LOCUS Exported 6800 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3.1-P25C-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6800) TITLE Li-Huei Tsai's Plasmids. REFERENCE 2 (bases 1 to 6800) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6800 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..30) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 202..581 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 582..785 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 736..756 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 830..849 /label=T7 /note="T7 promoter, forward primer" promoter 830..848 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1591..2301 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELY" primer_bind complement(1633..1654) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1894..1913) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2241..2262 /label=EGFP-C /note="EGFP, forward primer" promoter complement(2320..2338) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2321..2338) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2358..2375) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2364..2588 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2634..3062 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2721..2740) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2931..2952 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3071..3091) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3076..3405 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3256..3391 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3318..3337 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3472..4266 /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(3526..3545) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4136..4155 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4440..4561 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4477..4496) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4531..4550 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4610..4626) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4610..4626) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4623..4645) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4634..4650 /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(4658..4688) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4703..4724 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4841..4858) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5012..5600) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5092..5111) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5771..6631) /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 6394..6413 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6632..6736) /gene="bla" /label=AmpR promoter ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt 901 gctggaattc tgcagatggg atccaccatg gcccagcccc caccggccca gccgcctgca 961 cccccggcca gccagctctc gggttcccag accgggggct cctcctcagt caagaaagcc 1021 cctcaccctg ccgtcacctc cgcagggacg cccaaacggg tcatcgtcca ggcgtccacc 1081 agtgagctgc ttcgctgcct gggtgagttt ctctgccgcc ggtgctaccg cctgaagcac 1141 ctgtccccca cggaccccgt gctctggctg cgcagcgtgg accgctcgct gcttctgcag 1201 ggctggcagg accagggctt catcacgccg gccaacgtgg tcttcctcta catgctctgc 1261 agggatgtta tctcctccga ggtgggctcg gatcacgagc tccaggccgt cctgctgaca 1321 tgcctgtacc tctcctactc ctacatgggc aacgagatct cctacccgct caagcccttc 1381 ctggtggaga gctgcaagga ggccttttgg gaccgttgcc tctctgtcat caacctcatg 1441 agctcaaaga tgctgcagat aaatgccgac ccacactact tcacacaggt cttctccgac 1501 ctgaagaacg agagcggcca ggaggacaag aagcggctcc tcctaggcct ggatcggtta 1561 ggggatccga gctcggtacc aagcttggtg gtgagcaagg gcgaggagct gttcaccggg 1621 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc 1681 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc 1741 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc 1801 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa 1861 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc 1921 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc 1981 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc 2041 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac 2101 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac 2161 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac 2221 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact 2281 ctcggcatgg acgagctgta ctagtctaga gggccctatt ctatagtgtc acctaaatgc 2341 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 2401 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 2461 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 2521 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 2581 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 2641 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 2701 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 2761 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 2821 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2881 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2941 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 3001 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 3061 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 3121 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 3181 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 3241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 3301 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 3361 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 3421 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 3481 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 3541 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 3601 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 3661 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 3721 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 3781 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3841 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3901 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3961 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 4021 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 4081 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 4141 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 4201 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 4261 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 4321 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 4381 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 4441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 4501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 4561 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 4621 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 4681 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 4741 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 4801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 5041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 5101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 5161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 5221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 5281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 5341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 5401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 5581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 5821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 5881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 5941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 6001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 6061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 6121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 6181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 6241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 6301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 6361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6721 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag 6781 atctcccgat cccctatggt //