LOCUS sequence-305137- 6159 bp DNA circular SYN 14-APR-2025 DEFINITION Mammalian expression vector for cloning your gene fused to EGFP. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6159) TITLE Fluorescent Protein Plasmids REFERENCE 2 (bases 1 to 6159) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1 https://www.snapgene.com COMMENT SGRef: number: 1; type: "Journal Article" COMMENT Sequence Label: pcDNA3-EGFP FEATURES Location/Qualifiers source 1..6159 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS join(743..745,746..748,749..1462) /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 join(743..745,746..748,749..1459) /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(788..809) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1049..1068) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1396..1417 /label=EGFP-C /note="EGFP, forward primer" promoter complement(1478..1496) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1479..1496) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1516..1533) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1522..1746 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1792..2220 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1879..1898) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2089..2110 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2229..2249) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2234..2563 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2414..2549 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2476..2495 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2630..3424 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2684..2703) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3294..3313 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3598..3719 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3635..3654) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3689..3708 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3768..3784) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3768..3784) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3781..3803) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3792..3808 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(join(3816..3822,3823..3840,3841..3846)) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3861..3882 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3999..4016) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4170..4758) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4250..4269) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(join(4929..5720,5721..5789)) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5552..5571 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5790..5894) /gene="bla" /label=AmpR promoter primer_bind complement(5969..5988) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt 661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattct gcagatatcc 721 atcacactgg cggccgctcg agatggtgag caagggcgag gagctgttca ccggggtggt 781 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 841 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 901 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 961 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 1021 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 1081 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 1141 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 1201 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 1261 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 1321 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 1381 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 1441 catggacgag ctgtacaagt aatctagagg gccctattct atagtgtcac ctaaatgcta 1501 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct 1561 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg 1621 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc 1681 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct 1741 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct 1801 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 1861 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 1921 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 1981 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 2041 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 2101 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 2161 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 2221 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag 2281 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc 2341 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc 2401 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg 2461 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca 2521 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg 2581 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca 2641 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg 2701 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg 2761 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc 2821 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt 2881 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc 2941 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca 3001 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc 3061 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg 3121 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct 3181 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc 3241 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc 3301 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta 3361 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt 3421 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga 3481 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 3541 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 3601 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 3661 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 3721 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 3781 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 3841 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 3901 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 3961 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 4021 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 4081 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 4141 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 4201 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 4261 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 4321 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 4381 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 4441 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 4501 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 4561 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 4621 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 4681 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 4741 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 4801 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 4861 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 4921 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 4981 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 5041 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 5101 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 5161 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 5221 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 5281 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 5341 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 5401 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 5461 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 5521 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 5581 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 5641 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 5701 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 5761 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 5821 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 5881 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 5941 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc 6001 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag 6061 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt 6121 ttgcgctgct tcgcgatgta cgggccagat atacgcgtt //