LOCUS Exported 5801 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of GFP-FKBP at the plasma membrane in mammalian cells. Plasma membrane targeting is via CD8 alpha chain.. ACCESSION . VERSION . KEYWORDS CD8-GFP-FKBP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5801) AUTHORS Clarke NI, Royle SJ TITLE FerriTag is a new genetically-encoded inducible tag for correlative light-electron microscopy. JOURNAL Nat Commun. 2018 Jul 4;9(1):2604. doi: 10.1038/s41467-018-04993-0. PUBMED 29973588 REFERENCE 2 (bases 1 to 5801) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5801 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 59..362 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 363..566 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 517..537 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 1380..2090 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELY" CDS 1380..2090 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELY" primer_bind complement(1422..1443) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1683..1702) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2030..2051 /label=EGFP-C /note="EGFP, forward primer" CDS 2139..2465 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL LKLE" polyA_signal 2587..2708 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2624..2643) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2678..2697 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2715..3170) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2852..2873) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3064..3083 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3197..3301 /gene="bla" /label=AmpR promoter promoter 3303..3660 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3326..3346) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3511..3646 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3573..3592 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3695..4489 /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(3749..3768) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4359..4378 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4677..4696) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4721..4768 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5097..5685 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5586..5605 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc 601 ggactcagat ctcgagctca agcttcgaat tcggcggctc ccgcgcgcct cccctcgcgc 661 ccgagcttcg agccaagcag cgtcctgggg agcgcgtcat ggccttacca gtgaccgcct 721 tgctcctgcc gctggccttg ctgctccacg ccgccaggcc gagccagttc cgggtgtcgc 781 cgctggatcg gacctggaac ctgggcgaga cagtggagct gaagtgccag gtgctgctgt 841 ccaacccgac gtcgggctgc tcgtggctct tccagccgcg cggcgccgcc gccagtccca 901 ccttcctcct atacctctcc caaaacaagc ccaaggcggc cgaggggctg gacacccagc 961 ggttctcggg caagaggttg ggggacacct tcgtcctcac cctgagcgac ttccgccgag 1021 agaacgaggg ctactatttc tgctcggccc tgagcaactc catcatgtac ttcagccact 1081 tcgtgccggt cttcctgcca gcgaagccca ccacgacgcc agcgccgcga ccaccaacac 1141 cggcgcccac catcgcgtcg cagcccctgt ccctgcgccc agaggcgtgc cggccagcgg 1201 cggggggcgc agtgcacacg agggggctgg acttcgcctg tgatatctac atctgggcgc 1261 ccttggccgg gacttgtgga gtccttctcc tgtcactggt tatcaccctt tactgcaagc 1321 ggcttaagcg aagaaggatc cccgccgccg ccgccgccgc cgccgccgtc agaccggtgg 1381 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg 1441 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca 1501 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg 1561 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc 1621 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca 1681 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga 1741 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc 1801 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca 1861 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc 1921 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc 1981 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc 2041 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac cgaatcgatg 2101 tgaatggggg ccctgaaccc gccaagaacc tgtacaccat gggagtgcag gtggaaacca 2161 tctccccagg agacgggcgc accttcccca agcgcggcca gacctgcgtg gtgcactaca 2221 ccgggatgct tgaagatgga aagaaatttg attcctcccg ggacagaaac aagcccttta 2281 agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt gcccagatga 2341 gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt gccactgggc 2401 acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt ctaaaactgg 2461 aatgagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt 2521 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 2581 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 2641 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 2701 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg 2761 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa 2821 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa 2881 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg 2941 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa 3001 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa 3061 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct 3121 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac 3181 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat 3241 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag 3301 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca 3361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt 3421 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 3481 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc 3541 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg 3601 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa 3661 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 3721 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 3781 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 3841 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg 3901 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 3961 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 4021 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 4081 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 4141 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 4201 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg 4261 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 4321 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 4381 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 4441 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 4501 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 4561 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 4621 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa 4681 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat 4741 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac 4801 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc 4861 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca 4921 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt 4981 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa 5041 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 5101 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 5161 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 5221 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag 5281 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 5341 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 5401 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 5461 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 5521 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 5581 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 5641 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 5701 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 5761 tcccctgatt ctgtggataa ccgtattacc gccatgcatt a //