LOCUS Exported 4888 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pSKBAD2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4888) AUTHORS Iwai H, Zuger S, Jin J, Tam PH TITLE Highly efficient protein trans-splicing by a naturally split DnaE intein from Nostoc punctiforme. JOURNAL FEBS Lett. 2006 Mar 20. 580(7):1853-8. PUBMED 16516207 REFERENCE 2 (bases 1 to 4888) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4888 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(329..346) /label=pBAD Reverse /note="For vectors with E. coli araBAD promoter, reverse primer" terminator 498..584 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 676..703 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" promoter 721..812 /gene="bla" /label=AmpR promoter CDS 813..1673 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1031..1050) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1715..2170 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1802..1821) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2012..2033 /label=F1ori-F /note="F1 origin, forward primer" rep_origin 2281..2869 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2770..2789 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3023..3040 /label=L4440 /note="L4440 vector, forward primer" misc_feature 3055..3195 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3121..3140) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3281..3303 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3442..3461 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" CDS complement(3644..4522) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /label=araC /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 4549..4833 /gene="araBAD" /label=araBAD promoter /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction (Guzman et al., 1995)" primer_bind 4756..4775 /label=pBAD Forward /note="For vectors with E. coli araBAD promoter, forward primer" RBS 4860..4882 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" ORIGIN 1 tatgatcaaa atagccacac gtaaatattt aggcaaacaa aatgtctatg acattggagt 61 tgagcgcgac cataattttg cactcaaaaa tggcttcata gcttctaatt gtttcaatgg 121 tacccagtac aaacttatcc tgaacggtaa aaccctgaaa ggtgaaacca ccaccgaagc 181 tgttgacgct gctaccgcgg aaaaagtttt caaacagtac gctaacgaca acggtgttga 241 cggtgaatgg acctacgacg acgctaccaa aacctacacc gtaacggaat aagcttggct 301 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg 361 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc 421 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtccccc atgcgagagt 481 agggaactgc caggcatcaa ataaaacgaa aggctcagtg caaagactgg gcctttcgtt 541 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt 601 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca 661 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc 721 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 781 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc 841 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga 901 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca 961 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt 1021 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg 1081 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc 1141 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata 1201 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt 1261 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag 1321 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca 1381 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg 1441 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg 1501 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag 1561 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg 1621 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag 1681 accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg cgcattaagc 1741 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 1801 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 1861 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 1921 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 1981 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 2041 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 2101 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg 2161 tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 2221 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 2281 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 2341 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 2401 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 2461 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 2521 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 2581 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 2641 ctacaccgaa ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg 2701 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 2761 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 2821 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 2881 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc 2941 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg 3001 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat 3061 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cggatatggt gcactctcag 3121 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac 3181 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 3241 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 3301 aggttttcac cgtcatcacc gaaacgcgcg aggcagcaag gagatggcgc ccaacagtcc 3361 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg 3421 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt 3481 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atctaattct catgtttgac 3541 agcttatcat cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc 3601 tatgctactc cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg 3661 ctacatcatt cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc 3721 attttttaaa tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg 3781 tggcgatagg catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct 3841 cgcgccagct taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg 3901 gcgacaagca aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat 3961 cgctgatgta ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt 4021 taatcgcttc catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg 4081 aatagcgccc ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg 4141 gctggtgcgc ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa 4201 gccattcatg ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag 4261 cctccggatg acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg 4321 gtcggcaaac aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat 4381 tgagaatata acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg 4441 cctcaatcgg cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg 4501 gatcattttg cgcttcagcc atacttttca tactcccgcc attcagagaa gaaaccaatt 4561 gtccatattg catcagacat tgccgtcact gcgtctttta ctggctcttc tcgctaacca 4621 aaccggtaac cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa 4681 aaacgcgtaa caaaagtgtc tataatcacg gcagaaaagt ccacattgat tatttgcacg 4741 gcgtcacact ttgctatgcc atagcatttt tatccataag attagcggat cctacctgac 4801 gctttttatc gcaactctct actgtttctc catacccgtt tttttgggct agaaataatt 4861 ttgtttaact ttaagaagga gatataca //