LOCUS pCAGGS-Flag-N 4804 bp ds-DNA circular SYN 23-AUG-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4804) AUTHOR Name | HonorGene: http://www.honorgene.com AUTHOR Tel | 15574926881 | AUTHOR Email | 12695920@qq.com | FEATURES Location/Qualifiers source 1..4804 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 385..662 /label=chicken beta-actin promoter intron 663..1680 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" CDS 1738..1761 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" misc_feature 1762..1811 /label=MCS polyA_site 1891..1946 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" protein_bind 2331..2347 /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(2355..2385) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2400..2421 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2480..2675 /label=SV40 promoter /note="SV40 early promoter" rep_origin 2526..2661 /label=SV40 ori /note="SV40 origin of replication" polyA_signal 2681..2815 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(3053..3641) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3812..4672) /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" promoter complement(4673..4777) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca 421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag 481 cgatgggggc gggggggggg ggggggcccc ccccaggcgg ggcggggcgg ggcgaggggc 541 ggggcggggc gaggcggaaa ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt 601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg 661 cgggagtcgt tgcgcgctgc cttccccccg tgccccgctc cgccgccgcc tcgcgccgcc 721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc 781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg 841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt 901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc 961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg 1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg 1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc 1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc 1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg 1261 cggggccgcc tcgggccggg gagggctcgg gggaaggggc gcggcggccc ccggagcgcc 1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg 1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc 1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg 1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt 1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt 1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag 1681 ctcctgggca acgtgctggt tgttgtgctg tctcatcatt ttggcaaagc caccatggat 1741 tacaaggatg acgacgataa ggaattcgag ctcatcgatg gtacccgggc atgcctcgag 1801 ctagcagatc ttaattaatt aaggatcttt ttccctctgc caaaaattat ggggacatca 1861 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag 1921 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa 1981 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca 2041 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt 2101 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt 2161 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc 2221 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc 2281 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 2341 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 2401 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 2461 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc 2521 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 2581 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 2641 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta 2701 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 2761 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg 2821 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 2881 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 2941 tcaaaggcgg taatacggtt atccacagaa tcagggataa cgcaggaaag aacatgtgag 3001 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 3061 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 3121 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 3181 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 3241 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 3301 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 3361 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 3421 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 3481 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 3541 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 3601 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 3661 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 3721 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 3781 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 3841 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 3901 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 3961 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 4021 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 4081 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 4141 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 4201 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 4261 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 4321 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 4381 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 4441 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 4501 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 4561 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 4621 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 4681 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 4741 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 4801 ctgg //