LOCUS Exported 7192 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses HA-tagged ER wild-type in mammalian cells . ACCESSION . VERSION . KEYWORDS pcDNA-HA-ER WT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7192) TITLE pcDNA-HA-ER WT JOURNAL Unpublished REFERENCE 2 (bases 1 to 7192) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7192 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 143..522 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 523..726 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 677..697 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 771..790 /label=T7 /note="T7 promoter, forward primer" promoter 771..789 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 852..878 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1770..2663 /codon_start=1 /gene="ESR1" /product="estrogen nuclear receptor alpha ligand-binding domain" /label=ER-LBD /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL QKYYITGEAEGFPATV" primer_bind complement(2691..2708) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2697..2921 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2967..3395 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3054..3073) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3264..3285 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3404..3424) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3409..3738 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3589..3724 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3651..3670 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3805..4599 /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(3859..3878) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4469..4488 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4773..4894 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4810..4829) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4864..4883 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4943..4959) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4943..4959) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4956..4978) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4967..4983 /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(4991..5021) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5036..5057 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5174..5191) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5345..5933) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5425..5444) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6104..6964) /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 6727..6746 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6965..7069) /gene="bla" /label=AmpR promoter primer_bind complement(7144..7163) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc 61 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat 121 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt 181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat 241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt 301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa 361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc 421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct 481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag 541 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt 601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac 661 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc 721 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact 781 cactataggg agacccaagc tggctagcgt ttaaacttaa gcttggtacc gagctcggat 841 ccgccaccat gtatccatat gacgtcccag actatgccac catgaccctc cacaccaaag 901 catctgggat ggccctactg catcagatcc aagggaacga gctggagccc ctgaaccgtc 961 cgcagctcaa gatccccctg gagcggcccc tgggcgaggt gtacctggac agcagcaagc 1021 ccgccgtgta caactacccc gagggcgccg cctacgagtt caacgccgcg gccgccgcca 1081 acgcgcaggt ctacggtcag accggcctcc cctacggccc cgggtctgag gctgcggcgt 1141 tcggctccaa cggcctgggg ggtttccccc cactcaacag cgtgtctccg agcccgctga 1201 tgctactgca cccgccgccg cagctgtcgc ctttcctgca gccccacggc cagcaggtgc 1261 cctactacct ggagaacgag cccagcggct acacggtgcg cgaggccggc ccgccggcat 1321 tctacaggcc aaattcagat aatcgacgcc agggtggcag agaaagattg gccagtacca 1381 atgacaaggg aagtatggct atggaatctg ccaaggagac tcgctactgt gcagtgtgca 1441 atgactatgc ttcaggctac cattatggag tctggtcctg tgagggctgc aaggccttct 1501 tcaagagaag tattcaagga cataacgact atatgtgtcc agccaccaac cagtgcacca 1561 ttgataaaaa caggaggaag agctgccagg cctgccggct ccgcaaatgc tacgaagtgg 1621 gaatgatgaa aggtgggata cgaaaagacc gaagaggagg gagaatgttg aaacacaagc 1681 gccagagaga tgatggggag ggcaggggtg aagtggggtc tgctggagac atgagagctg 1741 ccaacctttg gccaagcccg ctcatgatca aacgctctaa gaagaacagc ctggccttgt 1801 ccctgacggc cgaccagatg gtcagtgcct tgttggatgc tgagcccccc atactctatt 1861 ccgagtatga tcctaccaga cccttcagtg aagcttcgat gatgggctta ctgaccaacc 1921 tggcagacag ggagctggtt cacatgatca actgggcgaa gagggtgcca ggctttgtgg 1981 atttgaccct ccatgatcag gtccaccttc tagaatgtgc ctggctagag atcctgatga 2041 ttggtctcgt ctggcgctcc atggagcacc cagggaagct actgtttgct cctaacttgc 2101 tcttggacag gaaccaggga aaatgtgtag agggcatggt ggagatcttc gacatgctgc 2161 tggctacatc atctcggttc cgcatgatga atctgcaggg agaggagttt gtgtgcctca 2221 aatctattat tttgcttaat tctggagtgt acacatttct gtccagcacc ctgaagtctc 2281 tggaagagaa ggaccatatc caccgagtcc tggacaagat cacagacact ttgatccacc 2341 tgatggccaa ggcaggcctg accctgcagc agcagcacca gcggctggcc cagctcctcc 2401 tcatcctctc ccacatcagg cacatgagta acaaaggcat ggagcatctg tacagcatga 2461 agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga gatgctggac gcccaccgcc 2521 tacatgcgcc cactagccgt ggaggggcat ccgtggagga gacggaccaa agccacttgg 2581 ccactgcggg ctctacttca tcgcattcct tgcaaaagta ttacatcacg ggggaggcag 2641 agggtttccc tgccacggtc tgattgggcc cgtttaaacc cgctgatcag cctcgactgt 2701 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga 2761 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag 2821 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga 2881 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac 2941 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg 3001 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt 3061 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg 3121 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga 3181 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac 3241 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc 3301 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa 3361 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta 3421 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 3481 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 3541 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 3601 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 3661 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga 3721 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa 3781 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 3841 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 3901 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 3961 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg 4021 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4081 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4141 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4201 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4261 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4321 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4381 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4441 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4501 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4561 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4621 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4681 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4741 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt 4801 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 4861 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta 4921 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 4981 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag 5041 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 5101 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 5161 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 5221 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 5281 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 5341 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 5401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 5461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 5521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 5581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 5641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 5701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 5761 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 5821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 5881 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 5941 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 6001 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 6061 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 6121 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 6181 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 6241 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 6301 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 6361 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 6421 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 6481 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 6541 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 6601 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 6661 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 6721 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 6781 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 6841 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 6901 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 6961 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 7021 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 7081 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct 7141 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg ct //