LOCUS Exported 9958 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Constitutive expression of hICN1, e.g. for hedgehog-induction. Based on addgene #17623, with CMV.GFP removed.. ACCESSION . VERSION . KEYWORDS EF.hICN1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9958) AUTHORS Maier J, Elmenofi S, Taschauer A, Anton M, Sami H, Ogris M TITLE Luminescent and fluorescent triple reporter plasmid constructs for Wnt, Hedgehog and Notch pathway. JOURNAL PLoS One. 2019 Dec 20;14(12):e0226570. doi: 10.1371/journal.pone.0226570. eCollection 2019. PUBMED 31860685 REFERENCE 2 (bases 1 to 9958) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9958 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..1179 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 232..1170 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1127..1147 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" promoter 1196..1214 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" LTR 3778..4011 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 4083..4204 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4120..4139) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4174..4193 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 4244..4379 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4306..4325 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter complement(4400..4418) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4428..4445) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4428..4444) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4437..4459) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 4586..5041 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4673..4692) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4883..4904 /label=F1ori-F /note="F1 origin, forward primer" promoter 5067..5171 /gene="bla" /label=AmpR promoter CDS 5172..6032 /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(5390..5409) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6203..6791 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6692..6711 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6945..6962 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7079..7100 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7115..7145 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7153..7169 /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 7158..7180 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7177..7193 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7177..7193 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 7212..7232 /label=T3 /note="T3 promoter, forward primer" promoter 7214..7232 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 7260..7486 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 7487..7667 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 7714..7839 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 9109..9342 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 9527..9571 /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" ORIGIN 1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc 241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt 301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg 361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg 421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct 481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg 541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc 601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga 661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct 721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca 781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg 841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg 901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat 961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg 1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa 1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca 1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata 1201 cgactcacta tagggagacc caagctggct aggtaagctt gatgatccgg aggcatgcgg 1261 cggcagcatg gccagctctg gttccctgag ggcttcaaag tgtctgaggc cagcaagaag 1321 aagcggcggg agcccctcgg cgaggactcc gtgggcctca agcccctgaa gaacgcttca 1381 gacggtgccc tcatggacga caaccagaat gagtgggggg acgaggacct ggagaccaag 1441 aagttccggt tcgaggagcc cgtggttctg cctgacctgg acgaccagac agaccaccgg 1501 cagtggactc agcagcacct ggatgccgct gacctgcgca tgtctgccat ggcccccaca 1561 ccgccccagg gtgaggttga cgccgactgc atggacgtca atgtccgcgg gcctgatggc 1621 ttcaccccgc tcatgatcgc ctcctgcagc gggggcggcc tggagacggg caacagcgag 1681 gaagaggagg acgcgccggc cgtcatctcc gacttcatct accagggcgc cagcctgcac 1741 aaccagacag accgcacggg cgagaccgcc ttgcacctgg ccgcccgcta ctcacgctct 1801 gattccgcca agcgcctgct ggaggccagc gcagatgcca acatccagga caacatgggc 1861 cgcaccccgc tgcatgcggc tgtgtctgcc gacgcacaag gtgtcttcca gatcctgatc 1921 cggaaccgag ccacagacct ggatgcccgc atgcatgatg gcacgacgcc actgatcctg 1981 gctgcccgcc tggccgtgga gggcatgctg gaggacctca tcaactcaca cgccgacgtc 2041 aacgccgtag atgacctggg caagtccgcc ctgcactggg ccgccgccgt gaacaatgtg 2101 gatgccgcag ttgtgctcct gaagaacggg gctaacaaag atatgcagaa caacagggag 2161 gagacacccc tgtttctggc cgcccgggag ggcagctacg agaccgccaa ggtgctgctg 2221 gaccactttg ccaaccggga catcacggat catatggacc gcctgccgcg cgacatcgca 2281 caggagcgca tgcatcacga catcgtgagg ctgctggacg agtacaacct ggtgcgcagc 2341 ccgcagctgc acggagcccc gctggggggc acgcccaccc tgtcgccccc gctctgctcg 2401 cccaacggct acctgggcag cctcaagccc ggcgtgcagg gcaagaaggt ccgcaagccc 2461 agcagcaaag gcctggcctg tggaagcaag gaggccaagg acctcaaggc acggaggaag 2521 aagtcccagg atggcaaggg ctgcctgctg gacagctccg gcatgctctc gcccgtggac 2581 tccctggagt caccccatgg ctacctgtca gacgtggcct cgccgccact gctgccctcc 2641 ccgttccagc agtctccgtc cgtgcccctc aaccacctgc ctgggatgcc cgacacccac 2701 ctgggcatcg ggcacctgaa cgtggcggcc aagcccgaga tggcggcgct gggtgggggc 2761 ggccggctgg cctttgagac tggcccacct cgtctctccc acctgcctgt ggcctctggc 2821 accagcaccg tcctgggctc cagcagcgga ggggccctga atttcactgt gggcgggtcc 2881 accagtttga atggtcaatg cgagtggctg tcccggctgc agagcggcat ggtgccgaac 2941 caatacaacc ctctgcgggg gagtgtggca ccaggccccc tgagcacaca ggccccctcc 3001 ctgcagcatg gcatggtagg cccgctgcac agtagccttg ctgccagcgc cctgtcccag 3061 atgatgagct accagggcct gcccagcacc cggctggcca cccagcctca cctggtgcag 3121 acccagcagg tgcagccaca aaacttacag atgcagcagc agaacctgca gccagcaaac 3181 atccagcagc agcaaagcct gcagccgcca ccaccaccac cacagccgca ccttggcgtg 3241 agctcagcag ccagcggcca cctgggccgg agcttcctga gtggagagcc gagccaggca 3301 gacgtgcagc cactgggccc cagcagcctg gcggtgcaca ctattctgcc ccaggagagc 3361 cccgccctgc ccacgtcgct gccatcctcg ctggtcccac ccgtgaccgc agcccagttc 3421 ctgacgcccc cctcgcagca cagctactcc tcgcctgtgg acaacacccc