LOCUS Exported 4738 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses N-terminal cysteine Abeta(1-40). ACCESSION . VERSION . KEYWORDS pET-Sac-Abeta(MC1-40) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4738) TITLE An efficient expression system for N-terminal cysteine A-beta for bioconjugation JOURNAL Unpublished REFERENCE 2 (bases 1 to 4738) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4738 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..20 /label=T7 /note="T7 promoter, forward primer" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 51..73 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" primer_bind complement(268..286) /label=T7 Term /note="T7 terminator, reverse primer" terminator 282..329 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter complement(695..723) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" primer_bind complement(750..768) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 836..940 /gene="bla" /label=AmpR promoter CDS 941..1801 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1159..1178) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1972..2560 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2461..2480 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2714..2731 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2746..2885 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2811..2830) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2971..2993 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2987..3178) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 4636..4655 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac 61 tttaagaagg agatatacat atgtgcgacg ctgaattccg tcacgactct ggttacgaag 121 ttcaccacca gaagctggtg ttcttcgctg aagacgtggg ttctaacaag ggtgctatca 181 tcggtctgat ggttggtggc gttgtgtaat aggagctcga tccggctgct aacaaagccc 241 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg 301 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac 361 aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc 421 aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat 481 tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg 541 acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc 601 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga 661 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt 721 caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa 781 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg 841 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 901 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 961 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 1021 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 1081 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 1141 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga 1201 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 1261 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 1321 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 1381 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 1441 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac 1501 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 1561 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 1621 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 1681 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 1741 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 1801 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 1861 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 1921 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 1981 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 2041 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 2101 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 2161 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 2221 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 2281 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 2341 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 2401 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 2461 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 2521 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 2581 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 2641 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 2701 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt 2761 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc 2821 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg 2881 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 2941 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 3001 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg 3061 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta 3121 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg 3181 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga 3241 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg 3301 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc 3361 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc 3421 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac 3481 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc 3541 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc 3601 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc 3661 tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc 3721 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag 3781 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca 3841 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc 3901 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat 3961 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc 4021 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag 4081 aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc 4141 cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc 4201 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag 4261 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc 4321 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt 4381 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg 4441 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt 4501 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc 4561 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc 4621 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc 4681 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaat //