LOCUS Exported 7687 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Lentiviral vector that expresses GFP and an shRNA targeting Esr1 (in pLL3.7).. ACCESSION . VERSION . KEYWORDS pLV-shEsr1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7687) AUTHORS Satta R, Certa B, He D, Lasek AW TITLE Estrogen Receptor beta in the Nucleus Accumbens Regulates the Rewarding Properties of Cocaine in Female Mice. JOURNAL Int J Neuropsychopharmacol. 2018 Apr 1;21(4):382-392. doi: 10.1093/ijnp/pyx118. PUBMED 29294029 REFERENCE 2 (bases 1 to 7687) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7687 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 618..816 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 834..1014 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1061..1186 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1683..1916 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2101..2145 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2443..2560 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" primer_bind 2850..2870 /label=mU6-F /note="Mouse U6 promoter, forward primer" primer_bind 2988..3004 /label=KS primer /note="common sequencing primer, one of multiple similar variants" protein_bind 3046..3079 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." enhancer 3195..3498 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3499..3702 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" regulatory 3741..3750 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3747..4463 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3792..3813) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4053..4072) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4400..4421 /label=EGFP-C /note="EGFP, forward primer" protein_bind 4486..4519 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 4575..5163 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4628..4648) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5046..5057) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5166..5182) /label=KS primer /note="common sequencing primer, one of multiple similar variants" LTR 5692..5872 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(5934..6522) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6014..6033) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6693..7553) /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 7316..7335 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7554..7658) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc 841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta 901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact 961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg 1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg 1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat 1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg 1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa 1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct 1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac 1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca 1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa 1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat 1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta 1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga 1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca 1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat 1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag 1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg 1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg 1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa 2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa 2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac 2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt 2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca 2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa 2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg 2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg 2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac 2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag 2581 cagagatcca gtttggttag taccgggccc gctctagaga tccgacgcgc catctctagg 2641 cccgcgccgg ccccctcgca cagacttgtg ggagaagctc ggctactccc ctgccccggt 2701 taatttgcat ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat 2761 aatctgttct ttttaatact agctacattt tacatgatag gcttggattt ctataagaga 2821 tacaaatact aaattattat tttaaaaaac agcacaaaag gaaactcacc ctaactgtaa 2881 agtaattgtg tgttttgaga ctataaatat cccttggaga aaagccttgt ttggcatgga 2941 gcatctctac attcaagaga tgtagagatg ctccatgcct tttttctcga ggtcgacggt 3001 atcgataagc tcgcttcacg agattccagc aggtcgaggg acctaataac ttcgtatagc 3061 atacattata cgaagttata ttaagggttc caagcttaag cggccgcgtg gataaccgta 3121 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca 3181 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac 3241 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact 3301 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa 3361 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg 3421 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta 3481 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg 3541 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg 3601 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg 3661 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag 3721 atccgctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg 3781 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg 3841 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg 3901 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct 3961 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 4021 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg 4081 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 4141 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct 4201 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca 4261 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 4321 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc 4381 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc 4441 tcggcatgga cgagctgtac aagtaggaat tcgtcgaggg acctaataac ttcgtatagc 4501 atacattata cgaagttata catgtttaag ggttccggtt ccactaggta caattcgata 4561 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc 4621 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg 4681 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc 4741 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg 4801 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg 4861 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga 4921 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct 4981 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg 5041 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc 5101 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc 5161 cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc 5221 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg 5281 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat 5341 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga 5401 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac 5461 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta 5521 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac 5581 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac 5641 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc 5701 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag 5761 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct 5821 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga 5881 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 5941 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6001 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 6061 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 6121 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 6181 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 6241 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 6301 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 6361 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 6421 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 6481 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6541 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6601 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6661 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 6721 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 6781 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 6841 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 6901 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 6961 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 7021 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 7081 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 7141 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 7201 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 7261 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 7321 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 7381 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 7441 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 7501 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7561 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7621 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7681 cctgacg //