LOCUS Exported 3300 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR221_EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3300) AUTHORS Yang X, Boehm JS, Yang X, Salehi-Ashtiani K, Hao T, Shen Y, Lubonja R, Thomas SR, Alkan O, Bhimdi T, Green TM, Johannessen CM, Silver SJ, Nguyen C, Murray RR, Hieronymus H, Balcha D, Fan C, Lin C, Ghamsari L, Vidal M, Hahn WC, Hill DE, Root DE TITLE A public genome-scale lentiviral expression library of human ORFs. JOURNAL Nat Methods. 2011 Jun 26. ():. PUBMED 21706014 REFERENCE 2 (bases 1 to 3300) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3300 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1..717 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(46..67) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(307..326) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 654..675 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(737..836) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(853..872) /label=T7 /note="T7 promoter, forward primer" promoter complement(854..872) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(877..893) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(877..893) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(995..1014) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1006..1815 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1073..1092) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 1962..2550 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2451..2470 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2704..2721 /label=L4440 /note="L4440 vector, forward primer" terminator 2880..2907 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 2999..3085 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 3134..3156 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3148..3165 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3149..3165 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 3181..3280 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" ORIGIN 1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac 61 ggcgacgtaa acggccacaa gttcagcgtg tctggcgagg gcgagggcga tgccacctac 121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc 181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag 241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc 301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg 361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac 421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac 481 ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc 541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac 601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc 661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc 721 aagggtgggc gcgccgaccc agctttcttg tacaaagttg gcattataag aaagcattgc 781 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat 841 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc 901 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga 961 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa 1021 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa 1081 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc 1141 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat 1201 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt 1261 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc 1321 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc 1381 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt 1441 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat 1501 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca 1561 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac 1621 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag 1681 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt 1741 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc 1801 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg 1861 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc 1921 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 1981 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 2041 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 2101 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 2161 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 2221 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 2281 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 2341 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 2401 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 2461 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 2521 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 2581 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 2641 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 2701 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc 2761 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc 2821 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa 2881 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc 2941 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat 3001 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt 3061 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta 3121 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc 3181 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa 3241 tgctttttta taatgccaac tttgtacaaa aaagcaggct ccgcggccgc ccccttcacc //