cagccaccag 3481 ctacaggtgc ctgagcaccc cttcctcacc ccgtcccctg agtcccctga ccagtggtcc 3541 agctcgtccc cgcattccaa cgtctccgac tggtccgagg gcgtctccag ccctcccacc 3601 agcatgcagt cccagatcgc ccgcattccg gaggccttca agtaaacggc gcgtcagatc 3661 cactagtaac ggccgccagt gtgctggaat tatcgaattc gagctcggta cctttaagac 3721 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg 3781 aagggctaat tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct 3841 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc 3901 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg 3961 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt 4021 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga 4081 ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca 4141 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat 4201 cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc ccctaactcc 4261 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 4321 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 4381 agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt 4441 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat 4501 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag 4561 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt 4621 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc 4681 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg 4741 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat 4801 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg 4861 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct 4921 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa 4981 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt 5041 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 5101 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 5161 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 5221 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 5281 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 5341 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 5401 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 5461 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 5521 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 5581 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 5641 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 5701 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 5761 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 5821 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 5881 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 5941 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 6001 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 6061 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 6121 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 6181 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 6241 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 6301 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 6361 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6421 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6481 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6541 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6601 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6661 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6721 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6781 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6841 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6901 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 6961 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 7021 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 7081 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta 7141 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 7201 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat 7261 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc 7321 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg 7381 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc 7441 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt 7501 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca 7561 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa 7621 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac 7681 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct 7741 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact 7801 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt 7861 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa 7921 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa 7981 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca 8041 gaagaactta gatcattata taatacagta gcaaccctct attgtgtggg taatgactcc 8101 aacttactga tagtgtttta tgttcagata atgcccgatg accttgtcat gcaactccac 8161 cgattttgag aacgacagta acttccgtcc cagccttgcc agatgttgtc tcagattcag 8221 attatgtcgc tcaatgcgct gagtgtaacg cttgctgata acgtgcagct ttcccttcag 8281 gcgtgattca tacagcggcc agccatccgt catctatacc acgacctcaa aggccgacag 8341 caggctcaga agacgctcca gtgtggccag agtgcgttca ccgaagacgt gcgccacaac 8401 cgtcctccgt atcctgtcat acgcgtaaaa cagccagcgc tgacgtgatt tagcaccgac 8461 gtagccccac tgttcgtcca tttcagcgca gacaatcaca tcactgcccg gttgtatgcg 8521 cgaggttacc gactgcggcc tgagtttttt aagtgacgta aaaccgtgtt gaggccaacg 8581 cccataatgc gtgcactggc gcgacatccg acgccattca tggccatatc aatgattttc 8641 tggtgcttac cgggctgaga ggcggtgtaa gtgaactgta gttgccatgt tttacggcaa 8701 ggagagcaga gatagcgctg atgtccggca gtgcttttgc cgttacgcac cacgccttca 8761 gtagcggagc aggaaggaca tctgatggaa atggaagcca cgcaagcacc ttaaaatcac 8821 catcatacac taaatcagta agttggcagc attacctatt gtgtgcatca aaggatagag 8881 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc 8941 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg 9001 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac 9061 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt 9121 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt 9181 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat 9241 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag 9301 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc 9361 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct 9421 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac 9481 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga 9541 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct 9601 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt 9661 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac 9721 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga 9781 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc gatcacgaga 9841 ctagcctcga gaagcttgat gacggtatcg ataagctttg caaagatgga taaagtttta 9901 aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag tgggaatt //