LOCUS Exported 5790 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS T7-Elongin C-pcDNA3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5790) AUTHORS Lonergan KM, Iliopoulos O, Ohh M, Kamura T, Conaway RC, Conaway JW, Kaelin WG TITLE Regulation of hypoxia-inducible mRNAs by the von Hippel-Lindau tumor suppressor protein requires binding to complexes containing elongins B/C and Cul2. JOURNAL Mol Cell Biol. 1998 Feb . 18(2):732-41. PUBMED 9447969 REFERENCE 2 (bases 1 to 5790) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5790 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 895..904 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(1342..1360) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1343..1360) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1380..1397) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1386..1610 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1656..2084 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1743..1762) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1953..1974 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2093..2113) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2098..2428 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2279..2414 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2341..2360 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2495..3289 /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(2549..2568) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3159..3178 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3463..3584 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3500..3519) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3554..3573 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3633..3649) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3633..3649) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3646..3668) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3657..3673 /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(3681..3711) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3726..3747 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3864..3881) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4035..4623) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4115..4134) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4794..5654) /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 5417..5436 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5655..5759) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc 901 atggctagca tgactggtgg acagcaaatg ggatccatgg atggagagga gaaaacctat 961 ggtggctgtg aaggacctga tgccatgtat gtcaaattga tatcatctga tggccatgaa 1021 tttattgtaa aaagagaaca tgcattaaca tcaggcacga taaaagccat gttgagtggc 1081 ccaggtcagt ttgctgagaa cgaaaccaat gaggtcaatt ttagagagat accttcacat 1141 gtgctatcga aagtatgcat gtattttacg tacaaggttc gctacactaa cagctccacc 1201 gagattcctg aattcccaat tgcacctgaa attgcactgg aactgctgat ggctgcgaac 1261 ttcttagatt gttaaggtct ccaattctgc agatatccat cacactggcg gccgctcgag 1321 catgcatcta gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc 1381 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 1441 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 1501 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 1561 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 1621 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag 1681 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc 1741 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc 1801 tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa 1861 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg 1921 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac 1981 actcaaccct atctcggtct attcttttga tttataaggg attttgggga tttcggccta 2041 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg 2101 tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat 2161 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 2221 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 2281 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 2341 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 2401 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 2461 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 2521 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 2581 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 2641 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 2701 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 2761 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 2821 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 2881 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 2941 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 3001 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 3061 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 3121 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 3181 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 3241 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 3301 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 3361 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 3421 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 3481 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 3541 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 3601 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 3661 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 3721 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 3781 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 3841 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 3901 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 3961 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 4021 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 4081 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 4141 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4201 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 4261 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4321 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4381 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4441 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 4501 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4561 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 4621 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 4681 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 4741 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 4801 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 4861 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 4921 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 4981 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 5041 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 5101 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 5161 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 5221 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 5281 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 5341 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 5401 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 5461 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 5521 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 5581 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 5641 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 5701 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 5761 acatttcccc gaaaagtgcc acctgacgtc //