LOCUS Exported 5805 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses AsPT1-LOV2-pep (Avena sativa) preceded by restriction sites. ACCESSION . VERSION . KEYWORDS pOVC1_X_AsPT1-LOV2-pep SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5805) AUTHORS Tichy AM, Gerrard EJ, Legrand JMD, Hobbs RM, Janovjak H TITLE Engineering Strategy and Vector Library for the Rapid Generation of Modular Light-Controlled Protein-Protein Interactions. JOURNAL J Mol Biol. 2019 May 29. pii: S0022-2836(19)30317-1. doi: 10.1016/j.jmb.2019.05.033. PUBMED 31150735 REFERENCE 2 (bases 1 to 5805) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5805 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 106..485 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 486..689 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 640..660 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 734..753 /label=T7 /note="T7 promoter, forward primer" promoter 734..752 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1267..1284) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1273..1497 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1543..1971 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1630..1649) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1840..1861 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1980..2000) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1985..2314 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2165..2300 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2227..2246 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2381..3175 /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(2435..2454) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3045..3064 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3349..3470 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3386..3405) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3440..3459 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3519..3535) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3519..3535) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3532..3554) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3543..3559 /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(3567..3597) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3612..3633 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3750..3767) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3921..4509) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4001..4020) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4680..5540) /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 5303..5322 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5541..5645) /gene="bla" /label=AmpR promoter primer_bind complement(5720..5739) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta 61 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga 121 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc 181 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 241 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 301 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 361 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 421 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 481 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg 541 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac 601 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg 661 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact 721 ggcttatcga aattaatacg actcactata gggagaccca agctgaccat ggccaccggt 781 gaattctcag gctctggatc ccccgggttg gctgctgcac ttgaacgtat tgagaagaac 841 tttgtcatta ctgacccaag attgccagat aatcccatta tattcgcgtc cgatagtttc 901 ttgcagttga cagaatatag ccgtgaagaa attttgggaa gaaactgcag gtttctacaa 961 ggtcctgaaa ctgatcgcgc gacagtgaga aaaattagag atgccataga taaccaaaca 1021 gaggtcactg ttcagctgat taattataca aagagtggta aaaagttctg gaacctcttt 1081 cacttgcagc ctatgcgaga tcagaaggga gatgtccagt actttattgg ggttcagttg 1141 gatggaactg agcatgtccg agatgctgcc gagagagagg cagtcatgct ggcgaagaaa 1201 actgcagaag agattgataa agcggtggat acctgggtgc ccgggtaagt ttaaaccgct 1261 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc 1321 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg 1381 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca 1441 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt 1501 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg 1561 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc 1621 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc 1681 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg 1741 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg 1801 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg 1861 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt 1921 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg 1981 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 2041 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 2101 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 2161 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 2221 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg 2281 aggaggcttt tttggaggcc taggcttttg caaaaagctc tcgggagctt gtatatccat 2341 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt 2401 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 2461 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 2521 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct 2581 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 2641 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 2701 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 2761 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 2821 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 2881 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac 2941 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 3001 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 3061 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 3121 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg 3181 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 3241 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 3301 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt 3361 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 3421 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt 3481 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga 3541 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 3601 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 3661 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 3721 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 3781 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 3841 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 3901 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 3961 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 4021 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 4081 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 4141 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 4201 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 4261 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 4321 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 4381 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 4441 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 4501 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 4561 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 4621 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 4681 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 4741 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 4801 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 4861 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 4921 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 4981 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 5041 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 5101 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 5161 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 5221 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 5281 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 5341 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 5401 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 5461 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 5521 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 5581 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 5641 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc 5701 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc 5761 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaa //