LOCUS Exported 5669 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pBIG2abc - Step2 vector abc (biGBac system for expression of protein complexes in insect cells). ACCESSION . VERSION . KEYWORDS pBIG2abc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5669) 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 5669) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5669 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 303..405 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 406..1065 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(472..491) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" rep_origin 1365..1819 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1452..1471) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1661..1682 /label=F1ori-F /note="F1 origin, forward primer" promoter 1845..1949 /gene="bla" /label=AmpR promoter CDS 1950..2810 /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(2168..2187) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2981..3569 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3470..3489 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3723..3740 /label=L4440 /note="L4440 vector, forward primer" mobile_element 3874..4098 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (right end of the Tn7 transposon)" CDS complement(4165..4698) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(4887..4915) /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" protein_bind 5311..5344 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." mobile_element 5422..5587 /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 ctgaacagga gggacagctg atagaaacag 121 aagccactgg agcacctcaa aaacaccatc atacactaaa tcagtaagtt ggcagcatca 181 cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc agaataaata 241 aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg aaaatgagac 301 gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc 361 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc 421 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt 481 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta 541 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc 601 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg 661 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc 721 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg 781 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc 841 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac 901 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg 961 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt 1021 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt 1081 tattggtgcc cttaaacgcc tggttgctac gcctgaataa gtgataataa gcggatgaat 1141 ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt cgttaaatag 1201 ccgcttatgt ctattgctgg tttaccggtt tattgactac cggaagcagt gtgaccgtgt 1261 gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta ataattgacg 1321 atatgatcat ttattctgcc tcccagagcc tgatggcgaa tgggacgcgc cctgtagcgg 1381 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 1441 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 1501 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 1561 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 1621 ggtttttcgc cctttgacgt tggagtccac gttcttaata gtggactctt gttccaaact 1681 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt 1741 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa 1801 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 1861 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 1921 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta 1981 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag 2041 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca 2101 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta 2161 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc 2221 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc 2281 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca 2341 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc 2401 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca 2461 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac 2521 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg 2581 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg 2641 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg 2701 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac 2761 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc 2821 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 2881 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 2941 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 3001 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 3061 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 3121 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 3181 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 3241 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 3301 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 3361 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 3421 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 3481 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 3541 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 3601 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 3661 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 3721 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc 3781 atctgtgcgg tatttcacac cgcatagacc agccgcgtaa cctggcaaaa tcggttacgg 3841 ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa gtcttaaact 3901 gaacaaaata gatctaaact atgacaataa agtcttaaac tagacagaat agttgtaaac 3961 tgaaatcagt ccagttatgc tgtgaaaaag catactggac ttttgttatg gctaaagcaa 4021 actcttcatt ttctgaagtg caaattgccc gtcgtattaa agaggggcgt ggccaagggc 4081 atggtaaaga ctatattcgc ggcgttgtga caatttaccg aacaactccg cggccgggaa 4141 gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat 4201 cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc 4261 tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat 4321 gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat catagatata 4381 gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag cgccaacaac 4441 cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca agttcccgag 4501 gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact cacgaccgaa 4561 aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac atgtgcgaat 4621 gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct gcgtaacatc 4681 gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac 4741 gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca 4801 tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc 4861 gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg 4921 tttgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag 4981 tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc 5041 aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgcccttgc 5101 ttcaggagat cggtagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg 5161 aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc caggactcta 5221 gctatagttc tagtggttgg ctacagcttt gtttgtacta tcaacaggtt gaactgctga 5281 tcaacagatc ctctacgcgg ccgcggtacc ataacttcgt atagcataca ttatacgaag 5341 ttatctggcc taggttaatt aaacgctcta tggtctaaag tttaaaccta atgatgcctg 5401 atgtttcaat caggagatcc gaaccagata agtgaaatct agttccaaac tattttgtca 5461 tttttaattt tcgtattagc ttacgacgct acacccagtt cccatctatt ttgtcactct 5521 tccctaaata atccttaaaa actccatttc cacccctccc agttcccaac tattttgtcc 5581 gcccacagcg gggcattttt cttcctgtta tgtttttaat caaacatcct gccaactcca 5641 tgtgacaaac cgtcatcttc ggctacttt //