LOCUS Exported 6159 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression vector for cloning your gene fused to CFP. ACCESSION . VERSION . KEYWORDS pcDNA3-CFP 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6159 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(32..51) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 223..602 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 603..806 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 757..777 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 851..870 /label=T7 /note="T7 promoter, forward primer" promoter 851..869 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 965..1684 /codon_start=1 /product="cyan variant of GFP" /label=CFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1010..1031) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1271..1290) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1618..1639 /label=EGFP-C /note="EGFP, forward primer" promoter complement(1700..1718) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1701..1718) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1738..1755) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1744..1968 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2014..2442 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2101..2120) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2311..2332 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2451..2471) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2456..2785 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2636..2771 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2698..2717 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2852..3646 /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(2906..2925) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3516..3535 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3820..3941 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3857..3876) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3911..3930 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3990..4006) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3990..4006) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4003..4025) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4014..4030 /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(4038..4068) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4083..4104 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4221..4238) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4392..4980) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4472..4491) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5151..6011) /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 5774..5793 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6012..6116) /gene="bla" /label=AmpR promoter ORIGIN 1 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa 61 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt 121 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc 181 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta 241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 601 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat 661 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 721 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac 781 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc 841 ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc 901 actagtaacg gccgccagtg tgctggaatt ctgcagatat ccatcacact ggcggccgct 961 cgagatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct 1021 ggacggcgac gtaaacggcc acaggttcag cgtgtccggc gagggcgagg gcgatgccac 1081 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc 1141 caccctcgtg accaccctga cctggggcgt gcagtgcttc agccgctacc ccgaccacat 1201 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgtaccat 1261 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac 1321 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg 1381 gcacaagctg gagtacaact acatcagcca caacgtctat atcaccgccg acaagcagaa 1441 gaacggcatc aaggcccact tcaagatccg ccacaacatc gaggacggca gcgtgcagct 1501 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa 1561 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat 1621 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa 1681 gtaatctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct 1741 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga 1801 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt 1861 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg 1921 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg 1981 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg 2041 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 2101 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 2161 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 2221 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 2281 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac 2341 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt 2401 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg 2461 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 2521 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat 2581 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc 2641 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat 2701 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt 2761 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc 2821 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 2881 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 2941 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 3001 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct 3061 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 3121 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 3181 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 3241 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 3301 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 3361 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 3421 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 3481 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 3541 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 3601 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 3661 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 3721 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 3781 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat 3841 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 3901 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg 3961 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat 4021 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc 4081 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga 4141 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt 4201 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg 4261 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac 4321 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg 4381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca 4441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc 4501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc 4561 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag 4621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc 4681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca 4741 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg 4801 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg 4861 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct 4921 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 4981 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 5041 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 5101 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 5161 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 5221 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 5281 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 5341 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 5401 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 5461 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 5521 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 5581 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 5641 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 5701 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 5761 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 5821 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 5881 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 5941 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 6001 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 6061 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 6121 tttccccgaa aagtgccacc tgacgtcgac ggatcggga //