LOCUS Exported 7117 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCDNA3.1-nv5-ER beta SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7117) AUTHORS Wittmann BM, Sherk A, McDonnell DP TITLE Definition of functionally important mechanistic differences among selective estrogen receptor down-regulators. JOURNAL Cancer Res. 2007 Oct 1. 67(19):9549-60. PUBMED 17909066 REFERENCE 2 (bases 1 to 7117) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7117 /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" CDS 914..955 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 956..976 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" protein_bind 1007..1031 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(2668..2692) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(2711..2728) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2717..2941 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2987..3415 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3074..3093) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3284..3305 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3424..3444) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3429..3758 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3609..3744 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3671..3690 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3825..4619 /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(3879..3898) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4489..4508 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4793..4914 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4830..4849) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4884..4903 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4963..4979) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4963..4979) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4976..4998) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4987..5003 /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(5011..5041) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5056..5077 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5194..5211) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5365..5950) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5445..5464) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6121..6981) /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 6744..6763 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6982..7086) /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 gctggctagt 901 taagcttacc atgggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacggaaaa 961 cctgtatttt cagggcccgg atccgtcgac gaattctgca gatatcacaa gtttgtacaa 1021 aaaagcaggc tttaaaggaa ccaattcagt cgacgatata aaaaactcac catctagcct 1081 taattctcct tcctcctaca actgcagtca atccatctta cccctggagc acggctccat 1141 atacatacct tcctcctatg tagacagcca ccatgaatat ccagccatga cattctatag 1201 ccctgctgtg atgaattaca gcattcccag caatgtcact aacttggaag gtgggcctgg 1261 tcggcagacc acaagcccaa atgtgttgtg gccaacacct gggcaccttt ctcctttagt 1321 ggtccatcgc cagttatcac atctgtatgc ggaacctcaa aagagtccct ggtgtgaagc 1381 aagatcgcta gaacacacct tacctgtaaa cagagagaca ctgaaaagga aggttagtgg 1441 gaaccgttgc gccagccctg ttactggtcc aggttcaaag agggatgctc acttctgcgc 1501 tgtctgcagc gattacgcat cgggatatca ctatggagtc tggtcgtgtg aaggatgtaa 1561 ggcctttttt aaaagaagca ttcaaggaca taatgattat atttgtccag ctacaaatca 1621 gtgtacaatc gataaaaacc ggcgcaagag ctgccaggcc tgccgacttc ggaagtgtta 1681 cgaagtggga atggtgaagt gtggctcccg gagagagaga tgtgggtacc gccttgtgcg 1741 gagacagaga agtgccgacg agcagctgca ctgtgccggc aaggccaaga gaagtggcgg 1801 ccacgcgccc cgagtgcggg agctgctgct ggacgccctg agccccgagc agctagtgct 1861 caccctcctg gaggctgagc cgccccatgt gctgatcagc cgccccagtg cgcccttcac 1921 cgaggcctcc atgatgatgt ccctgaccaa gttggccgac aaggagttgg tacacatgat 1981 cagctgggcc aagaagattc ccggctttgt ggagctcagc ctgttcgacc aagtgcggct 2041 cttggagagc tgttggatgg aggtgttaat gatggggctg atgtggcgct caattgacca 2101 ccccggcaag ctcatctttg ctccagatct tgttctggac agggatgagg ggaaatgcgt 2161 agaaggaatt ctggaaatct ttgacatgct cctggcaact acttcaaggt ttcgagagtt 2221 aaaactccaa cacaaagaat atctctgtgt caaggccatg atcctgctca attccagtat 2281 gtaccctctg gtcacagcga cccaggatgc tgacagcagc cggaagctgg ctcacttgct 2341 gaacgccgtg accgatgctt tggtttgggt gattgccaag agcggcatct cctcccagca 2401 gcaatccatg cgcctggcta acctcctgat gctcctgtcc cacgtcaggc atgcgagtaa 2461 caagggcatg gaacatctgc tcaacatgaa gtgcaaaaat gtggtcccag tgtatgacct 2521 gctgctggag atgctgaatg cccacgtgct tcgcgggtgc aagtcctcca tcacggggtc 2581 cgagtgcagc ccggcagagg acagtaaaag caaagagggc tcccagaacc cacagtctca 2641 gtgagcggcc gcactcgaga tatctagacc cagctttctt gtacaaagtg gtgataaacc 2701 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2761 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2821 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2881 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2941 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 3001 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 3061 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 3121 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 3181 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 3241 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 3301 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 3361 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 3421 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3481 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3601 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3661 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3721 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3781 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3841 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3901 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3961 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 4021 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 4081 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 4141 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 4201 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4261 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4321 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 4381 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 4441 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4501 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4561 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4621 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 4681 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4741 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 4801 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4861 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4921 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 4981 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 5041 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 5101 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 5161 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 5221 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5281 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5341 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 5401 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5461 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5521 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 5581 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5641 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5701 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5761 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 5821 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 5881 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5941 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 6001 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 6061 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 6121 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 6181 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6241 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6301 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 6361 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6421 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6481 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6541 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6601 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6661 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6721 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6781 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6841 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6901 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6961 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 7021 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 7081 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //