS Exported 8335 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Venus-PSD-95 in mammalian cells. ACCESSION . VERSION . KEYWORDS pCMV-5U-Venus-PSD-95-3U SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8335) AUTHORS Ifrim MF, Williams KR, Bassell GJ TITLE Single-Molecule Imaging of PSD-95 mRNA Translation in Dendrites and Its Dysregulation in a Mouse Model of Fragile X Syndrome. JOURNAL J Neurosci. 2015 May 6;35(18):7116-30. doi: 10.1523/JNEUROSCI.2802-14.2015. PUBMED 25948262 REFERENCE 2 (bases 1 to 8335) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8335 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 57..360 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 361..564 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 515..535 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 914..923 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 950..1666 /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(995..1016) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1256..1275) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1603..1624 /label=EGFP-C /note="EGFP, forward primer" regulatory 1673..1682 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 5119..5240 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5156..5175) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5210..5229 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(5247..5702) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5384..5405) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5596..5615 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5729..5833 /gene="bla" /label=AmpR promoter promoter 5835..6192 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5858..5878) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 6043..6178 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6105..6124 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6227..7021 /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(6281..6300) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6891..6910 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(7209..7228) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 7253..7300 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 7629..8217 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8118..8137 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt 61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg 361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt 421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg 541 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gccagctcat 601 gccccagccc cagaactccg gggctgccag ccccctgtgc ccctacccct cctgagaatc 661 ggggtgtgct cctcaagccc cctcccctcc actggggagc cccttttagg gtcccccctc 721 ccttccacgc tcccctaccc ttctcttctc caaccttctc cttcccctct cacccctccc 781 cccatcctgg ccccccctgc aaaagtgggg tgcggagggg ggaggggtga gaacccaccg 841 aggggaggaa caaaactcca atgaagtcag agccccctac tcgccgccgc ggccaggccc 901 cccaacaccg gtcgccacca tggactgtct ctgtatagtg acaaccaaga tggtgagcaa 961 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa 1021 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac 1081 cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac 1141 cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt 1201 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga 1261 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat 1321 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta 1381 caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc 1441 caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg accactacca 1501 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta 1561 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt 1621 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg cggccgccat 1681 ggactgtctc tgtatagtga caaccaagaa ataccgctac caagatgaag acacgccccc 1741 tctggaacac agcccggccc acctccccaa ccaggccaat tctccccctg tgattgtcaa 1801 cacggacacc ctagaagccc caggatatgt gaacggaaca gagggggaga tggagtatga 1861 ggagatcaca ttggaaaggg gtaactcagg tctgggcttc agcatcgcag gtggcaccga 1921 caacccacac atcggtgacg acccatccat ctttatcacc aagatcattc ctggtggggc 1981 tgcagcccag gatggccgcc tcagggtcaa cgacagcatc ctgtttgtca atgaagtgga 2041 tgtccgggag gtgacccatt cagctgcagt ggaggccctc aaagaggcgg gttccatcgt 2101 tcgcctctac gtcatgcgcc ggaaaccccc agctgagaag atcatagaga tcaagcttat 2161 caaagggcct aaaggacttg gcttcagcat cgcagggggc gttgggaacc agcacatccc 2221 tggagataat agcatctacg taaccaagat catcgaagga ggcgctgccc acaaggatgg 2281 caggttgcag atcggagaca agatcctggc ggtcaacagt gtggggctag aggatgtcat 2341 gcatgaggac gccgtggcag ccctgaagaa cacatatgac gttgtgtacc taaaggtggc 2401 caagcccagc aatgcctacc tgagtgacag ctatgctccc ccagacatca caacctcata 2461 ttctcagcac ctggacaatg agatcagtca tagcagctac ttgggcaccg actaccccac 2521 agccatgacc cccacttccc ctcggcgcta ctcccccgtg gccaaggacc tgctagggga 2581 ggaagatatt ccccgggaac caaggcggat cgtgatccat cggggctcca ccggcctggg 2641 cttcaacatt gtgggcggcg aggacggtga aggcatcttc atctccttca tccttgctgg 2701 gggtccagct gacctcagtg gggagctacg gaagggggac cagatcctgt cggtcaatgg 2761 tgttgacctc cgcaatgcca gtcatgaaca ggctgccatt gccctgaaga acgcgggtca 2821 gacggtcacg atcatcgctc agtataaacc agaagagtat agccgattcg aggccaagat 2881 ccatgatctt cgggaacagc ttatgaatag tagcctgggc tcagggactg catctctgcg 2941 aagcaacccc aagcggggct tctatatcag ggccctgttt gactacgaca agaccaagga 3001 ctgcggtttc ttgagccagg ccctgagctt ccactttggg gatgtgcttc atgtaattga 3061 cgccagcgac gaagagtggt ggcaagcgcg gcgggtccac tctgacagtg agaccgatga 3121 cattggcttc