LOCUS Exported 5915 bp ds-DNA circular SYN 12-JUL-2021 DEFINITION pBIG1a - Step1 vector a (biGBac system for expression of protein complexes in insect cells). ACCESSION . VERSION . KEYWORDS pBIG1a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5915) AUTHORS Weissmann F, Petzold G, VanderLinden R, Huis In 't Veld PJ, Brown NG, Lampert F, Westermann S, Stark H, Schulman BA, Peters JM TITLE biGBac enables rapid gene assembly for the expression of large multisubunit protein complexes. JOURNAL Proc Natl Acad Sci U S A. 2016 May 10;113(19):E2564-9. doi: 10.1073/pnas.1604935113. Epub 2016 Apr 25. PUBMED 27114506 REFERENCE 2 (bases 1 to 5915) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5915 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 442..1233 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 1274..1728 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1361..1380) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1570..1591 /label=F1ori-F /note="F1 origin, forward primer" promoter 1754..1858 /gene="bla" /label=AmpR promoter CDS 1859..2719 /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 complement(2077..2096) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2890..3478 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3379..3398 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3632..3649 /label=L4440 /note="L4440 vector, forward primer" mobile_element 3783..4007 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (right end of the Tn7 transposon)" CDS complement(4074..4607) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(4796..4824) /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" protein_bind 5220..5253 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(5299..5318) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5343..5391 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" mobile_element 5668..5833 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (left end of the Tn7 transposon)" ORIGIN 1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga 61 ctgaatatca acgcttattt gcagcctgaa ctgggaggca gaataaatga tcatatcgtc 121 aattattacc tccacgggga gagcctgagc aaactggcct caggcatttg agaagcacac 181 ggtcacactg cttccggtag tcaataaacc ggtaagtagc gtatgcgctc acgcaactgg 241 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt 301 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac 361 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc 421 cctaaaacaa agttaaacat catgagggaa gcggtgatcg ccgaagtatc gactcaacta 481 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg 541 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg 601 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact 661 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac 721 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag 781 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc 841 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc 901 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg 961 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt 1021 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg 1081 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga 1141 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg 1201 aaaggcgaga tcaccaaggt agtcggcaaa taatgtctaa caattcgttc aagccgacgg 1261 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 1321 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 1381 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 1441 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 1501 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 1561 ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt 1621 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 1681 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt 1741 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 1801 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 1861 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 1921 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 1981 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 2041 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 2101 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 2161 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 2221 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 2281 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 2341 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 2401 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 2461 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 2521 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 2581 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 2641 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 2701 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 2761 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 2821 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 2881 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 2941 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 3001 aactggcttc agcagagcgc agataccaaa tactattctt ctagtgtagc cgtagttagg 3061 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 3121 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 3181 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 3241 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 3301 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 3361 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 3421 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 3481 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 3541 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 3601 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 3661 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca 3721 gccgcgtaac ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg 3781 ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa 3841 gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc 3901 atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg 3961 tcgtattaaa gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac 4021 aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg 4081 gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag 4141 agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc 4201 gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc 4261 tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc 4321 agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg 4381 aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag 4441 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact 4501 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt 4561 ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca 4621 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga 4681 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct 4741 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt 4801 acgaaccgaa caggcttatg tcaactgggt ttgtgccttc atccgtttcc acggtgtgcg 4861 tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga 4921 gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt tcttctacgg 4981 caaggtgctg tgcacggatc tgcccttgct tcaggagatc ggtagacctc ggccgtcgcg 5041 gcgcttgccg gtggtgctga ccccggatga agtggttcgc atcctcggtt ttctggaagg 5101 cgagcatcgt ttgttcgccc aggactctag ctatagttct agtggttggc tacagctttg 5161 tttgtactat caacaggttg aactgctgat caacagatcc tctacgcggc cgcggtacca 5221 taacttcgta tagcatacat tatacgaagt tatctgggtt taaacgctct atggtctaaa 5281 gtttttatcg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat 5341 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg 5401 gggttcggtc ccagggctgg cactctgtca agtgaacgct ctatggtcta aagatttaaa 5461 tctatatctc aatcggggtt acgtcgatac cccaccgaga ccccattggg accaatacgc 5521 ccgcgtttct tccttttccc caccccaacc cccaagttcg ggtgaaggcc cagggctcgc 5581 agccaacgtc ggggcggcaa gccctgccat agccactacg ggttaattaa aaactatatc 5641 tcaatcgggg tttaaacagg agatccgaac cagataagtg aaatctagtt ccaaactatt 5701 ttgtcatttt taattttcgt attagcttac gacgctacac ccagttccca tctattttgt 5761 cactcttccc taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt 5821 ttgtccgccc acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca 5881 actccatgtg acaaaccgtc atcttcggct acttt //