LOCUS Exported 7088 bp ds-DNA circular SYN 18-APR-2017 DEFINITION . ACCESSION . VERSION . KEYWORDS pEF1a-BirA-V5-neo SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7088) AUTHORS . TITLE Direct Submission JOURNAL Exported Tuesday, Jul 25, 2017 from SnapGene Viewer 2.5.0 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7088 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 18..59 /codon_start=1 /product="epitope tag from simian virus 5" /note="V5 tag" /translation="GKPIPNPLLGLDST" CDS 69..86 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" polyA_signal 115..339 /note="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin 385..813 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 827..1156 /note="SV40 promoter" /note="SV40 enhancer and early promoter" rep_origin 1007..1142 /note="SV40 ori" /note="SV40 origin of replication" CDS 1223..2017 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="NeoR/KanR" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 2193..2314 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(2363..2379) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind 2387..2403 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2411..2441) /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 2456..2477 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(2754..3342) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3513..4373) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(4374..4478) /gene="bla" /note="AmpR promoter" promoter 4860..6038 /note="EF-1-alpha promoter" /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 5091..6029 /note="EF-1-alpha intron A" /note="intron upstream of the start codon of human EF-1-alpha" promoter 6055..6073 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 6125..7087 /codon_start=1 /gene="birA" /product="E. coli biotin protein ligase" /note="BirA" /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK" ORIGIN 1 ctagagggcc cttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc 61 gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc 121 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag 181 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta 241 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag 301 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca 361 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg 421 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg 481 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 541 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 601 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 661 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta 721 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa 781 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg 841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 901 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 961 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 1021 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 1081 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 1141 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga 1201 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 1261 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 1321 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 1381 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac 1441 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 1501 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 1561 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 1621 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 1681 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 1741 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 1801 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 1861 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 1921 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 1981 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa 2041 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 2101 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 2161 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 2221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 2281 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 2341 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 2401 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 2461 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 2521 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 2581 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 2641 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 2701 agaacatgtg agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 2761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 2821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 2881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 2941 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 3001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 3061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 3121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 3181 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 3241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 3301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 3361 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 3421 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 3481 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 3541 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 3601 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 3661 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 3721 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 3781 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 3841 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 3901 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 3961 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 4021 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 4081 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 4141 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 4201 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 4261 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 4321 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 4381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 4441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 4501 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 4561 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 4621 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 4681 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 4741 gttgacattg attattgact aggcttttgc aaaaagcttt gcaaagatgg ataaagtttt 4801 aaacagagag gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg 4861 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg 4921 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg 4981 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag 5041 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg 5101 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta 5161 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg 5221 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc 5281 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt 5341 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc 5401 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg 5461 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag 5521 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg 5581 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag 5641 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga 5701 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt 5761 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt 5821 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg 5881 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat 5941 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag 6001 tggttcaaag tttttttctt ccatttcagg tgtcgtgagg aattagcttg gtactaatac 6061 gactcactat agggagaccc aagctggcta ggtaagcttg gtaccgagct cggatccgcc 6121 agccatgaag gataacaccg tgccactgaa attgattgcc ctgttagcga acggtgaatt 6181 tcactctggc gagcagttgg gtgaaacgct gggaatgagc cgggcggcta ttaataaaca 6241 cattcagaca ctgcgtgact ggggcgttga tgtctttacc gttccgggta aaggatacag 6301 cctgcctgag cctatccagt tacttaatgc taaacagata ttgggtcagc tggatggcgg 6361 tagtgtagcc gtgctgccag tgattgactc cacgaatcag taccttcttg atcgtatcgg 6421 agagcttaaa tcgggcgatg cttgcattgc agaataccag caggctggcc gtggtcgccg 6481 gggtcggaaa tggttttcgc cttttggcgc aaacttatat ttgtcgatgt tctggcgtct 6541 ggaacaaggc ccggcggcgg cgattggttt aagtctggtt atcggtatcg tgatggcgga 6601 agtattacgc aagctgggtg cagataaagt tcgtgttaaa tggcctaatg acctctatct 6661 gcaggatcgc aagctggcag gcattctggt ggagctgact ggcaaaactg gcgatgcggc 6721 gcaaatagtc attggagccg ggatcaacat ggcaatgcgc cgtgttgaag agagtgtcgt 6781 taatcagggg tggatcacgc tgcaggaagc ggggatcaat ctcgatcgta atacgttggc 6841 ggccatgcta atacgtgaat tacgtgctgc gttggaactc ttcgaacaag aaggattggc 6901 accttatctg tcgcgctggg aaaagctgga taattttatt aatcgcccag tgaaacttat 6961 cattggtgat aaagaaatat ttggcatttc acgcggaata gacaaacagg gggctttatt 7021 acttgagcag gatggaataa taaaaccctg gatgggcggt gaaatatccc tgcgtagtgc 7081 agaaaaat //