LOCUS Exported 4196 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION E.coli expression of Cterminal domain of human RanGAP1 with Nterminal His-tag. ACCESSION . VERSION . KEYWORDS pET23a-His-hRanGAP1tail SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4196) AUTHORS Flotho A, Werner A, Winter T, Frank AS, Ehret H, Melchior F TITLE Recombinant reconstitution of sumoylation reactions in vitro. JOURNAL Methods Mol Biol. 2012;832:93-110. doi: 10.1007/978-1-61779-474-2_5. PUBMED 22350878 REFERENCE 2 (bases 1 to 4196) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4196 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 12..467 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(99..118) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 309..330 /label=F1ori-F /note="F1 origin, forward primer" promoter 494..598 /gene="bla" /label=AmpR promoter CDS 599..1459 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(817..836) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1630..2218 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2119..2138 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2372..2389 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2404..2546 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(2472..2491) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2632..2654 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(2648..2839) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind 3348..3367 /label=T7 /note="T7 promoter, forward primer" promoter 3348..3366 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 3398..3420 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 3434..3451 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 4040..4057 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4110..4128) /label=T7 Term /note="T7 terminator, reverse primer" terminator 4124..4171 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" ORIGIN 1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg 1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg 2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat 2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct 2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct 2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt 2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg 2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa 2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc 2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa 3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta 3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg 3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag 3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac 3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca 3301 cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa tacgactcac 3361 tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga 3421 tatacatatg gctcaccacc accaccacca cggtggatcc cctcagcagc gagggcaggg 3481 agagaagtca gccacgccct cacggaagat tctggaccct aacactgggg agccagctcc 3541 cgtgctgtcc tccccacctc ctgcagacgt ctccaccttc ctggcttttc cctctccaga 3601 gaagctgctg cgcctagggc ccaagagctc cgtgctgata gcccagcaga ctgacacgtc 3661 tgaccccgag aaggtggtct ctgccttcct aaaggtgtca tctgtgttca aggacgaagc 3721 tactgtgagg atggcagtgc aggatgcagt agatgccctg atgcagaagg ctttcaactc 3781 ctcgtccttc aactccaaca ccttcctcac caggctgctc gtgcacatgg gtctgctcaa 3841 gagtgaagac aaggtcaagg ccattgccaa cctgtacggc cccctgatgg cgctgaacca 3901 catggtgcag caggactatt tccccaaggc ccttgcaccc ctgctgctgg cgttcgtgac 3961 caagcccaac agcgccctgg aatcctgctc cttcgcccgc cacagtctgc tgcagacgct 4021 gtacaaggtc tagctcgagc accaccacca ccaccactga gatccggctg ctaacaaagc 4081 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg 4141 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggat //