LOCUS Exported 5673 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses AtPIF6 (Arabidopsis thaliana) preceded by restriction sites. ACCESSION . VERSION . KEYWORDS pOVC1_X_AtPIF6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5673) 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 5673) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5673 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 153..532 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 533..736 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 687..707 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 781..800 /label=T7 /note="T7 promoter, forward primer" promoter 781..799 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1182..1199) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1188..1412 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1458..1886 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1545..1564) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1755..1776 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1895..1915) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1900..2229 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2080..2215 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2142..2161 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2296..3090 /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(2350..2369) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2960..2979 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3264..3385 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3301..3320) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3355..3374 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3434..3450) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3434..3450) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3447..3469) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3458..3474 /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(3482..3512) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3527..3548 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3665..3682) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3836..4424) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3916..3935) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4595..5455) /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 5218..5237 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5456..5560) /gene="bla" /label=AmpR promoter primer_bind complement(5635..5654) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac 61 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct 121 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata 181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact 241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc 421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg 481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 541 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 661 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt 721 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat 781 taatacgact cactataggg agacccaagc tgaccatggc caccggtgaa ttctcaggct 841 ctggatcccc cggtatgatg ttcttaccaa ccgattattg ttgcaggtta agcgatcaag 901 agtatatgga gcttgtgttt gagaatggcc agattcttgc aaagggccaa agatccaacg 961 tttctctgca taatcaacgt accaaatcga tcatggattt gtatgaggca gagtataacg 1021 aggatttcat gaagagtatc atccatggtg gtggtggtgc catcacaaat ctcggggaca 1081 cgcaggttgt tccacaaagt catgttgctg ctgcccatga aacaaacatg ttggaaagca 1141 ataaacatgt tgaccccggg taagtttaaa ccgctgatca gcctcgactg tgccttctag 1201 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 1261 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 1321 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 1381 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 1441 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 1501 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 1561 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 1621 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 1681 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 1741 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 1801 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 1861 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 1921 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 1981 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 2041 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 2101 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 2161 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 2221 ttttgcaaaa agctctcggg agcttgtata tccattttcg gatctgatca agagacagga 2281 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg 2341 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc 2401 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt 2461 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt 2521 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc 2581 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc 2641 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac 2701 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag 2761 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag 2821 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat 2881 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg 2941 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa 3001 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc 3061 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc 3121 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 3181 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 3241 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 3301 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 3361 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 3421 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 3481 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 3541 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 3601 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 3661 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 3721 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 3781 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 3841 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 3901 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 3961 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 4021 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 4081 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 4141 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 4201 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 4261 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 4321 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 4381 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 4441 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 4501 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 4561 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 4621 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 4681 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 4741 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 4801 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 4861 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 4921 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 4981 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 5041 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 5101 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 5161 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 5221 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 5281 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 5341 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 5401 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 5461 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 5521 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 5581 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat 5641 ctgctctgat gccgcatagt taagccagta tct //