LOCUS Exported 5668 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pBIG2ab - Step2 vector ab (biGBac system for expression of protein complexes in insect cells). ACCESSION . VERSION . KEYWORDS pBIG2ab SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5668) 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 5668) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5668 /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 1364..1818 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1451..1470) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1660..1681 /label=F1ori-F /note="F1 origin, forward primer" promoter 1844..1948 /gene="bla" /label=AmpR promoter CDS 1949..2809 /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(2167..2186) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2980..3568 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3469..3488 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3722..3739 /label=L4440 /note="L4440 vector, forward primer" mobile_element 3873..4097 /mobile_element_type="transposon:Tn7" /note="mini-Tn7 element (right end of the Tn7 transposon)" CDS complement(4164..4697) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" promoter complement(4886..4914) /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" protein_bind 5310..5343 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." mobile_element 5421..5586 /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 tcatgccgtc tgtgatggct tccatgtcgg cagaatgctt 1021 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt 1081 tattggtgcc cttaaacgcc tggtgctacg cctgaataag tgataataag cggatgaatg 1141 gcagaaattc gaaagcaaat tcgacccggt cgtcggttca gggcagggtc gttaaatagc 1201 cgcttatgtc tattgctggt ttaccggttt attgactacc ggaagcagtg tgaccgtgtg 1261 cttctcaaat gcctgaggcc agtttgctca ggctctcccc gtggaggtaa taattgacga 1321 tatgatcatt tattctgcct cccagagcct gatggcgaat gggacgcgcc ctgtagcggc 1381 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc 1441 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc 1501 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc 1561 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg 1621 gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag tggactcttg ttccaaactg 1681 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt 1741 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa 1801 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 1861 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 1921 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat 1981 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt 2041 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag 2101 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa 2161 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg 2221 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct 2281 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac 2341 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca 2401 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat 2461 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact 2521 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc 2581 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga 2641 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg 2701 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg 2761 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca 2821 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta 2881 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca 2941 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 3001 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 3061 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 3121 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 3181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 3241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 3301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct 3361 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc 3421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg 3481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg 3541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct 3601 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga 3661 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 3721 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca 3781 tctgtgcggt atttcacacc gcatagacca gccgcgtaac ctggcaaaat cggttacggt 3841 tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg 3901 aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact 3961 gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa 4021 ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gccaagggca 4081 tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag 4141 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc 4201 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct 4261 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg 4321 agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag 4381 atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc 4441 gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg 4501 taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa 4561 agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg 4621 atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg 4681 ttgctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg 4741 cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aacaagccat 4801 gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg accagttgcg 4861 tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg tcaactgggt 4921 ttgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca gcagcgaagt 4981 cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca cgcatcgtca 5041 ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc tgcccttgct 5101 tcaggagatc ggtagacctc ggccgtcgcg gcgcttgccg gtggtgctga ccccggatga 5161 agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc aggactctag 5221 ctatagttct agtggttggc tacagctttg tttgtactat caacaggttg aactgctgat 5281 caacagatcc tctacgcggc cgcggtacca taacttcgta tagcatacat tatacgaagt 5341 tatctggcct aggttaatta aacgctctat ggtctaaagt ttaaactgga tactattgca 5401 cgtttcaatc aggagatccg aaccagataa gtgaaatcta gttccaaact attttgtcat 5461 ttttaatttt cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt 5521 ccctaaataa tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg 5581 cccacagcgg ggcatttttc ttcctgttat gtttttaatc aaacatcctg ccaactccat 5641 gtgacaaacc gtcatcttcg gctacttt //