attcccagca aacggcgggt cgagcgacga gagtggtcaa ggttaaaggc 3181 caaggactgg ggctccagct ctggatcaca gggtcgagaa gactcggttc tgagctatga 3241 gacggtgacg cagatggaag tgcactacgc tcgccccatc atcatccttg ggcctaccaa 3301 agaccgtgcc aacgatgatc ttctctccga gttccccgac aagtttggat cctgtgtccc 3361 tcatacgaca cgtcctaagc gggaatatga gatagacggc cgcgattacc actttgtctc 3421 ctcccgggag aaaatggaga aggacattca ggcgcacaag ttcattgagg ctggccagta 3481 caacagccac ctctacggga ccagcgtcca gtctgtgcga gaggtagcag agcaggggaa 3541 gcactgcatc cttgatgtct cagccaatgc cgtgcggcgg ctgcaggcgg cccacctgca 3601 ccctatcgcc atcttcatcc gtccccgctc cctggagaat gtgctagaga tcaataagcg 3661 gatcacagag gagcaagccc ggaaagcctt cgacagggcc acgaagctgg agcaggagtt 3721 cacggagtgc ttctcagcca tcgtagaggg cgacagcttt gaagagatct atcacaaagt 3781 gaaacgtgtc atcgaagacc tctcaggccc ctacatctgg gtcccagccc gagagagact 3841 ctgaactagt gggatccgcc ctgaaaaagg gctcgagccc tgaaaaaggg caattgccct 3901 gaaaaagggc gtcgacgccc tgaaaaaggg cagatctaac gttttcctgc cctggctggg 3961 cctggactca ccctgcctcc accacctggg cccttggtct ggactgaata gcccaagccc 4021 ttcgtaccct agcctttctt cccacccttc ttatttattt cctttctgac tggatcgagc 4081 ccattggagg gaggacactc ctctctgcat gtatccctgc accccagatc agggccccta 4141 gaccccagga acctggggtc tgggggggag ctgggctcct ggttccgagc ccttgctcct 4201 taagactcct acccctgccc gcccccatgc acacacagac ccactggggg cctctgccct 4261 ccccattttc tcccacacac attccagaag tcagggcccc tctcaaggag tacccgctgt 4321 agggatgcag ggccacaggc ttccgcgctc tcccgaggca ggggctgggg tcacccctgc 4381 cccatcataa ctccccacgc cgtttgagtt ctcctttatt ttctccattc ttttcttctc 4441 aaaggtggtt tttgggggga gaagcagggg ggctctcctg agggtccccc cgtctttcac 4501 acacctccca ccttttttct ttgccggttt gcatgagtgg aaggtctaac tgtggctttt 4561 ttttttttct gggattttat ttcattttat ttgattttat ttttttggga aaaggggagg 4621 gatgggtctg gggagtgggg aatgtgggag gggggtgggg gggcaaaggg tcaggggttg 4681 ggtgtccggg agccagggga ggacaggaaa tgctgccgcc ttctgcaatt tatttatttt 4741 tttcttttga gagagtgaaa ggaagagaca gacacttgaa taaaggtacc agcttgtggc 4801 tctgtggctt tgctgggtgt ggggagcagg ttgggtgggt gtggagctct attggggggg 4861 gcatctaggg tgggctaggc cttgctgatg gtatctagtg ggagggctac aggcccttta 4921 atgaagtctc acaaataacc acaccttcag actgtcttga agagttgaaa aggtactcct 4981 ccatccggaa gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta 5041 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 5101 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 5161 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 5221 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc 5281 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 5341 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 5401 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 5461 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 5521 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 5581 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 5641 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 5701 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 5761 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 5821 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 5881 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 5941 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 6001 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 6061 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6121 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6181 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 6241 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 6301 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 6361 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 6421 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 6481 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 6541 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 6601 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 6661 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 6721 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 6781 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 6841 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 6901 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 6961 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 7021 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 7081 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 7141 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg 7201 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 7261 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 7321 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 7381 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 7441 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta 7501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 7561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 7621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 7681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 7741 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 7801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 7861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 7921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 7981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 8041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 8101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 8161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 8221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 8281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagt //