LOCUS Exported 5067 bp ds-DNA circular SYN 14-FEB-2023 DEFINITION Mammalian expression with secretion signal sequence. ACCESSION . VERSION . KEYWORDS pHL-sec SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5067) AUTHORS Aricescu AR, Lu W, Jones EY TITLE A time- and cost-efficient system for high-level protein production in mammalian cells. JOURNAL Acta Crystallogr D Biol Crystallogr. 2006 Oct;62(Pt 10):1243-50. Epub 2006 Sep 19. PUBMED 17001101 REFERENCE 2 (bases 1 to 5067) AUTHORS . TITLE Direct Submission JOURNAL Exported Feb 14, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5067 /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..1679 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1603..1625 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1687..1706 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 1761..1770 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2292..2309 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2421..2440) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2486..2541 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2540..2559) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(2902..2918) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2902..2918) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2915..2937) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2926..2942 /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(2950..2980) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2995..3016 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3145..3162) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3316..3904) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3396..3415) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4075..4935) /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 4698..4717 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4936..5040) /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 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc 541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt 601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg 661 cgggagtcgc tgcgcgctgc cttcgccccg 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 gggaggggcg cggcggcccc cggagcgccg 1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg 1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca 1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg 1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc 1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg 1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc 1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttgcggccgt 1741 ctcaggccga attcaagctt gccaccatgg ggatccttcc cagccctggg atgcctgcgc 1801 tgctctccct cgtgagcctt ctctccgtgc tgctgatggg ttgcgtagct gaaaccggtt 1861 cgatagtttt agagcctatc tattggaatt cctcgaactc caaatttcta cctggacaag 1921 gactggtact atacccacag ataggagaca aattggatat tatttgcccc aaagtggact 1981 ctaaaactgt tggccagtat gaatattata aagtttatat ggttgataaa gaccaagcag 2041 acagatgcac tattaagaag gaaaataccc ctctcctcaa ctgtgccaaa ccggaccaag 2101 atatcaaatt caccatcaag tttcaagaat tcagccctaa cctctggggt ctagaatttc 2161 agaagaacaa agattattac attatatcta catcaaatgg gtctttggag ggcctggata 2221 accaggaggg aggggtgtgc cagacaagag ccatgaagat cctcatgaaa gttggacaag 2281 atggtaccaa gcaccaccat caccatcact aatgatcact cgagactagt atcgcgataa 2341 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg 2401 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag 2461 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt 2521 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat 2581 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac 2641 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta 2701 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt 2761 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc 2821 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag 2881 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 2941 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 3001 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 3061 gtgccagcgg atcgatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 3121 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct 3181 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga 3241 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc 3301 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg 3361 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg 3421 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt 3481 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt 3541 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg 3601 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact 3661 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt 3721 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct 3781 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac 3841 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc 3901 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg 3961 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 4021 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 4081 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 4141 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 4201 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 4261 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 4321 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 4381 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 4441 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 4501 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 4561 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 4621 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 4681 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 4741 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 4801 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 4861 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 4921 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 4981 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg 5041 cacatttccc cgaaaagtgc cacctgg //