LOCUS Exported 7492 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express an NT-3-BDNF hybrid neurotrophic factor in mammalian cells. ACCESSION . VERSION . KEYWORDS pLV-NT-3/D15A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7492) AUTHORS Enomoto M, Bunge MB, Tsoulfas P TITLE A multifunctional neurotrophin with reduced affinity to p75NTR enhances transplanted Schwann cell survival and axon growth after spinal cord injury. JOURNAL Exp Neurol. 2013 Oct;248:170-82. doi: 10.1016/j.expneurol.2013.06.013. Epub 2013 Jun 20. PUBMED 23792206 REFERENCE 2 (bases 1 to 7492) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7492 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 21..125 /gene="bla" /label=AmpR promoter CDS 126..986 /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 complement(344..363) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1157..1745 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1646..1665 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1899..1916 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2033..2054 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2069..2099 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2107..2123 /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)." primer_bind 2112..2134 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2131..2147 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2131..2147 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2166..2186 /label=T3 /note="T3 promoter, forward primer" promoter 2168..2186 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 2214..2440 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 2441..2621 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 2668..2793 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3286..3519 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 3704..3748 /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 4015..4131 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 4154..4457 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4458..4661 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4612..4632 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 4658..4682 /label=LNCX /note="Human CMV promoter, forward primer" misc_feature 5549..6137 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5602..5622) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6020..6031) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6224..6457 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6529..6650 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6566..6585) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6620..6639 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6690..6825 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6752..6771 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(6845..6864) /label=T7 /note="T7 promoter, forward primer" promoter complement(6846..6864) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6874..6891) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6874..6890) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6883..6905) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 7032..7487 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7119..7138) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7329..7350 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc 721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc 781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga 1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt 1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag 1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca 1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga 2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt 2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca 2161 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc 2221 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca 2281 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt 2341 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg 2401 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca 2461 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 2521 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 2581 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac 2641 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg 2701 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga 2761 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg 2821 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata 2881 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca 2941 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa 3001 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata 3061 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc 3121 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag 3181 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa 3241 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct 3301 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca 3361 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga 3421 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat 3481 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg 3541 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga 3601 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag 3661 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt 3721 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg 3781 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc 3841 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca 3901 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag 3961 agacagagac agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa 4021 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac 4081 agacatacaa actaaagaat tacaaaaaca aattacaaaa ttcaaaattt tatcgataag 4141 cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 4201 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 4261 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 4321 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 4381 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 4441 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 4501 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 4561 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 4621 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga 4681 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga ctctagagga 4741 tcctacaggt gaacaagacc atgtccatct tgttttatgt gatatttctc gcttatctcc 4801 gtggcatcca aggtaacaac atggatcaaa ggagtttgcc agaagactcg ctcaattccc 4861 tcattattaa gctgatccag gcagatattt tgaaaaacaa gctctccaag cagatggtgg 4921 acgttaagga aaattaccag agcaccctgc ccaaagctga ggctccccga gagccggagc 4981 ggggagggcc cgccaagtca gcattccagc cggtgattgc aatggacacc gaactgctgc 5041 gacaacagag acgctacaac tcaccgcggg tcctgctgag cgacagcacc cccttggagc 5101 ccccgccctt gtatctcatg gaggattacg tgggcagccc cgtggtggcg aacagaacat 5161 cacggcggaa acggtacgcg gagcataaga gtcaccgagg ggagtactcg gtatgtgcca 5221 gtgaaagctt gtgggtgacc gacaagtcat cggccatcga cattcgggga caccaggtca 5281 cggtgctggg ggagatcaaa acgggcaact ctcccgtcaa acaatatttt tatgaaacgc 5341 gatgtaagga agccaggccg gtcaaaaacg gttgcagggg tattgatgat aaacactgga 5401 actctcagtg caaaacatcc caaacctacg tccgagcact gacttcagag aacaataaac 5461 tcgtgggctg gcggtggata cggatagaca cgtcctgtgt gtgtgccttg tcgagaaaaa 5521 tcggaagaac atgaattggc atctcgacaa tcaacctctg gattacaaaa tttgtgaaag 5581 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat 5641 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc 5701 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg 5761 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct 5821 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct 5881 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg 5941 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac 6001 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct 6061 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct 6121 ttgggccgcc tccccgcctg gaattcgagc tcggtacctt taagaccaat gacttacaag 6181 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac 6241 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc 6301 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg 6361 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc 6421 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt 6481 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt 6541 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 6601 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg 6661 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 6721 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg 6781 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa 6841 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga 6901 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag 6961 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa 7021 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 7081 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 7141 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 7201 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 7261 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 7321 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 7381 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 7441 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tg //