LOCUS Exported 5596 bp ds-DNA circular SYN 14-4ÔÂ-2018 DEFINITION Baculovirus transfer plasmid with the gp64 promoter, encoding an N-terminal 6xHis-thrombin-S-Tag-enterokinase cassette. ACCESSION . VERSION . KEYWORDS p1064 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5596) AUTHORS Novagen (EMD Millipore) TITLE Direct Submission JOURNAL Exported 2018Äê4ÔÂ14ÈÕ from SnapGene 2.3.2 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5596 /organism="synthetic DNA construct" /lab_host="Insect Cells" /mol_type="other DNA" misc_recomb 329..1156 /note="baculovirus recombination region (lef2/ORF603)" /note="contains ORF603 and part of lef2" CDS 329..453 /codon_start=3 /product="baculovirus late expression factor 2" /note="lef2" /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY" CDS complement(491..1096) /codon_start=1 /product="baculovirus ORF603 protein" /note="ORF603" /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR" promoter 1166..1268 /note="p10 promoter" promoter 1270..1448 /note="gp64 promoter" CDS 1451..1453 /codon_start=1 /product="start codon" /note="ATG" /translation="M" CDS 1457..1474 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 1484..1501 /codon_start=1 /product="thrombin recognition and cleavage site" /note="thrombin site" /translation="LVPRGS" CDS 1505..1549 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /note="S-Tag" /translation="KETAAAKFERQHMDS" CDS 1565..1579 /codon_start=1 /product="enterokinase recognition and cleavage site" /note="enterokinase site" /translation="DDDDK" misc_feature 1581..1645 /note="MCS" /note="multiple cloning site" CDS 1646..1663 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" misc_recomb 1700..3057 /note="baculovirus recombination region (ORF1629)" /note="contains part of ORF1629" CDS complement(1716..3057) /codon_start=2 /product="baculovirus capsid-associated protein" /note="ORF1629" /note="required for viral replication" /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA" rep_origin complement(3320..3908) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4079..4939) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(4940..5044) /gene="bla" /note="AmpR promoter" rep_origin 5071..5526 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" ORIGIN 1 ggaacggctc cgcccactat taatgaaatt aaaaattcca attttaaaaa acgcagcaag 61 agaaacattt gtatgaaaga atgcgtagaa ggaaagaaaa atgtcgtcga catgctgaac 121 aacaagatta atatgcctcc gtgtataaaa aaaatattga acgatttgaa agaaaacaat 181 gtaccgcgcg gcggtatgta caggaagagg tttatactaa actgttacat tgcaaacgtg 241 gtttcgtgtg ccaagtgtga aaaccgatgt ttaatcaagg ctctgacgca tttctacaac 301 cacgactcca agtgtgtggg tgaagtcatg catcttttaa tcaaatccca agatgtgtat 361 aaaccaccaa actgccaaaa aatgaaaact gtcgacaagc tctgtccgtt tgctggcaac 421 tgcaagggtc tcaatcctat ttgtaattat tgaataataa aacaattata aatgctaaat 481 ttgtttttta ttaacgatac aaaccaaacg caacaagaac atttgtagta ttatctataa 541 ttgaaaacgc gtagttataa tcgctgaggt aatatttaaa atcattttca aatgattcac 601 agttaatttg cgacaatata attttatttt cacataaact agacgccttg tcgtcttctt 661 cttcgtattc cttctctttt tcatttttct cttcataaaa attaacatag ttattatcgt 721 atccatatat gtatctatcg tatagagtaa attttttgtt gtcataaata tatatgtctt 781 ttttaatggg gtgtatagta ccgctgcgca tagtttttct gtaatttaca acagtgctat 841 tttctggtag ttcttcggag tgtgttgctt taattattaa atttatataa tcaatgaatt 901 tgggatcgtc ggttttgtac aatatgttgc cggcatagta cgcagcttct tctagttcaa 961 ttacaccatt ttttagcagc accggattaa cataactttc caaaatgttg tacgaaccgt 1021 taaacaaaaa cagttcacct cccttttcta tactattgtc tgcgagcagt tgtttgttgt 1081 taaaaataac agccattgta atgagacgca caaactaata tcacaaactg gaaatgtcta 1141 tcaatatata gttgctgatc agatcccggg acctttaatt caacccaaca caatatatta 1201 tagttaaata agaattatta tcaaatcatt tgtatattaa ttaaaatact atactgtaaa 1261 ttacatttta tcccgttttt ataacagcca gataaaaata atcttatcaa ttaagataaa 1321 aagataagat tattaatcta acaacgtgcc ttgtgtcacg taggccagat aacggtcggg 1381 tatataagac gcctcaacgc tactagtaaa tcagtcacac caaggcttca ataaggaaca 1441 cacaagcacc atggtccatc atcaccacca tcactccgcg ggtttggtgc ccagaggcag 1501 cggcaaagaa acggcggcgg cgaaatttga acgccaacac atggacagcg ctagcggtgg 1561 aggtgatgac gacgacaaga gcccgggctt ctcctcaaaa ggcctggatc cgaattcgag 1621 ctcgaagctt gcggccgcac