LOCUS Exported 6651 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of lamin B receptor. ACCESSION . VERSION . KEYWORDS LBR EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6651) TITLE Lippincott-Schwartz Lab plasmids REFERENCE 2 (bases 1 to 6651) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6651 /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" primer_bind 665..681 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 665..681 /label=pBluescriptSK /note="For pBluescript vector" regulatory 2591..2600 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2597..3316 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2597..3313 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2642..2663) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2903..2922) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3250..3271 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3439..3560 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3476..3495) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3530..3549 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3567..4022) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3704..3725) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3916..3935 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4049..4153 /gene="bla" /label=AmpR promoter promoter 4155..4512 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4178..4198) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4363..4498 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4425..4444 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4547..5341 /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(4601..4620) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5211..5230 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5529..5548) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5573..5620 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5949..6537 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6438..6457 /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 gctagcgcta 601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcggtggcgg 661 ccgctctaga actagtggat cccccgggct gcaggcgccg ggttgctgtg cgactattct 721 ccgggagccg tccgtgtcac cgccggaacc tggcgcaggt taattataga aaatgccaag 781 taggaaattt gccgatggtg aagtggtaag aggtcgatgg cctgggagtt cactttatta 841 tgaagtagaa attctgagcc acgacagcac ctcccagctt tacactgtga agtataaaga 901 tggaacagag cttgaattga aagagaatga tattaagcct ttaacttcct ttaggcaaag 961 gaaaggtggc tcaacttcca gttccccttc cagacgccga gggagtcgat caaggtcacg 1021 ctcccgatcc cctggtcgac cacctaaaag tgcccgccga tctgcttctg cttcccacca 1081 ggccgacatt aaggaagcaa ggagggaagt ggaagttaaa ttgactccgc tgattctgaa 1141 gccatttgga aatagcatca gcagatataa tggggagcct gagcatattg agagaaatga 1201 cgcacctcat aaaaatacac aggaaaaatt cagtttgtca caagaaagca gttacatagc 1261 aacacagtat agccttcgtc caagaagaga agaagtcaaa ttaaaagaaa tagattctaa 1321 ggaagaaaaa tacgttgcaa aagaactggc agtgagaacc tttgaagtga cccccatccg 1381 ggcaaaggac ttggagtttg gaggagtacc tggtgtgttt ctcatcatgt ttggcctgcc 1441 tgtgttcctc ttcctgttgc tgttgatgtg taaacagaaa gatcccagtc ttctgaattt 1501 ccctcctcct ttgccagctt tgtatgagtt atgggaaacc agagtatttg gggtctacct 1561 cctgtggttt ttgattcaag tcctgttcta cctactgcca attggaaagg ttgtagaagg 1621 aacgcctctt attgatggaa gaagactcaa gtatagatta aatggattct atgcttttat 1681 cctgacatct gcagtcatcg gaacatctct cttccagggc gtagagtttc attacgtgta 1741 cagtcatttt cttcagtttg cacttgcggc cactgttttt tgtgtggtct tgagtgtgta 1801 tctctacatg cgctctttga aagcgccccg gaatgacctg tcgcctgcca gctctggaaa 1861 tgctgtctat gatttcttca ttggccgtga attaaaccct cgaattggta cttttgatct 1921 caaatacttt tgtgaattgc gccccggatt gattggatgg gtggttatta acttggtgat 1981 gcttttggct gaaatgaaaa tacaggaccg cgctgttcca tccttggcca tgattttagt 2041 taatagtttc cagcttctct atgtggtgga tgctctctgg aatgaggaag cgttgttgac 2101 gaccatggac atcatccacg atggatttgg attcatgctg gcttttggag acttggtgtg 2161 ggttcccttt atttacagct tccaagcctt ttatttagtc agtcatccaa atgaagtgtc 2221 ttggccaatg gcttctctaa ttattgttct gaaactttgt ggttatgtaa tcttccgagg 2281 tgcaaattct cagaaaaatg cattccggaa aaatcccagt gatccaaagc ttgcacattt 2341 aaaaaccatt catacttcaa cgggaaaaaa tcttctagtt tctggatggt ggggctttgt 2401 tcgccacccc aattacttgg gtgatctcat catggccttg gcgtggtccc tcccatgtgg 2461 ttttaaccac attctgcctt atttctacat aatttatttc accatgttgc ttgtccaccg 2521 agaagctcgt gacgagtacc actgtaagaa gaaatacggc gtggcttggg aaaagtggga 2581 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 2641 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 2701 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 2761 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta 2821 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 2881 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 2941 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 3001 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc 3061 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg 3121 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 3181 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa 3241 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 3301 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta 3361 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 3421 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 3481 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 3541 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc 3601 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 3661 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 3721 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 3781 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 3841 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 3901 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 3961 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 4021 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 4081 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 4141 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 4201 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 4261 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 4321 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 4381 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 4441 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 4501 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 4561 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 4621 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 4681 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 4741 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 4801 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 4861 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 4921 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 4981 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 5041 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 5101 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 5161 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 5221 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 5281 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 5341 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 5401 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 5461 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg 5521 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 5581 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 5641 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 5701 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 5761 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta 5821 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 5881 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 5941 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 6001 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 6061 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 6121 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 6181 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 6241 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 6301 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 6361 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 6421 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 6481 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 6541 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 6601 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t //