LOCUS Exported 6831 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS FLAG-p54 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6831) AUTHORS Rosonina E, Ip JY, Calarco JA, Bakowski MA, Emili A, McCracken S, Tucker P, Ingles CJ, Blencowe BJ TITLE Role for PSF in mediating transcriptional activator-dependent stimulation of pre-mRNA processing in vivo. JOURNAL Mol Cell Biol. 2005 Aug;25(15):6734-46. PUBMED 16024807 REFERENCE 2 (bases 1 to 6831) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6831 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(63..80) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(234..822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(314..333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(993..1853) /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 1616..1635 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1854..1958) /gene="bla" /label=AmpR promoter primer_bind complement(2033..2052) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 2224..2603 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2604..2807 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2758..2778 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2852..2871 /label=T7 /note="T7 promoter, forward primer" promoter 2852..2870 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 2885..2894 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2894..2917 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(4373..4391) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4374..4391) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(4411..4428) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4417..4641 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4687..5115 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4774..4793) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4984..5005 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5124..5144) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5129..5458 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5309..5444 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5371..5390 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5525..6319 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5579..5598) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6189..6208 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6493..6614 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6530..6549) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6584..6603 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6663..6679) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6663..6679) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6676..6698) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6687..6703 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /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(6711..6741) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6756..6777 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 1021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 1081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 1141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 1201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 1261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 1321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 1381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 1441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 1501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 1561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 1621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 1681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 1741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 1801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 1861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 1921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 1981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 2041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 2101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 2161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 2221 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata 2281 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 2341 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag 2401 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac 2461 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg 2521 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg 2581 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat 2641 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt 2701 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc 2761 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta 2821 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag 2881 cttagccacc atggactaca aagacgatga cgataaagca agggatccaa tgcagagtaa 2941 taaaactttt aacttggaga agcaaaacca tactccaaga aagcatcatc aacatcacca 3001 ccagcagcag caccaccagc agcaacagca gcagccgcca ccaccgccaa tacctgcaaa 3061 tgggcaacag gccagcagcc aaaatgaagg cttgactatt gacctgaaga attttagaaa 3121 accaggagag aagaccttca cccaacgaag ccgtcttttt gtgggaaatc ttcctcccga 3181 catcactgag gaagaaatga ggaaactatt tgagaaatat ggaaaggcag gcgaagtctt 3241 cattcataag gataaaggat ttggctttat ccgcttggaa acccgaaccc tagcggagat 3301 tgccaaagtg gagctggaca atatgccact ccgtggaaag cagctgcgtg tgcgctttgc 3361 ctgccatagt gcatccctta cagttcgaaa ccttcctcag tatgtgtcca acgaactgct 3421 ggaagaagcc ttttctgtgt ttggccaggt agagagggct gtagtcattg tggatgatcg 3481 aggaaggccc tcaggaaaag gcattgttga gttctcaggg aagccagctg ctcggaaagc 3541 tctggacaga tgcagtgaag gctccttcct gctaaccaca tttcctcgtc ctgtgactgt 3601 ggagcccatg gaccagttag atgatgaaga gggacttcca gagaagctgg ttataaaaaa 3661 ccagcaattt cacaaggaac gagagcagcc acccagattt gcacagcctg gctcctttga 3721 gtatgaatat gccatgcgct ggaaggcact cattgagatg gagaagcagc agcaggacca 3781 agtggaccgc aacatcaagg aggctcgtga gaagctggag atggagatgg aagctgcacg 3841 ccatgagcac caggtcatgc taatgagaca ggatttgatg aggcgccaag aagaacttcg 3901 gaggatggaa gagctgcaca accaagaggt gcaaaaacga aagcaactgg agctcaggca 3961 ggaggaagag cgcaggcgcc gtgaagaaga gatgcggcgg cagcaagaag aaatgatgcg 4021 gcgacagcag gaaggattca agggaacctt ccctgatgcg agagagcagg agattcggat 4081 gggtcagatg gctatgggag gtgctatggg cataaacaac agaggtgcca tgccccctgc 4141 tcctgtgcca gctggtaccc cagctcctcc aggacctgcc actatgatgc cggatggaac 4201 tttgggattg accccaccaa caactgaacg ctttggtcag gctgctacaa tggaaggaat 4261 tggggcaatt ggtggaactc ctcctgcatt caaccgtgca gctcctggag ctgaatttgt 4321 cccaaacaaa cgtcgccgat actaactcga gcatgcatct agagggccct attctatagt 4381 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat 4441 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc 4501 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg 4561 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg 4621 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt 4681 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg 4741 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc 4801 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc 4861 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta 4921 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta 4981 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg 5041 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa 5101 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 5161 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 5221 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 5281 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 5341 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc 5401 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa 5461 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt 5521 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 5581 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 5641 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 5701 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 5761 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 5821 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 5881 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 5941 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 6001 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc 6061 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 6121 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 6181 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 6241 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 6301 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc 6361 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 6421 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 6481 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 6541 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 6601 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca 6661 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga 6721 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt 6781 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct g //