LOCUS pET-15b-HopQAD-FLAG 6896 bp DNA circular SYN 06-SEP-2025 DEFINITION . ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 6896) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 6, 2025 from SnapGene 8.1.1 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6896 /mol_type="other DNA" /organism="recombinant plasmid" promoter 104..208 /gene="bla" /label=AmpR promoter /note="color: #ffffff" CDS 209..1069 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="color: #993366" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1240..1828 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" misc_feature 2014..2156 /label=bom /note="basis of mobility region from pBR322" /note="color: #a6acb3" CDS complement(2258..2449) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /note="color: #993366" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" protein_bind complement(3727..3748) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." /note="color: #31849b; direction: LEFT" CDS complement(3761..4843) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /note="color: #993366" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(4844..4921) /gene="lacI (mutant)" /label=lacIq promoter /note="In the lacIq allele, a single base change in the promoter boosts expression of the lacI gene about 10-fold." /note="color: #ffffff; direction: LEFT" promoter 5230..5248 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff" protein_bind 5249..5273 /label=lac operator /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)." /note="color: #31849b" primer_bind complement(5273..5316) /label=tagVec-R /note="color: 黑色; sequence: ggtatatctccttcttaaagttaaacaaaattatttctagaggg; added: 2025-08-28" RBS 5288..5310 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" /note="color: #a6acb3" primer_bind 5297..5345 /label=tagHQ-F /note="color: 黑色; sequence: ctttaagaaggagatataccatggctgtacaaaaagttaaaaatgcaga; added: 2025-08-28" CDS join(5317..6594,6595..6597) /label=HopQAD /note="This forward directional feature has 2 segments: 1: 5317 .. 6594 / #00ff00 2: 6595 .. 6597 / #00ff00" primer_bind 5317..5345 /label=tagHop-F /note="color: 黑色; sequence: CCGCACTCGAatggctgtacaaaaagttaaaaatgcaga; added: 2025-08-27" misc_feature join(5320..6594,6595..6597) /label=Insert Sequence /note="Insert Sequence" /note="This feature has 2 segments: 1: 5320 .. 6594 / #a6acb3 2: 6595 .. 6597 / #a6acb3" primer_bind complement(6574..6594) /label=tagHopQ-R /note="color: 黑色; sequence: attgttggtggtcgaactggc; added: 2025-08-28" CDS 6598..6621 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /note="color: #cc99b2" /translation="DYKDDDDK" primer_bind 6602..6649 /label=tagVec-F /note="color: 黑色; sequence: ACAAAGACGACGACGATAAAtaagctgagcaataactagcataacccc; added: 2025-08-28" misc_feature 6622..6624 /label=Insert Sequence /note="Insert Sequence" /note="color: #a6acb3" terminator 6637..6684 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" /note="color: #ffffff" promoter complement(6859..6887) /label=tet promoter /note="E. coli promoter for tetracycline efflux protein gene" /note="color: #ffffff; direction: LEFT" ORIGIN 1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat 61 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 121 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 181 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 241 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 301 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 361 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 421 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg 481 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 541 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 601 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 661 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 721 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact 781 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 841 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 901 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 961 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 1021 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 1081 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 1141 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 1201 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 1261 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 1321 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 1381 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 1441 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 1501 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 1561 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 1621 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 1681 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 1741 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 1801 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 1861 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 1921 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 1981 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 2041 tctgtgcggt atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc 2101 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc 2161 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt 2221 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac 2281 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga 2341 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc 2401 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg 2461 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca 2521 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac 2581 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg 2641 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga 2701 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga 2761 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc 2821 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg 2881 tcctcaacga caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga 2941 tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg 3001 tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc 3061 cgttagcgag gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg 3121 acgcaacgcg gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt 3181 ccatgtgctc gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt 3241 taggctggta agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg 3301 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat 3361 aatggggaag gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc 3421 ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt 3481 gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat 3541 cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg 3601 tcctacgagt tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg 3661 cgcccaccgg aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg 3721 tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc 3781 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt 3841 gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc 3901 ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca 3961 ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat 4021 cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc 4081 gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct 4141 cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt 4201 ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac 4261 gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga 4321 ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg 4381 gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag 4441 caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga 4501 gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca 4561 ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg 4621 cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt 4681 gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc 4741 gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga 4801 caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt 4861 gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt 4921 ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg 4981 ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac 5041 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg 5101 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca 5161 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc 5221 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag 5281 aaataatttt gtttaacttt aagaaggaga tataccatgg ctgtacaaaa agttaaaaat 5341 gcagataagg tgcagaagct gtccgacacg tacgagcagt tgagtcgcct gttaaccaac 5401 gataacggaa ccaacagcaa aacttccgcg caggcgatca accaagcagt gaacaacctg 5461 aacgaacgtg cgaaaaccct ggccggcggt acgacgaact ctccggcata ccaggctacc 5521 ttgctcgcgc tgcgtagcgt gctgggtctt tggaatagca tgggttatgc cgtcatctgc 5581 ggtggctaca ccaaatcccc gggtgagaat aatcagaagg acttccacta tactgatgaa 5641 aacggcaatg gcactacgat taactgcggc ggtagcacca actcaaatgg cacccacagc 5701 tataatggta ccaacaccct gaaggctgac aaaaatgtta gcctgtccat tgaacaatat 5761 gagaaaatcc acgaggcata ccagatccta tcgaaagcgc tgaagcaggc aggcctggct 5821 ccgctgaaca gcaagggtga gaagctggaa gcgcatgtta ccacgagcaa gtatcaacaa 5881 gataaccaga ccaagaccac caccagcgtt attgatacca cgaacgacgc tcaaaacctg 5941 ctgacccaag cgcaaacgat cgtaaatacc ctgaaggact actgcccgat tctgattgca 6001 aagagctctt cctccaatgg cggcacaaac aacgcgaaca ccccaagctg gcagactgcg 6061 ggtggtggta aaaactcttg tgctaccttt ggtgcggaat tcagcgccgc gagcgacatg 6121 atcaacaacg cgcaaaaaat cgtgcaagag acccagcaac tgtccgccaa tcagccgaaa 6181 aatattaccc aaccgcataa tcttaacttg aattccccga gctctctgac tgccttggcc 6241 caaaaaatgc tgaagaacgc gcagagccag gcggagatcc tgaagttggc gaaccaggtc 6301 gagtccgact ttaacaaatt gtctagcggt catttgaagg attacattgg taaatgtgat 6361 gcttccgcga ttagctcggc aaacatgacc atgcagaacc agaaaaacaa ctggggtaac 6421 gggtgcgcgg gtgttgaaga aacccaatcc ctcctgaaaa cgagcgcggc agacttcaat 6481 aatcaaaccc cgcagattaa tcaagcccag aatctcgcta acacgttgat ccaggagctg 6541 ggcaataacc cgtttcgtaa tatgggcatg atcgccagtt cgaccaccaa caatggcgac 6601 tacaaagacg acgacgataa ataagctgag caataactag cataacccct tggggcctct 6661 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata tcccgcaaga 6721 ggcccggcag taccggcata accaagccta tgcctacagc atccagggtg acggtgccga 6781 ggatgacgat gagcgcattg ttagatttca tacacggtgc ctgactgcgt tagcaattta 6841 actgtgataa actaccgcat taaagcttat cgatgataag ctgtcaaaca tgagaa 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