LOCUS Exported 5614 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS CMV::SypHy A4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5614) AUTHORS Granseth B, Odermatt B, Royle SJ, Lagnado L TITLE Clathrin-mediated endocytosis is the dominant mechanism of vesicle retrieval at hippocampal synapses. JOURNAL Neuron. 2006 Sep 21. 51(6):773-86. PUBMED 16982422 REFERENCE 2 (bases 1 to 5614) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5614 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 602..611 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1166..1876 /codon_start=1 /product="pH-sensitive mutant of green fluorescent protein (Ng et al., 2002)" /label=superecliptic pHluorin /note="excited strongly by blue light at neutral pH but minimally fluorescent at acidic pH" /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" primer_bind complement(1193..1221) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 1816..1836 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" polyA_signal 2402..2523 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2439..2458) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2493..2512 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2530..2985) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2667..2688) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2879..2898 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3012..3116 /gene="bla" /label=AmpR promoter promoter 3118..3475 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3141..3161) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3326..3461 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3388..3407 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3510..4304 /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(3564..3583) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4174..4193 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4492..4511) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4536..4583 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4912..5500 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5401..5420 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcctcg 601 agccaccatg gacgtggtga atcagctggt ggctgggggt cagttccggg tggtcaagga 661 gccccttggc ttcgtgaagg tgctgcagtg ggtctttgcc atcttcgcct ttgctacgtg 721 tggcagctac accggggagc ttcggctgag cgtggagtgt gccaacaaga cggagagtgc 781 cctcaacatc gaagttgaat tcgagtaccc cttcaggctg caccaagtgt actttgatgc 841 accctcctgc gtcaaagggg gcactaccaa gatcttcctg gttggggact actcctcgtc 901 ggctgaattc tttgtcaccg tggctgtgtt tgccttcctc tactccatgg gggccctggc 961 cacctacatc ttcctgcaga acaagtaccg agagaacaac aaagggccta tgatggactt 1021 tctggctaca gccgtgttcg ctttcatgtg gctagttagt tcatcagcct gggccaaagg 1081 cctgtccgat gtgaagatgg ccacggaccc agagaacatt atcaaggaga tgcccatgtg 1141 ccgccagaca gggaacaccg gtggaagtaa aggagaagaa cttttcactg gagttgtccc 1201 aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg 1261 tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact 1321 acctgttcct tggccaacac ttgtcactac tttaacttat ggtgttcaat gcttttcaag 1381 atacccagat catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt 1441 acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg ctgaagtcaa 1501 gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga 1561 tggaaacatt cttggacaca aattggaata caactataac gatcaccagg tgtacatcat 1621 ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attagacaca acattgaaga 1681 tggaggcgtt caactagcag accattatca acaaaatact ccaattggcg atgggcccgt 1741 ccttttacca gacaaccatt acctgtttac aacttctact ctttcgaaag atcccaacga 1801 aaagagagac cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat 1861 ggatgaacta tacaaaaccg gtcgccagac agggaacaca tgcaaggaac tgagggaccc 1921 tgtgacttca ggactcaaca ccacagtggt gtttggcttc ctgaacctgg tgctctgggt 1981 tggcaactta tggttcgtgt tcaaggagac aggctgggca gccccattca tgcgcgcacc 2041 tccaggcgcc ccggaaaagc aaccagcacc tggcgatgcc tacggcgatg cgggctacgg 2101 gcagggcccc ggaggctatg ggccccagga ctcctacggg cctcagggtg gttatcaacc 2161 cgattacggg cagccagcca gcggtggcgg tggctacggg cctcagggcg actatgggca 2221 gcaaggctat ggccaacagg gtgcgcccac ctccttctcc aatcagatgt aatctgcaga 2281 ttaatctaga taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt 2341 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 2401 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 2461 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 2521 ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa 2581 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat 2641 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg 2701 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac 2761 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta 2821 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag 2881 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg 2941 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc 3001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 3061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg 3121 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc 3181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa 3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 3301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc 3361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc 3421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg 3481 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 3541 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 3601 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 3661 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg 3721 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3781 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 3841 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 3901 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 3961 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 4021 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 4081 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 4141 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4201 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4261 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4321 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 4381 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 4441 gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg 4501 gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac 4561 gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt 4621 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc 4681 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc 4741 tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt 4801 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 4861 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 4921 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 4981 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 5041 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 5101 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 5161 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 5221 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 5281 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 5341 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 5401 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 5461 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 5521 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 5581 tgattctgtg gataaccgta ttaccgccat gcat //