LOCUS Exported 7424 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pEGFP-N1 alpha-actinin 1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7424) AUTHORS Edlund M, Lotano MA, Otey CA TITLE Dynamics of alpha-actinin in focal adhesions and stress fibers visualized with alpha-actinin-green fluorescent protein. JOURNAL Cell Motil Cytoskeleton. 2001 Mar . 48(3):190-200. PUBMED 11223950 REFERENCE 2 (bases 1 to 7424) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7424 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 90..393 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 394..597 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 548..568 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 3393..3402 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3399..4118 /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 3399..4115 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3444..3465) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3705..3724) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4052..4073 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 4241..4362 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4278..4297) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4332..4351 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4369..4824) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4506..4527) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4718..4737 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4851..4955 /gene="bla" /label=AmpR promoter promoter 4957..5314 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4980..5000) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5165..5300 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5227..5246 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5349..6143 /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(5403..5422) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6013..6032 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(6331..6350) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6375..6422 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6751..7339 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7240..7259 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca 61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat 481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt 601 ttagtgaacc gtcagatccg ctagcgctac cggactcaga tctcgagctc aagcttcgca 661 ccatcatgga ccattatgat tctcagcaaa ccaacgatta catgcagcca gaagaggact 721 gggaccggga cctgctcctg gacccggcct gggagaagca gcagagaaag acattcacgg 781 catggtgtaa ctcccacctc cggaaggcgg ggacacagat cgagaacatc gaagaggact 841 tccgggatgg cctgaagctc atgctgctgc tggaggtcat ctcaggtgaa cgcttggcca 901 agccagagcg aggcaagatg agagtgcaca agatctccaa cgtcaacaag gccctggatt 961 tcatagccag caaaggcgtc aaactggtgt ccatcggagc cgaagaaatc gtggatggga 1021 atgtgaagat gaccctgggc atgatctgga ccatcatcct gcgctttgcc atccaggaca 1081 tctccgtgga agagacttca gccaaggaag ggctgctcct gtggtgtcag agaaagacag 1141 ccccttacaa aaatgtcaac atccagaact tccacataag ctggaaggat ggcctcggct 1201 tctgtgcttt gatccaccga caccggcccg agctgattga ctacgggaag ctgcggaagg 1261 atgatccact cacaaatctg aatacggctt ttgacgtggc agagaagtac ctggacatcc 1321 ccaagatgct ggatgccgaa gacatcgttg gaactgcccg accggatgag aaagccatca 1381 tgacttacgt gtctagcttc taccacgcct tctctggagc ccagaaggcg gagacagcag 1441 ccaatcgcat ctgcaaggtg ttggccgtca accaggagaa cgagcagctt atggaagact 1501 acgagaagct ggccagtgat ctgttggagt ggatccgccg cacaatcccg tggctggaga 1561 accgggtgcc cgagaacacc atgcatgcca tgcaacagaa gctggaggac ttccgggact 1621 accggcgcct gcacaagccg cccaaggtgc aggagaagtg ccagctggag atcaacttca 1681 acacgctgca gaccaagctg cggctcagca accggcctgc cttcatgccc tctgagggca 1741 ggatggtctc ggacatcaac aatgcctggg gctgcctgga gcaggtggag aagggctatg 1801 aggagtggtt gctgaatgag atccggaggc tggagcgact ggaccacctg gcagagaagt 1861 tccggcagaa ggcctccatc cacgaggcct ggactgacgg caaagaggcc atgctgcgac 1921 agaaggacta tgagaccgcc accctctcgg agatcaaggc cttgctcaag aagcatgagg 1981 ccttcgagag tgacctggct gcccaccagg accgtgtgga gcagattgcc gccatcgcac 2041 aggagctcaa tgagctggac tattatgact cacccagtgt caacgcccgt tgccaaaaga 2101 tctgtgacca gtgggacaat ctgggggccc taactcagaa gcgaagggaa gctctggagc 2161 ggaccgagaa actgctggag accattgacc agctgtactt ggagtatgcc aagcgggctg 2221 cacccttcaa caactggatg gagggggcca tggaggacct gcaggacacc ttcattgtgc 2281 acaccattga ggagatccag ggactgacca cagcccatga gcagttcaag gccaccctcc 2341 ctgatgccga caaggagcgc ctggccatcc tgggcatcca caatgaggtg tccaagattg 2401 tccagaccta ccacgtcaat atggcgggca ccaaccccta cacaaccatc acgcctcagg 2461 agatcaatgg caaatgggac cacgtgcggc agctggtgcc tcggagggac caagctctga 2521 cggaggagca tgcccgacag cagcacaatg agaggctacg caagcagttt ggagcccagg 2581 ccaatgtcat cgggccctgg atccagacca agatggagga gatcgggagg atctccattg 2641 agatgcatgg gaccctggag gaccagctca gccacctgcg gcagtatgag aagagcatcg 2701 tcaactacaa gccaaagatt gatcagctgg agggcgacca ccagctcatc caggaggcgc 2761 tcatcttcga caacaagcac accaactaca ccatggagca catccgtgtg ggctgggagc 2821 agctgctcac caccatcgcc aggaccatca atgaggtaga gaaccagatc ctgacccggg 2881 atgccaaggg catcagccag gagcagatga atgagttccg ggcctccttc aaccactttg 2941 accgggatca ctccggcaca ctgggtcccg aggagttcaa agcctgcctc atcagcttgg 3001 gttatgatat tggcaacgac ccccagggag aagcagaatt tgcccgcatc atgagcattg 3061 tggaccccaa ccgcctgggg gtagtgacat tccaggcctt cattgacttc atgtcccgcg 3121 agacagccga cacagataca gcagaccaag tcatggcttc cttcaagatc ctggctgggg 3181 acaagaacta cattaccatg gacgagctgc gccgcgagct gccacccgac caggctgagt 3241 actgcatcgc gcggatggcc ccctacaccg gccccgactc cgtgccaggt gctctggact 3301 acatgtcctt ctccacggcg ctgtacggcg agagtgacct caagcttcga attctgcagt 3361 cgacggtacc gcgggcccgg gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc 3421 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt 3481 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca 3541 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg 3601 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg 3661 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca 3721 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg 3781 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca 3841 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga 3901 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc 3961 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc 4021 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg 4081 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat 4141 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc 4201 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta 4261 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4321 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg 4381 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat 4441 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg 4501 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc 4561 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt 4621 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag 4681 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg 4741 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc 4801 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc 4861 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 4921 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg 4981 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 5041 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 5101 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 5161 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 5221 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg 5281 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat 5341 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga 5401 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc 5461 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga 5521 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg 5581 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc 5641 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg 5701 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga 5761 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc 5821 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca 5881 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg 5941 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct 6001 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg 6061 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc 6121 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac 6181 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt 6241 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga 6301 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg 6361 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt 6421 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca 6481 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga 6541 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc 6601 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 6661 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 6721 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 6781 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 6841 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 6901 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 6961 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 7021 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 7081 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 7141 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 7201 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 7261 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 7321 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 7381 ctggcctttt gctcacatgt tctttcctgc gttatcccct gatt //