tcgagcacca ccaccaccac cactaaccta ggtagctgag 1681 cgcatgcaag ctgatccggg ttattagtac atttattaag cgctagattc tgtgcgttgt 1741 tgatttacag acaattgttg tacgtatttt aataattcat taaatttata atctttaggg 1801 tggtatgtta gagcgaaaat caaatgattt tcagcgtctt tatatctgaa tttaaatatt 1861 aaatcctcaa tagatttgta aaataggttt cgattagttt caaacaaggg ttgtttttcc 1921 gaaccgatgg ctggactatc taatggattt tcgctcaacg ccacaaaact tgccaaatct 1981 tgtagcagca atctagcttt gtcgatattc gtttgtgttt tgttttgtaa taaaggttcg 2041 acgtcgttca aaatattatg cgcttttgta tttctttcat cactgtcgtt agtgtacaat 2101 tgactcgacg taaacacgtt aaatagagct tggacatatt taacatcggg cgtgttagct 2161 ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac 2221 gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa 2281 tttgtagttg agctttttgg aattatttct gattgcgggc gtttttgggc gggtttcaat 2341 ctaactgtgc ccgattttaa ttcagacaac acgttagaaa gcgatggtgc aggcggtggt 2401 aacatttcag acggcaaatc tactaatggc ggcggtggtg gagctgatga taaatctacc 2461 atcggtggag gcgcaggcgg ggctggcggc ggaggcggag gcggaggtgg tggcggtgat 2521 gcagacggcg gtttaggctc aaatgtctct ttaggcaaca cagtcggcac ctcaactatt 2581 gtactggttt cgggcgccgt ttttggtttg accggtctga gacgagtgcg atttttttcg 2641 tttctaatag cttccaacaa ttgttgtctg tcgtctaaag gtgcagcggg ttgaggttcc 2701 gtcggcattg gtggagcggg cggcaattca gacatcgatg gtggtggtgg tggtggaggc 2761 gctggaatgt taggcacggg agaaggtggt ggcggcggtg ccgccggtat aatttgttct 2821 ggtttagttt gttcgcgcac gattgtgggc accggcgcag gcgccgctgg ctgcacaacg 2881 gaaggtcgtc tgcttcgagg cagcgcttgg ggtggtggca attcaatatt ataattggaa 2941 tacaaatcgt aaaaatctgc tataagcatt gtaatttcgc tatcgtttac cgtgccgata 3001 tttaacaacc gctcaatgta agcaattgta ttgtaaagag attgtctcaa gctcggatcg 3061 atcccgcacg ccgataacaa gccttttcat ttttactaca gcattgtagt ggcgagacac 3121 ttcgctgtcg tcgaggttta aacgcttcct cgctcactga ctcgctgcgc tcggtcgttc 3181 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 3241 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 3301 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 3361 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 3421 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 3481 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 3541 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 3601 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 3661 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 3721 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg 3781 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 3841 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 3901 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 3961 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 4021 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 4081 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 4141 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 4201 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 4261 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 4321 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 4381 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 4441 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 4501 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 4561 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 4621 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 4681 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 4741 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 4801 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 4861 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 4921 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 4981 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 5041 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg 5101 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 5161 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 5221 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 5281 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 5341 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 5401 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 5461 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta 5521 caatttccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct 5581 cttcgctatt acgcca //