LOCUS Exported 9434 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Cre in mammalian cells. ACCESSION . VERSION . KEYWORDS pLenti-hSynapsin-Cre-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9434) AUTHORS Sakurai K, Zhao S, Takatoh J, Rodriguez E, Lu J, Leavitt AD, Fu M, Han BX, Wang F TITLE Capturing and Manipulating Activated Neuronal Ensembles with CANE Delineates a Hypothalamic Social-Fear Circuit. JOURNAL Neuron. 2016 Nov 23;92(4):739-753. doi: 10.1016/j.neuron.2016.10.015. Epub 2016 Oct 27. PUBMED 27974160 REFERENCE 2 (bases 1 to 9434) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9434 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1680..1913 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2098..2142 /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 2440..2557 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2632..3079 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" misc_feature 4126..4714 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4179..4199) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4597..4608) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4717..4733) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4718..4734) /label=pBluescriptKS /note="For pBluescript vector" LTR 5239..5419 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(5445..5462) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5451..5675 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5721..6149 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5808..5827) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6018..6039 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6158..6178) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6163..6492 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6343..6478 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6405..6424 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 6540..6587 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 6606..6980 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 7110..7231 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7147..7166) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7201..7220 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7280..7296) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7280..7296) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7293..7315) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7304..7320 /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(7328..7358) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7373..7394 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7511..7528) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7682..8270) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7762..7781) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8441..9301) /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 9064..9083 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9302..9406) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagcacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2581 agatccagtt tggttaatta agtgtctaga ctgcagaggg ccctgcgtat gagtgcaagt 2641 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc 2701 actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag 2761 gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc 2821 ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc 2881 actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc 2941 cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct 3001 gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt 3061 gtcgtgcctg agagcgcagt cgagaaaccg gctagaggat ccttcgaaac cggtgccacc 3121 atgacgagtg atgaggttcg caagaacctg atggacatgt tcagggatcg ccaggcgttt 3181 tctgagcata cctggaaaat gcttctgtcc gtttgccggt cgtgggcggc atggtgcaag 3241 ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg ttcgcgatta tcttctatat 3301 cttcaggcgc gcggtctggc agtaaaaact atccagcaac atttgggcca gctaaacatg 3361 cttcatcgtc ggtccgggct gccacgacca agtgacagca atgctgtttc actggttatg 3421 cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg caaaacaggc tctagcgttc 3481 gaacgcactg atttcgacca ggttcgttca ctcatggaaa atagcgatcg ctgccaggat 3541 atacgtaatc tggcatttct ggggattgct tataacaccc tgttacgtat agccgaaatt 3601 gccaggatca gggttaaaga tatctcacgt actgacggtg ggagaatgtt aatccatatt 3661 ggcagaacga aaacgctggt tagcaccgca ggtgtagaga aggcacttag cctgggggta 3721 actaaactgg tcgagcgatg gatttccgtc tctggtgtag ctgatgatcc gaataactac 3781 ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat ctgccaccag ccagctatca 3841 actcgcgccc tggaagggat ttttgaagca actcatcgat tgatttacgg cgctaaggat 3901 gactctggtc agagatacct ggcctggtct ggacacagtg cccgtgtcgg agccgcgcga 3961 gatatggccc gcgctggagt ttcaataccg gagatcatgc aagctggtgg ctggaccaat 4021 gtaaatattg tcatgaacta tatccgtaac ctggatagtg aaacaggggc aatggtgcgc 4081 ctgctggaag atggcgatta ggaattcgat atcaagctta tcgataatca acctctggat 4141 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt 4201 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc 4261 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg 4321 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc 4381 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa 4441 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat 4501 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc 4561 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt 4621 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag 4681 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta 4741 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg 4801 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga 4861 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg 4921 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca 4981 caaggctact tccctgattg gcagaactac acaccagggc cagggatcag atatccactg 5041 acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat 5101 gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag 5161 agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag 5221 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct 5281 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta 5341 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca 5401 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc 5461 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc 5521 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg 5581 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa 5641 tagcaggcat gctggggatg cggcgggctc tatggcttct gaggcggaaa gaaccagctg 5701 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt 5761 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt 5821 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct 5881 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg 5941 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga 6001 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc 6061 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga 6121 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt 6181 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 6241 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 6301 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 6361 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 6421 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 6481 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg 6541 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact 6601 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag 6661 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg 6721 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc 6781 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt 6841 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg 6901 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg 6961 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg 7021 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 7081 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 7141 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 7201 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 7261 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 7321 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 7381 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 7441 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 7501 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 7561 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 7621 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 7681 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 7741 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 7801 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 7861 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 7921 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 7981 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 8041 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 8101 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 8161 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 8221 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 8281 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 8341 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 8401 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 8461 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 8521 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 8581 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 8641 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 8701 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 8761 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 8821 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 8881 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 8941 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 9001 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 9061 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 9121 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 9181 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 9241 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 9301 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 9361 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 9421 aagtgccacc tgac //